Methods of treating ocular diseases using lipid binding protein-based complexes

Abstract

Disclosed herein are methods of treating an ocular disease, such as an ocular disease associated with lipid accumulation, with a lipid-binding protein-based complex, such as CER-001; the present invention relates to lipid binding protein-based complexes, compositions comprising lipid binding protein-based complexes as one or more ophthalmic drug carriers, and uses thereof.

Description

[0001] 1. Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 328,088, filed on April 6, 2022, the contents of which are incorporated herein by reference in their entirety.

[0003] 2. Sequence Listing

[0004] This application contains a sequence listing, which has been submitted electronically and is incorporated herein by reference in its entirety. The electronic copy was created on March 28, 2023, is an XML file named CRN-041B-WO_ST26, and is 3,216 bytes in size.

[0005] 3. Background

[0006] 3.1. Eye diseases

[0007] The vertebrate eye is a complex sensory organ composed of multiple distinct tissues, each with its own unique biochemical composition, structure, and physiological functions. Key among them are the retina, lens, and cornea, which work together to bring photons of light into the eye, focus them correctly on the retina, and convert their energy into electrochemical signals that are transmitted to the brain, where they are ultimately processed into a coherent visual image. Defects in any or all of these tissues, whether congenital or acquired, either through disease processes or through trauma, impair vision and can ultimately lead to complete and irreversible blindness. Lipids and lipid-soluble compounds are essential components of the cells and tissues that make up the eye, and defects in their synthesis, intracellular and extracellular transport, and turnover underlie a variety of important, common, and often severely debilitating eye diseases.

[0008] Eye diseases can have many causes, such as genetics, infection, and aging. Some eye diseases are associated with lipid accumulation in or near the eye, such as ichthyosis, dry eye disease (e.g., associated with meibomian gland dysfunction or tear gland dysfunction), blepharitis, uveitis, corneal diseases such as lipid keratopathy, dry macular degeneration (dry AMD), Stargardt disease, and Leber's idiopathic stellate neuroretinitis.

[0009] 3.2. Lecithin cholesterol acyltransferase

[0010] Phosphatidylcholine cholesterol acyltransferase (LCAT) is an enzyme produced by the liver and is a key enzyme in the reverse cholesterol transport (RCT) pathway. The function of the RCT pathway is to eliminate cholesterol from most extrahepatic tissues and is essential for maintaining the structure and function of most cells in the body. RCT consists of three major steps: (a) cholesterol efflux, which is the initial removal of cholesterol from various peripheral cell pools; (b) cholesterol esterification by the action of phosphatidylcholine: cholesterol acyltransferase (LCAT), which prevents the effluxed cholesterol from reentering the cell; and (c) uptake of high-density lipoprotein (HDL)-cholesterol and cholesterol esters by hepatocytes for hydrolysis, followed by recirculation, storage, excretion in bile, or catabolism to bile acids.

[0011] LCAT circulates in the plasma partially related to HDL.LCAT converts cell-derived cholesterol into cholesterol ester, which is isolated in HDL for removal (referring to Jonas 2000, Biochim.Biophys.Acta 1529 (1-3): 245-56).Cholesterol ester transfer protein (CETP) and phospholipid transfer protein (PLTP) contribute to the further remodeling of circulating HDL colonies.CETP transfers the cholesterol ester produced by LCAT to other lipoproteins, particularly lipoproteins comprising ApoB, such as very low density lipoprotein (VLDL) and low density lipoprotein (LDL).PLTP provides lecithin to HDL.HDL triglycerides are decomposed by extracellular liver triglyceride lipase, and lipoprotein cholesterol is removed by liver via multiple mechanisms.

[0012] LCAT deficiency leads to the accumulation of unesterified cholesterol in certain body tissues. Cholesterol flows out of cells in the form of free cholesterol and is transported in HDL in the form of esterified cholesterol. LCAT is an enzyme that esterifies the free cholesterol on HDL into cholesterol esters and makes HDL mature. LCAT deficiency will not allow HDL to mature, leading to the rapid catabolism of its circulating apoA-1 and apoA-2. The remaining forms of HDL are similar to nascent HDL. Subjects suffering from LCAT deficiency (all and part) have low HDL cholesterol.

[0013] Familial LCAT deficiency is a rare genetic disorder in which patients lack LCAT activity and are at risk for progressive chronic kidney disease and in some cases kidney failure. Ichthyosis is a partial LCAT deficiency in which LCAT is unable to esterify, or convert the acid to the alkyl group, cholesterol in HDL particles. However, LCAT remains active on cholesterol particles in VLDL and LDL.

[0014] 3.3. Fish eye disease

[0015] Achthyosis, also known as partial LCAT deficiency, is a disorder that causes the clear front surface of the eye (cornea) to gradually become cloudy. The cloudiness usually first appears during adolescence or early adulthood and consists of small gray cholesterol spots (opacities) distributed across the cornea.

[0016] Fish eye disease is characterized by abnormalities such as impaired vision, plaques of fatty material, and dense opacities. Fish eye disease is an autosomal recessive genetic disease caused by mutations in the LCAT gene located on chromosome 16q22.1.

[0017] The LCAT gene mutation that causes fish eye disease can damage α-LCAT activity, thereby reducing the ability of this enzyme to attach cholesterol to HDL. This impaired mechanism of reducing cholesterol in the body can cause cloudy spots containing cholesterol in the cornea. It is not clear why cholesterol deposition only affects the cornea of ​​this disorder. Mutations that affect α-LCAT activity and β-LCAT activity can cause a related disorder called complete LCAT deficiency, which involves corneal cloudy spots and features that affect other parts of the body.

[0018] Currently, there is no specific treatment to correct LCAT deficiency, so treatment focuses on relieving symptoms. In severe cases of fish eye disease, a corneal transplant may be recommended.

[0019] There is a need for new methods for treating subjects suffering from ocular diseases, such as ocular diseases associated with lipid accumulation. 4. Summary of the Invention

[0021] On the one hand, the present disclosure provides a method for treating eye diseases using a lipid-binding protein-based complex, such as CER-001, for example, an eye disease associated with lipid accumulation (e.g., in a subject with ocular lipid deposition). Other lipid-binding protein-based complexes that can be used in the methods of the present disclosure include apomer, cargomer, and HDL-based complexes or HDL-based complexes, such as CSL-111, CSL-112, ETC-216, or defatted HDL. In some eye diseases, lipids may accumulate in or near the eyes (e.g., lipids may accumulate in the meibomian glands or lacrimal glands of the subject). Exemplary eye diseases associated with lipid accumulation that can be treated by the methods of the present disclosure include dry eye disease, such as dry eye disease associated with meibomian gland dysfunction or lacrimal gland dysfunction, blepharitis, uveitis, corneal diseases such as lipid keratopathy, dry macular degeneration (dry AMD), Stargardt disease, and Leber idiopathic stellate neuroretinitis, and eye diseases associated with LCAT defects, such as fish eye disease. In some embodiments, the use of lipid-binding protein complexes can reduce the severity of eye diseases. In some embodiments, the use of lipid-binding protein complexes can slow the progression of eye diseases. Without being bound by theory, it is believed that lipid-binding protein complexes can reduce ocular lipid deposits, such as by dissolving lipids accumulated in ocular deposits, thereby causing them to be eliminated.

[0022] On the other hand, the present disclosure provides a method for treating an eye disease by delivering an ophthalmic drug to the eye of a subject suffering from an eye disease using a lipid-binding protein-based complex (e.g., CER-001) as a drug carrier. For example, the subject can be a subject suffering from an anterior or posterior eye disease, such as uveitis, macular edema (e.g., diabetic macular edema), macular degeneration, retinal detachment, eye tumors, fungal or viral infections, multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis, or glaucoma.

[0023] In another aspect, the present disclosure provides a composition comprising a lipid binding protein-based complex (eg, CER-001) and one or more ophthalmic drugs complexed therewith.

[0024] In the methods described herein, the lipid-binding protein-based complex (e.g., CER-001) can be administered systemically (e.g., by infusion). Alternatively, the lipid-binding protein-based complex (e.g., CER-001) can be administered locally (e.g., by intraocular or topical administration). Intraocular administration can be by, for example, intraocular injection, such as intravitreal injection, subconjunctival injection, juxtabular injection, peribulbar injection, or retrobulbar injection. For topical administration, the lipid-binding protein-based complex (e.g., CER-001) can be administered, for example, as eye drops. Other delivery methods can also be used, such as delivery via suprachoroidal injection, suprascleral injection, implants (e.g., discs, sheets, plugs, rods, or pellets), iontophoresis, or electroporation.

[0025] In one aspect, the present disclosure provides a dosing regimen of a lipid-binding protein-based complex (e.g., CER-001) for treating a subject with an eye disease associated with lipid accumulation. The dosing regimens described herein can also be applied to delivering ophthalmic drugs to the eye using a lipid-binding protein-based complex (e.g., CER-001) as a drug carrier.

[0026] In some embodiments, the dosing regimen of the present disclosure administers a lipid-binding protein-based complex (e.g., CER-001) to a subject according to an initial "induction" regimen, then administers a lipid-binding protein-based complex (e.g., CER-001) to a subject according to a "consolidation" regimen, and then administers a lipid-binding protein-based complex (e.g., CER-001) to a subject according to a "maintenance" regimen. In addition, the dosing regimen may administer a lipid-binding protein-based complex (e.g., CER-001) to a subject according to a "maintenance" regimen without including a previous "induction" regimen or "consolidation" regimen. Alternatively, the dosing regimen may administer a lipid-binding protein-based complex (e.g., CER-001) to a subject according to an "induction" regimen and a "maintenance" regimen without an intermediate "consolidation" regimen.

[0027] The induction regimen generally includes administering multiple doses of a lipid-binding protein-based complex (e.g., CER-001) to the subject, with 1 day or more between each dose. In some embodiments, the induction regimen includes three or more doses of a lipid-binding protein-based complex (e.g., CER-001). In some embodiments, the induction regimen includes three doses of a lipid-binding protein-based complex (e.g., CER-001) per week. In some embodiments, the induction regimen includes three doses of a lipid-binding protein-based complex (e.g., CER-001) per week for a period of more than one week, for example, two weeks or longer. In some embodiments, the induction regimen includes three doses of a lipid-binding protein-based complex (e.g., CER-001) per week for a period of three weeks.

[0028] The consolidation scheme generally includes administering multiple doses of lipid-binding protein-based complexes (e.g., CER-001) to the subject at a frequency lower than during the induction scheme. The consolidation scheme generally includes administering multiple doses of lipid-binding protein-based complexes (e.g., CER-001) to the subject, with 1 day or more between each dose, for example, 2 days or more between each dose. In some embodiments, the consolidation scheme includes two or more doses of lipid-binding protein-based complexes (e.g., CER-001). In some embodiments, the consolidation scheme includes two doses of lipid-binding protein-based complexes (e.g., CER-001) per week. In some embodiments, the consolidation scheme includes two doses of lipid-binding protein-based complexes (e.g., CER-001) per week for more than a week, for example, two weeks or longer. In some embodiments, the consolidation scheme includes two doses of lipid-binding protein-based complexes (e.g., CER-001) per week for three weeks.

[0029] The maintenance regimen generally includes administering one or more doses of a lipid-binding protein-based complex (e.g., CER-001) to the subject at a lower frequency than during the consolidation regimen, such as with an interval of 5 days or more between doses, e.g., a week's time. In certain embodiments, multiple doses of a lipid-binding protein-based complex (e.g., CER-001) are administered once a week during the maintenance regimen.

[0030] In certain aspects, the present disclosure provides a method of treating a subject with a lipid-binding protein-based complex (e.g., CER-001) using an induction regimen, the induction regimen comprising administering to the subject three doses of the lipid-binding protein-based complex (e.g., CER-001) in one week for three weeks with at least 1 day between each dose, followed by a consolidation regimen comprising administering to the subject two doses of the lipid-binding protein-based complex (e.g., CER-001) in one week for three weeks with at least 2 days between each dose, followed by a maintenance regimen comprising administering to the subject one dose of the lipid-binding protein-based complex (e.g., CER-001) per week.

[0031] In certain aspects, the present disclosure provides a method of treating a subject with a lipid-binding protein-based complex (e.g., CER-001) according to a dosing regimen described herein. In some embodiments, the lipid-binding protein-based complex (e.g., CER-001) is diluted with saline before intravenous administration (e.g., intravenous infusion using an infusion pump). In certain embodiments, the dosage of the lipid-binding protein-based complex (e.g., CER-001) for infusion is based on the subject's body weight, such as 10 mg / kg based on protein weight.

[0032] In certain aspects, the disclosure provides methods of treating a subject suffering from an ocular disease (e.g., associated with lipid accumulation) with a lipid binding protein-based complex (e.g., CER-001) according to a dosing regimen comprising:

[0033] -3 doses per week for 3 weeks (induction regimen), then

[0034] -2 doses per week for 3 weeks (consolidation regimen), then

[0035] - 1 dose per week until the end of treatment (maintenance regimen).

[0036] In certain aspects, an antihistamine (e.g., dexchlorpheniramine, hydroxyzine, diphenhydramine, cetirizine, fexofenadine, or loratadine) can be administered prior to administration of a lipid-binding protein-based complex (e.g., CER-001), e.g., when the lipid-binding protein-based complex is administered by IV infusion. Antihistamines can reduce the likelihood of allergic reactions.

[0037] The experimenter of the treatment of dosage regimen disclosed herein can be any experimenter suffering from the ophthalmopathy relevant to lipid accumulation, for example, the experimenter suffering from LCAT defect.LCAT defect can be complete LCAT defect or part LCAT defect.In some embodiments, the experimenter of the treatment of dosage regimen disclosed herein suffers from fish eye disease.Or, the experimenter of the treatment of dosage regimen disclosed herein can also be any experimenter of ophthalmic drug treatment that needs, wherein use the complex (for example, CER-001) based on lipid-binding protein as drug carrier by drug delivery to eyes. 5. Description of the drawings

[0038] Figures 1A-1D : Shows CER-001 as an ophthalmic drug azithromycin ( Figure 1A ), spironolactone ( Figure 1B ), dexamethasone palmitate ( Figure 1C ) and cyclosporine ( Figure 1D ) as a drug carrier.

[0039] Figures 2A-2C : shows the tolerability scores of rabbits administered CER-001, with or without complexed dexamethasone palmitate (Example 4). Figure 2A : 6-hour and 24-hour tolerance chart; Figure 2B : 6 hours of tolerance; Figure 2C : 24 hours tolerance.

[0040] Figure 3A-3B : shows cellular infiltration in the aqueous humor of rabbits administered CER-001, with or without complexed dexamethasone palmitate ( Figure 3A ) and protein ( Figure 3B )(Example 4).

[0041] Figure 4 : Shows the opacity of rabbit eyes administered CER-001 and soft corticosteroid (CER-S Cort) at 0 hours, 6 hours and 24 hours after administration. 6. Detailed description of the invention

[0043] In some aspects, the present disclosure provides methods for treating eye diseases (e.g., eye diseases associated with lipid accumulation) using lipid-binding protein-based complexes (e.g., CER-001). The methods of the present disclosure can reduce the severity of eye diseases in subjects. In some embodiments, the lipid-binding protein-based complex is an apomer, a cargomer, a HDL-based complex, or a HDL-mimic-based complex. In some embodiments, the lipid-binding protein-based complex can be used as a drug carrier to deliver one or more ophthalmic drugs to the eye, such as one or more hydrophobic and / or poorly water-soluble or water-insoluble ophthalmic drugs.

[0044] In some embodiments, the lipid binding protein-based complex (e.g., CER-001) (e.g., when used as a drug carrier or not) does not contain and is not administered with a cell penetrating peptide (CPP) (e.g., a CPP as described in WO 2019 / 018350), a chemical permeation enhancer (CPE) (e.g., a CPE as described in WO 2019 / 018350), or a cytopathic peptide (e.g., a cytopathic peptide as described in EP 3 238 746A1). The contents of WO 2019 / 018350 and EP 3238 746A1 are incorporated herein by reference in their entirety.

[0045] In other aspects, the present disclosure provides lipid-binding protein-based complexes, such as CER-001, used as carriers for one or more ophthalmic drugs. Thus, in some aspects, the present disclosure provides compositions comprising lipid-binding protein-based complexes (e.g., CER-001) and one or more ophthalmic drugs complexed therewith (e.g., as described in Section 6.1.9). Such compositions can be used in the methods of the present disclosure.

[0046] Exemplary features of lipid binding protein-based complexes that can be used in the methods and compositions of the present disclosure are described in Section 6.1. Exemplary subject populations that can be treated by the methods of the present disclosure and the compositions of the present disclosure are described in Section 6.2.

[0047] The lipid-binding protein-based complex can be administered peripherally or topically. In some embodiments, the lipid-binding protein-based complex is administered peripherally, such as by infusion. In other embodiments, the lipid-binding protein-based complex is administered topically (e.g., by intraocular or topical administration).

[0048] In some embodiments, the methods of the present disclosure include administering a lipid-binding protein-based complex (e.g., CER-001) to a subject in three stages. First, the lipid-binding protein-based complex (e.g., CER-001) is administered with an initial, strong "induction" regimen. The induction regimen is followed by a "consolidation" regimen of lower intensity. The consolidation regimen is followed by a "maintenance" regimen. In other methods of the present disclosure, the lipid-binding protein-based complex (e.g., CER-001) is administered in two stages (e.g., an induction regimen is followed by a maintenance regimen) or a single stage (e.g., a maintenance regimen). The induction regimen that can be used in the methods of the present disclosure is described in Section 6.3, the consolidation regimen that can be used in the methods of the present disclosure is described in Section 6.4, and the maintenance regimen that can be used in the methods of the present disclosure is described in Section 6.5. The dosing regimen of the present disclosure includes administering a lipid-binding protein-based complex (e.g., CER-001) as a monotherapy or as part of a combination therapy with one or more drugs. Combination therapy is described in Section 6.6.

[0049] 6.1. Lipid-binding protein-based complexes

[0050] 6.1.1. Composites based on HDL and HDL mimetics

[0051] On the one hand, the lipid-binding protein-based complex comprises a complex based on HDL or HDL mimics. For example, the complex may include a lipoprotein complex as described in U.S. Patent No. 8,206,750, PCT Publication WO 2012 / 109162, PCT Publication WO 2015 / 173633 A2 (e.g., CER-001), PCT Publication WO 2004 / 073684 or US2004 / 0229794 A1, the contents of each of which are incorporated herein by reference in their entirety. The terms "lipoprotein" and "apolipoprotein" are used interchangeably herein, unless the context otherwise requires, otherwise the term "lipoprotein" covers lipoprotein mimics. The terms "lipid-binding protein" and "lipid-binding polypeptide" are also used interchangeably herein, unless the context otherwise requires, otherwise these terms do not represent amino acid sequences of specific lengths.

[0052] The lipoprotein complex can comprise a protein portion (e.g., an apolipoprotein portion) and a lipid portion (e.g., a phospholipid portion). The protein portion includes one or more lipid binding protein molecules, such as apolipoprotein, a peptide, or an apolipoprotein peptide analog or mimetic, such as one or more lipid binding protein molecules described in Section 6.1.4. In some embodiments, the lipid binding protein molecules include apolipoprotein molecules (e.g., ApoA-I molecules), but do not include apolipoprotein mimetic molecules.

[0053] The lipid fraction generally includes one or more phospholipids that can be neutral, negatively charged, positively charged, or a combination thereof. Exemplary phospholipids and other amphiphilic molecules that can be included in the lipid fraction are described in Section 6.1.6.

[0054] In certain embodiments, the lipid portion contains at least one neutral phospholipid (e.g., sphingomyelin (SM)) and optionally one or more negatively charged phospholipids. In lipoprotein complexes that include both neutral and negatively charged phospholipids, the neutral and negatively charged phospholipids can have fatty acid chains with the same or different carbon numbers and the same or different degrees of saturation. In some cases, the neutral and negatively charged phospholipids have the same acyl tail, such as C16:0, or palmitoyl, acyl chains. In specific embodiments, especially those using egg SM as a neutral lipid, the weight ratio of the apolipoprotein portion:lipid portion ranges from about 1:2.7 to about 1:3 (e.g., 1:2.7).

[0055] Any phospholipid that carries at least a partial negative charge at physiological pH can be used as a negatively charged phospholipid. Non-limiting examples include negatively charged forms of phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, and phosphatidic acid, for example, salts. In a specific embodiment, the negatively charged phospholipid is 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)], or DPPG, phosphatidylglycerol. Preferred salts include potassium salts and sodium salts.

[0056] In some embodiments, the lipoprotein complex used in the compositions and methods of the present disclosure is a lipoprotein complex as described in U.S. Pat. No. 8,206,750 or WO 2012 / 109162 (and its U.S. equivalent US2012 / 0232005), the contents of each of which are incorporated herein by reference in their entirety. In specific embodiments, the protein component of the lipoprotein complex is as described in Section 6.1 and preferably Section 6.1.1 of WO 2012 / 109162 (and US 2012 / 0232005), and the lipid component is as described in Section 6.2 of WO 2012 / 109162, which can optionally be compounded together in the amounts described in Section 6.3 of WO 2012 / 109162 (and US 2012 / 0232005). The contents of each of these sections are incorporated herein by reference. In certain aspects, the lipoprotein complexes of the present disclosure are at least 85%, at least 90%, at least 95%, at least 97% or at least 99% homogeneous in a population of complexes, as described in Section 6.4 of WO 2012 / 109162 (and US 2012 / 0232005), the contents of which are incorporated herein by reference.

[0057] In specific embodiments, lipoprotein complexes useful in the compositions and methods of the present disclosure contain 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50-80 phosphatidylcholine molecules, and 20-50 SM molecules.

[0058] In another specific embodiment, the lipoprotein complex that can be used in the compositions and methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50 phosphatidylcholine molecules, and 50 SM molecules.

[0059] In yet another specific embodiment, the lipoprotein complex that can be used in the compositions and methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 80 phosphatidylcholine molecules, and 20 SM molecules.

[0060] In yet another specific embodiment, the lipoprotein complex that can be used in the compositions and methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 70 phosphatidylcholine molecules, and 30 SM molecules.

[0061] In yet another specific embodiment, the lipoprotein complex that can be used in the compositions and methods of the present disclosure comprises 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 60 phosphatidylcholine molecules, and 40 SM molecules.

[0062] In specific embodiments, the lipoprotein complexes useful in the compositions and methods of the present disclosure consist essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50-80 phosphatidylcholine molecules, and 20-50 SM molecules.

[0063] In another specific embodiment, the lipoprotein complex useful in the compositions and methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 50 phosphatidylcholine molecules, and 50 SM molecules.

[0064] In yet another specific embodiment, the lipoprotein complex useful in the compositions and methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 80 phosphatidylcholine molecules, and 20 SM molecules.

[0065] In yet another specific embodiment, the lipoprotein complex useful in the compositions and methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 70 phosphatidylcholine molecules, and 30 SM molecules.

[0066] In yet another specific embodiment, the lipoprotein complex useful in the compositions and methods of the present disclosure consists essentially of 2-4 ApoA-I equivalents, 2 charged phospholipid molecules, 60 phosphatidylcholine molecules, and 40 SM molecules.

[0067] In specific embodiments, the lipoprotein complexes that can be used in the compositions and methods of the present disclosure comprise a lipid component comprising about 90 to 99.8 wt % SM and about 0.2 to 10 wt % negatively charged phospholipids, for example, about 0.2-1 wt %, 0.2-2 wt %, 0.2-3 wt %, 0.2-4 wt %, 0.2-5 wt %, 0.2-6 wt %, 0.2-7 wt %, 0.2-8 wt %, 0.2-9 wt % or 0.2-10 wt % total negatively charged phospholipids. In another specific embodiment, the lipoprotein complexes that can be used in the methods of the present disclosure contain about 90 to 99.8 wt % phosphatidylcholine and about 0.2 to 10 wt % negatively charged phospholipids, for example, about 0.2-1 wt %, 0.2-2 wt %, 0.2-3 wt %, 0.2-4 wt %, 0.2-5 wt %, 0.2-6 wt %, 0.2-7 wt %, 0.2-8 wt %, 0.2-9 wt % or 0.2-10 wt % of the total negatively charged phospholipids.

[0068] In specific embodiments, the lipoprotein complexes that can be used in the compositions and methods of the present disclosure comprise a lipid component that consists essentially of about 90 to 99.8 wt% SM and about 0.2 to 10 wt% negatively charged phospholipids, for example, about 0.2-1 wt%, 0.2-2 wt%, 0.2-3 wt%, 0.2-4 wt%, 0.2-5 wt%, 0.2-6 wt%, 0.2-7 wt%, 0.2-8 wt%, 0.2-9 wt% or 0.2-10 wt% total negatively charged phospholipids. In another specific embodiment, the lipoprotein complexes that can be used in the methods of the present disclosure consist essentially of about 90 to 99.8 wt % phosphatidylcholine and about 0.2 to 10 wt % negatively charged phospholipids, for example, about 0.2-1 wt %, 0.2-2 wt %, 0.2-3 wt %, 0.2-4 wt %, 0.2-5 wt %, 0.2-6 wt %, 0.2-7 wt %, 0.2-8 wt %, 0.2-9 wt % or 0.2-10 wt % of the total negatively charged phospholipids.

[0069] In yet another specific embodiment, the lipoprotein complex that can be used in the compositions and methods of the present disclosure comprises a lipid portion comprising about 9.8 to 90 wt % SM, about 9.8 to 90 wt % phosphatidylcholine, and about 0.2-10 wt % negatively charged phospholipids, for example, from about 0.2-1 wt %, 0.2-2 wt %, 0.2-3 wt %, 0.2-4 wt %, 0.2-5 wt %, 0.2-6 wt %, 0.2-7 wt %, 0.2-8 wt %, 0.2-9 wt % to 0.2-10 wt % total negatively charged phospholipids.

[0070] In yet another specific embodiment, the lipoprotein complexes that can be used in the compositions and methods of the present disclosure comprise a lipid portion consisting essentially of about 9.8 to 90 wt % SM, about 9.8 to 90 wt % phosphatidylcholine, and about 0.2-10 wt % negatively charged phosphatidylcholine, for example, from about 0.2-1 wt %, 0.2-2 wt %, 0.2-3 wt %, 0.2-4 wt %, 0.2-5 wt %, 0.2-6 wt %, 0.2-7 wt %, 0.2-8 wt %, 0.2-9 wt % to 0.2-10 wt % total negatively charged phospholipids.

[0071] In another specific embodiment, the lipoprotein complex that can be used in the compositions and methods of the present disclosure comprises an ApoA-I apolipoprotein and a lipid portion, wherein the lipid portion comprises sphingomyelin and about 3 wt % of a negatively charged phospholipid, wherein the molar ratio of the lipid portion to the ApoA-I apolipoprotein is about 2:1 to 200:1, wherein the complex is a small or large disc-shaped particle containing 2-4 ApoA-I equivalents.

[0072] In another specific embodiment, the lipoprotein complex that can be used in the compositions and methods of the present disclosure comprises an ApoA-I apolipoprotein and a lipid portion, wherein the lipid portion consists essentially of sphingomyelin and about 3 wt % of negatively charged phospholipids, wherein the molar ratio of the lipid portion to the ApoA-I apolipoprotein is about 2:1 to 200:1, and wherein the complex is a small or large disc-shaped particle containing 2-4 ApoA-I equivalents.

[0073] HDL-based or HDL-mimic-based complexes can include a single type of lipid-binding protein, or a mixture of two or more different lipid-binding proteins, which can be derived from the same or different species. Although not required, the complex will preferably contain a lipid-binding protein derived from the treated animal species or corresponding to the treated animal species in amino acid sequence to avoid inducing an immune response to the therapy. Therefore, for the treatment of human patients, lipid-binding proteins of human origin are preferably used. The use of peptide mimics of apolipoproteins can also reduce or avoid immune responses.

[0074] In some embodiments, the lipid component includes two types of phospholipids: sphingomyelin (SM) and negatively charged phospholipids. Exemplary SM and negatively charged lipids are described in Section 6.1.6.1.

[0075] The lipid component comprising SM may optionally include a small amount of additional lipids. Virtually any type of lipid may be used, including but not limited to lysophospholipids, galactocerebrosides, gangliosides, cerebrosides, glycerides, triglycerides and cholesterol and their derivatives.

[0076] When included, such optional lipids will generally account for less than about 15 wt% of the lipid portion, although more optional lipids may be included in some cases. In some embodiments, the optional lipids account for less than about 10 wt%, less than about 5 wt%, or less than about 2 wt%. In some embodiments, the lipid portion does not include optional lipids.

[0077] In a specific embodiment, the phospholipid fraction contains egg SM or palmitoyl SM or phytosphingomyelin and DPPG in a weight ratio (SM: negatively charged phospholipid) ranging from 90:10 to 99:1, more preferably ranging from 95:5 to 98:2. In one embodiment, the weight ratio is 97:3.

[0078] The molar ratio of the lipid component to the protein component of the complex of the present disclosure can vary and will depend, among other factors, on the properties of the apolipoprotein comprising the protein component, the properties and quantity of the lipid comprising the lipid component, and the desired size of the complex. Because the biological activity of apolipoproteins, such as ApoA-I, is believed to be mediated by the amphipathic helices comprising the apolipoprotein, it is convenient to express the apolipoprotein portion of the lipid: apolipoprotein molar ratio using ApoA-I protein equivalents. It is generally believed that ApoA-I contains 6-10 amphipathic helices, depending on the method used to calculate the helices. Other apolipoproteins can be expressed in ApoA-I equivalents based on the number of amphipathic helices they contain. For example, ApoA-I, which is generally present in the form of a disulfide-bridged dimer, is preferably present in the form of a disulfide-bridged dimer. M It can be expressed as 2 ApoA-I equivalents, because each ApoA-I M The molecule contains twice as many amphipathic helices as an ApoA-I molecule. In contrast, a peptide apolipoprotein containing a single amphipathic helix can be expressed as 1 / 10-1 / 6 ApoA-I equivalents, because each molecule contains 1 / 10-1 / 6 of the amphipathic helices of an ApoA-I molecule. Typically, the molar ratio of lipid: ApoA-I equivalents (defined herein as "Ri") of the lipoprotein complex ranges from about 105:1 to 110:1. In some embodiments, Ri is about 108:1. A MW of about 650-800 can be used to obtain a weight ratio for phospholipids.

[0079] In some embodiments, the lipid:ApoA-I equivalent molar ratio ("RSM") ranges from about 80: 1 to about 110: 1, such as about 80: 1 to about 100: 1. In a specific example, the RSM of the complex can be about 82:1.

[0080] In some embodiments, the lipoprotein complex used in the methods of the present disclosure is a negatively charged complex comprising a protein portion, preferably mature full-length ApoA-I, and a lipid portion comprising a neutral phospholipid, sphingomyelin (SM), and a negatively charged phospholipid.

[0081] In a specific embodiment, the lipid component contains SM (e.g., egg SM, palmitoyl SM, plant SM, or a combination thereof) and a negatively charged phospholipid (e.g., DPPG) in a weight ratio (SM: negatively charged phospholipid) ranging from 90:10 to 99:1, more preferably ranging from 95:5 to 98:2, for example 97:3.

[0082] In specific embodiments, the ratio of the protein component to the lipid component can range from about 1:2.7 to about 1:3, preferably 1:2.7. This corresponds to a molar ratio of ApoA-I protein to lipid ranging from about 1:90 to 1:140. In some embodiments, the molar ratio of protein to lipid in the complex is about 1:90 to about 1:120, about 1:100 to about 1:140, or about 1:95 to about 1:125.

[0083] In particular embodiments, the complex comprises CER-001, CSL-111, CSL-112, CER-522, or ETC-216. In preferred embodiments, the complex is CER-001.

[0084] CER-001 as used in the literature and in the following examples refers to the complex described in Example 4 of WO 2012 / 109162. WO 2012 / 109162 refers to CER-001 as a complex having a lipoprotein weight:total phospholipid weight ratio of 1:2.7 and a SM:DPPG weight:weight ratio of 97:3. Example 4 of WO 2012 / 109162 also describes its preparation method.

[0085] When used in the context of the CER-001 dosing regimen or composition of the present disclosure, CER-001 refers to a lipoprotein complex whose individual components may differ by up to 20% from the CER-001 described in Example 4 of WO 2012 / 109162. In certain embodiments, the components of the lipoprotein complex differ by up to 10% from the CER-001 as described in Example 4 of WO 2012 / 109162. Preferably, the components of the lipoprotein complex are those described in Example 4 of WO 2012 / 109162 (plus / minus acceptable variations in manufacturing tolerances). The SM in CER-001 may be natural or synthetic. In some embodiments, the SM is a natural SM, such as a natural SM described in WO 2012 / 109162, such as an egg SM. In some embodiments, the SM is a synthetic SM, such as a synthetic SM described in WO 2012 / 109162, such as a synthetic palmitoyl sphingomyelin, such as described in WO 2012 / 109162. Methods for synthesizing palmitoyl sphingomyelin are known in the art, for example as described in WO 2014 / 140787. The lipoprotein in CER-001, apolipoprotein AI (ApoA-I), preferably has an amino acid sequence corresponding to amino acids 25 to 267 of SEQ ID NO: 1 of WO 2012 / 109162 (SEQ ID NO: 1 of WO 2012 / 109162 is disclosed herein as SEQ ID NO: 2). ApoA-I can be purified from an animal source (particularly a human source) or recombinantly produced. In a preferred embodiment, the ApoA-I in CER-001 is recombinant ApoA-I. The CER-001 used in the dosing regimen of the present disclosure is preferably highly homogeneous, for example at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% homogeneous, as reflected by a single peak in gel permeation chromatography. See, for example, Section 6.4 of WO 2012 / 109162.

[0086] CSL-111 is a reconstituted human ApoA-I purified from plasma complexed with soybean phosphatidylcholine (SBPC) (Tardif et al., 2007, JAMA 297: 1675-1682).

[0087] CSL-112 is a preparation of ApoA-I purified from plasma and reconstituted to form HDL suitable for intravenous infusion (Diditchenko et al., 2013, DOI 10.1161 / ATVBAHA.113.301981).

[0088] ETC-216 (also known as MDCO-216) is a recombinant ApoA-I MilanoSee Nicholls et al., 2011, Expert Opin Biol Ther. 11(3):387-94. doi:10.1517 / 14712598.2011.557061.

[0089] In another embodiment, the complex that can be used in the methods of the present disclosure is CER-522. CER-522 is a lipoprotein complex that contains a combination of three phospholipids and a 22 amino acid peptide CT80522:

[0090]

[0091] CT80522

[0092] The phospholipid component of CER-522 is composed of egg sphingomyelin, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (dipalmitoylphosphatidylcholine, DPPC) and 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (dipalmitoylphosphatidylglycerol, DPPG) in a weight ratio of 48.5:48.5:3. The ratio of peptide to total phospholipids in the CER-522 complex is 1:2.5 (w / w).

[0093] In some embodiments, the lipoprotein complex is delipidated HDL. Most HDL in plasma is rich in cholesterol. Lipids in HDL can be reduced, such as partially and / or selectively reduced, for example, to reduce its cholesterol content. In some embodiments, delipidated HDL can be similar to small alpha, pre-beta-1 and other pre-beta forms of HDL. The process of selectively reducing HDL is described in Sacks et al., 2009 J Lipid Res. 50 (5): 894–907.

[0094] In certain embodiments, the lipoprotein complex comprises a bioactive agent delivery particle as described in US 2004 / 0229794.

[0095] The bioactive agent delivery particle can comprise a lipid-binding polypeptide (e.g., an apolipoprotein as described previously in this section or in Section 6.1.4), a lipid bilayer (e.g., comprising one or more phospholipids as described previously in this section or in Section 6.1.6.1), and a bioactive agent (e.g., an anticancer agent), wherein the interior of the lipid bilayer comprises a hydrophobic region, and wherein the bioactive agent is associated with the hydrophobic region of the lipid bilayer. In some embodiments, the bioactive agent delivery particle is as described in US2004 / 0229794.

[0096] In some embodiments, the bioactive agent delivery particle does not comprise a hydrophilic core.

[0097] In some embodiments, the bioactive agent delivery particles are disc-shaped (eg, having a diameter of about 7 to about 29 nm).

[0098] The bioactive agent delivery particles include bilayer-forming lipids, such as phospholipids (e.g., as described previously in this section or Section 6.1.6.1). In some embodiments, the bioactive agent delivery particles include bilayer-forming and non-bilayer-forming lipids. In some embodiments, the lipid bilayer of the bioactive agent delivery particles includes phospholipids. In one embodiment, the phospholipids incorporated into the delivery particles include dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylglycerol (DMPG). In one embodiment, the lipid bilayer includes DMPC and DMPG in a molar ratio of 7:3.

[0099] In some embodiments, the lipid-binding polypeptide is an apolipoprotein (e.g., as described previously in this section or in Section 6.1.4). The primary interaction between the lipid-binding polypeptide (e.g., an apolipoprotein molecule) and the lipid bilayer is typically a hydrophobic interaction between residues on the hydrophobic face of the amphipathic structure (e.g., an α-helix of the lipid-binding polypeptide) and the fatty acyl chains of the lipids on the outer surface of the particle periphery. The bioactive agent delivery particle may include exchangeable and / or non-exchangeable apolipoproteins. In one embodiment, the lipid-binding polypeptide is ApoA-I.

[0100] In some embodiments, the bioactive agent delivery particle includes a lipid-binding polypeptide molecule, e.g., an apolipoprotein molecule, which has been modified to increase the stability of the particle. In one embodiment, the modification includes the introduction of cysteine ​​residues to form intramolecular and / or intermolecular disulfide bonds.

[0101] In another embodiment, the bioactive agent delivery particle comprises a chimeric lipid-binding polypeptide molecule, such as a chimeric apolipoprotein molecule, wherein one or more associated functional moieties, such as one or more targeting moieties and / or one or more moieties having a desired biological activity, such as antimicrobial activity, can enhance the activity of or act synergistically with the activity of the bioactive agent incorporated into the delivery particle.

[0102] 6.1.2. Apomer-based complexes

[0103] In one aspect, lipid-binding protein-based complexes useful in the methods and compositions of the present disclosure include apomers. Features of apomers that may be included in apomer-based complexes are described in WO / 2019 / 030575, the contents of which are incorporated herein by reference in their entirety.

[0104] Apomers typically include apolipoproteins in monomeric or polymeric form complexed with amphiphilic molecules. Typically, an apomer includes one or more apolipoprotein molecules, each apolipoprotein molecule complexed with one or more amphiphilic molecules. In certain aspects, the amphiphilic molecules contribute together to a net charge of at least +1 or -1 for each apolipoprotein molecule in the apomer. Exemplary apolipoproteins that can be used for apomers are described in Section 6.1.5.1. Exemplary amphiphilic molecules are described in Section 6.1.6.

[0105] 6.1.3. Cargomer-based complexes

[0106] On the one hand, the lipid-binding protein-based complex that can be used for the methods and compositions of the present disclosure includes a Cargomer, which is a lipid-binding protein-based complex with one or more cargo portions. The characteristics of the Cargomer that can be included in the Cargomer-based complex are described in WO / 2019 / 030574, the contents of which are incorporated herein by reference in their entirety.

[0107] Cargomers typically comprise apolipoproteins (e.g., 2, 4, or 8 apolipoprotein molecules) in monomeric or polymeric form and one or more cargo moieties. The cargo moiety may be amphiphilic or non-amphiphilic. The amphiphilic cargo moiety may dissolve apolipoproteins and prevent them from aggregating. When the cargo moiety is not amphiphilic or insufficient to dissolve the apolipoprotein molecules, the cargomer may further comprise one or more additional amphiphilic molecules to dissolve the apolipoproteins. Therefore, amphiphilic molecules mentioned in the context of cargomers include amphiphilic molecules as cargo moieties, amphiphilic molecules that are not cargo moieties, or some combination thereof. Preferably, the cargomer is not disc-shaped, as determined, for example, using NMR spectroscopy.

[0108] The cargo portion may include biologically active molecules (e.g., drugs, biologics, and / or immunogens) or other agents, such as agents for diagnosis. As used herein, the terms "molecule" and "agent" also include complexes and conjugates (e.g., antibody-drug conjugates). The terms "biologically active," "diagnostically useful," and the like are not limited to substances with direct pharmacological or biological activity, and may include substances that become active after administration, such as due to metabolism of a prodrug or cleavage of a linker. Therefore, the terms "biologically active" and "diagnostically useful" also include substances that become biologically active or diagnostically useful after administration by producing or metabolites or other cleavage products that exert pharmacological or biological effects and / or detectable in diagnostic tests.

[0109] The amphiphilic molecules in Cargomer can dissolve apolipoprotein and / or reduce or minimize apolipoprotein aggregation, and can also have other functions in Cargomer. For example, amphiphilic molecules can have therapeutic utility, and therefore can be the cargo portion intended to be delivered by Cargomer when administered to a subject. In addition, as discussed below in Section 6.1.6, amphiphilic molecules can be used to anchor non-amphiphilic cargo portions to apolipoprotein in Cargomer. Therefore, in some embodiments, the cargo portion and amphiphilic molecules in Cargomer are the same. In other embodiments, the anchoring portion and amphiphilic molecules in Cargomer are the same. In another other embodiment, the cargo portion, anchoring portion and amphiphilic molecules in Cargomer are the same (for example, wherein the amphiphilic molecules have therapeutic activity and also anchor another biologically active molecule to the apolipoprotein molecule).

[0110] Anchor and / or linker moieties are particularly useful for cargomers having a cargo moiety that is not an amphiphilic molecule.

[0111] In some embodiments, at least one cargo portion, most cargo portions, or all cargo portions in a cargomer of the present disclosure are coupled to a cargomer via an anchor. In some embodiments, at least one cargo portion in a cargomer is coupled to a cargomer via an anchor. In some embodiments, most cargo portions in a cargomer are coupled to a cargomer via an anchor. In some embodiments, all cargo portions in a cargomer are coupled to a cargomer via an anchor. Each anchor in a cargomer can be the same, or different types of anchors can be included in a single cargomer (e.g., one type of cargo portion can be coupled to a cargomer via one type of anchor and a second type of cargo portion can be coupled to a cargomer via a second type of anchor).

[0112] In certain aspects, the amphiphilic molecule, cargo, and anchor and / or linker (if present) together contribute a net charge of at least +1 or -1 (e.g., +1, +2, +3, -1, -2, or -3) to each apolipoprotein molecule in the Cargomer. In some embodiments, the net charge is a negative charge. In other embodiments, the net charge is a positive charge. Unless the context requires otherwise, the charge is measured at physiological pH.

[0113] The molar ratio of apolipoprotein molecules to amphiphilic molecules in the cargomer can be, but does not necessarily have to be, an integer or reflect a one-to-one relationship between apolipoprotein and amphiphilic molecules. As an example and not limitation, the cargomer can have a molar ratio of apolipoprotein to amphiphilic molecules of 2:5, 8:7, 3:2 or 4:7.

[0114] In some embodiments, the cargomer comprises an apolipoprotein molecule complexed with an amphiphile molecule, and the molar ratio of apolipoprotein:amphiphile molecule ranges from 8:1 to 1:15 (e.g., from 8:1 to 1:15, from 7:1 to 1:15, from 6:1 to 1:15, from 5:1 to 1:15, from 4:1 to 1:15, from 3:1 to 1:15, from 2:1 to 1:15, from 1:1 to 1:15, from 8:1 to 1:14, from 7:1 to 1:14, from 6:1 to 1:14, from 5:1 to 1:14, from 4:1 to 1:14, from 3:1 to 1:14, from 2:1 to 1:14, from 1:1 to 1:14, from 8:1 to 1:13, from 7:1 to 1:13, from 6:1 to 1:1 3, from 5:1 to 1:13, from 4:1 to 1:13, from 3:1 to 1:13, from 2:1 to 1:13, from 1:1 to 1:13, from 8:1 to 1:12, from 7:1 to 1:12, from 6:1 to 1:12, from 5:1 to 1:12, from 4:1 to 1:12, from 3:1 to 1:12, from 2:1 to 1:12, from 1: 1 to 1:12, from 8:1 to 1:11, from 7:1 to 1:11, from 6:1 to 1:11, from 5:1 to 1:11, from 4:1 to 1:11, from 3:1 to 1:11, from 2:1 to 1:11, from 1:1 to 1:11, from 8:1 to 1:10, from 7:1 to 1:10, from 6:1 to 1:10, from 5:1 to 1:1 0, from 4:1 to 1:10, from 3:1 to 1:10, from 2:1 to 1:10, from 1:1 to 1:10, from 8:1 to 1:9, from 7:1 to 1:9, from 6:1 to 1:9, from 5:1 to 1:9, from 4:1 to 1:9, from 3:1 to 1:9, from 2:1 to 1:9, from 1:1 to 1:9, from 8:1 to 1:8, from 7 :1 to 1:8, from 6:1 to 1:8, from 5:1 to 1:8, from 4:1 to 1:8, from 3:1 to 1:8, from 2:1 to 1:8, from 1:1 to 1:8, from 8:1 to 1:7, from 7:1 to 1:7, from 6:1 to 1:7, from 5:1 to 1:7, from 4:1 to 1:7, from 3:1 to 1:7, from 2:1 to 1:7, from 1:1 to 1:7, from 8:1 to 1:6, from 7:1 to 1:6, from 6:1 to 1:6, from 5:1 to 1:6, from 4:1 to 1:6, from 3:1 to 1:6, from 2:1 to 1:6, from 1:1 to 1:6, from 8:1 to 1:5, from 7:1 to 1:5, from 6:1 to 1:5, from 5:1 to 1:5, from 4:1 to 1:5, From 3:1 to 1:5, from 2:1 to 1:5, from 1:1 to 1:5, from 8:1 to 1:4, from 7:1 to 1:4, from 6:1 to 1:4, from 5:1 to 1:4, from 4:1 to 1:4, from 3:1 to 1:4, from 2:1 to 1:4, from 1:1 to 1:4, from 8:1 to 1:3, from 7:1 to 1:3, from 6:1 to 1:3,from 5:1 to 1:3, from 4:1 to 1:3, from 3:1 to 1:3, from 2:1 to 1:3, from 1:1 to 1:3, from 8:1 to 1:2, from 7:1 to 1:2, from 6:1 to 1:2, from 5:1 to 1:2, from 4:1 to 1:2, from 3:1 to 1:2, from 2:1 to 1:2, from 1:1 to 1:2, from 8:1 to 1:1, from 7:1 to 1:1, from 6:1 to 1:1, from 5:1 to 1:1, from 4:1 to 1:1, from 3:1 to 1:1 or from 2:1 to 1:1).

[0115] In some embodiments, the molar ratio of apolipoprotein to amphiphile in Cargomer ranges from 6:1 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 5:1 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 4:1 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 3:1 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 2:1 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 5:1 to 1:5. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 4:1 to 1:5. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 3:1 to 1:5. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 2:1 to 1:5. In some embodiments, the molar ratio of apolipoprotein to amphiphile ranges from 5:1 to 1:4. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 4:1 to 1:4. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 3:1 to 1:4. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 2:1 to 1:4. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 5:1 to 1:3. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 4:1 to 1:3. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 3:1 to 1:3. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 2:1 to 1:3. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 5:1 to 1:2. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 4:1 to 1:2. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 3:1 to 1:2. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 2:1 to 1:2. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 5:1 to 1:1. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 4:1 to 1:1. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 3:1 to 1:1. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 2:1 to 1:1. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 1:1 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 1:1 to 1:5. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 1:1 to 1:4. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 1:1 to 1:3. In some embodiments, the molar ratio of apolipoprotein to amphiphile is in the range of 1:1 to 1:2.In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:2 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:2 to 1:5. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:2 to 1:4. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:2 to 1:3. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:3 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:3 to 1:5. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:3 to 1:4. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:4 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:4 to 1:5. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecules ranges from 1:5 to 1:6. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecule ranges from 1.5:1 to 1:2. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecule ranges from 5:4 to 4:5. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecule ranges from 5:3 to 3:5. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecule ranges from 5:2 to 2:5. In some embodiments, the molar ratio of apolipoprotein to amphiphilic molecule ranges from 3:2 to 2:3.

[0116] In some embodiments, the ratio of apolipoprotein molecules to amphiphilic molecules is about 1: 1. In other embodiments, the ratio of apolipoprotein molecules to amphiphilic molecules is about 1: 2. In other embodiments, the ratio of apolipoprotein molecules to amphiphilic molecules is about 1: 3. In other embodiments, the ratio of apolipoprotein molecules to amphiphilic molecules is about 1: 4. In other embodiments, the ratio of apolipoprotein molecules to amphiphilic molecules is about 1: 5. In other embodiments, the ratio of apolipoprotein molecules to amphiphilic molecules is about 1: 6.

[0117] In some embodiments, the cargomer comprises 1 apolipoprotein molecule.

[0118] In other embodiments, Cargomer comprises 2 apolipoprotein molecules. Cargomer comprising 2 apolipoprotein molecules preferably has a Stokes radius of 3 nm or less. In some embodiments, Cargomer may comprise 2 apolipoprotein molecules and 1, 2 or 3 negatively charged amphiphilic molecules (e.g., negatively charged phospholipid molecules) per apolipoprotein molecule.

[0119] In other embodiments, Cargomer comprises 4 apolipoprotein molecules. Cargomer comprising 4 apolipoprotein molecules preferably has a Stokes radius of 4 nm or less. In some embodiments, Cargomer may comprise 4 apolipoprotein molecules and 1, 2 or 3 negatively charged amphiphilic molecules (e.g., negatively charged phospholipid molecules) per apolipoprotein molecule.

[0120] In other embodiments, Cargomer comprises 8 apolipoprotein molecules. Cargomer comprising 8 apolipoprotein molecules preferably has a Stokes radius of 5nm or less. In some embodiments, Cargomer may comprise 8 apolipoprotein molecules and 1, 2 or 3 negatively charged amphiphilic molecules (e.g., negatively charged phospholipid molecules) per apolipoprotein molecule. In certain embodiments, Cargomer of the present disclosure does not contain cholesterol and / or cholesterol derivatives (e.g., cholesterol esters).

[0121] In some embodiments, the cargomer comprises a weight ratio of apolipoprotein to phospholipid in a range of about 1:2 to about 1:3.

[0122] In some embodiments, the cargomer comprises apolipoprotein to phospholipid in a weight ratio of 1:2.7.

[0123] The cargomer may be soluble in one or more of biological fluids, such as lymph, cerebrospinal fluid, vitreous humor, aqueous humor, and blood or blood components (eg, serum or plasma).

[0124] Cargomers can include targeting functionality, such as targeting the cargomer to a specific cell or tissue type. In some embodiments, the cargomer includes a targeting moiety attached to an apolipoprotein molecule or an amphiphilic molecule. In some embodiments, one or more cargo moieties incorporated into the cargomer have targeting capabilities.

[0125] 6.1.4. ApoA-I preparations

[0126] In one aspect, the present disclosure relates to an ApoA-I preparation comprising ApoA-I and one or more lipids, wherein the ApoA-I and the lipid are present in the form of a lipoprotein complex.

[0127] The ApoA-I can be any such apolipoprotein described in Section 6.1.5.1, including, inter alia, ApoA-I having the amino acid sequence of amino acids 25-267 of SEQ ID NO: 2 and / or recombinantly expressed ApoA-I.

[0128] The lipid may be any one or more of those described in Section 6.1.6.1. The lipid may include a neutral lipid and / or a negatively charged lipid. The neutral lipid may comprise or consist of a sphingomyelin, such as a natural sphingomyelin (e.g., egg sphingomyelin) and / or a synthetic sphingomyelin (e.g., palmitoyl sphingomyelin). The negatively charged lipid may comprise 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof, or consist of DPPG or a salt thereof.

[0129] Lipid can comprise any weight or molar ratio of neutral phospholipid and negatively charged phospholipid as described herein.Lipid can be by 95% to 99% weight neutral phospholipid and 1% to 5% weight negatively charged phospholipid, as 96% to 98% weight neutral phospholipid and 2% to 4% weight negatively charged phospholipid, or 97% weight neutral phospholipid and 3% weight negatively charged phospholipid composition.

[0130] The formulation can include ApoA-I and lipids in any weight or molar ratio described herein. For example, the molar ratio of negatively charged lipids to neutral lipids to ApoA-I components in the formulation is 2-6:90-120:1. Exemplary formulations can include ApoA-I to lipid weight ratios ranging from 1:2 to 1:3, such as about 1:2.7.

[0131] Optionally, the formulation may further comprise one or more ophthalmic drugs as described in Section 6.1.9.

[0132] The formulation can be used in a method of treating an eye disease, such as the methods described in Sections 6.2-6.6. The formulation can also be used to treat an eye disease. The formulation can also be used to prepare a medicament for treating an eye disease.

[0133] Examples of ApoA-I preparations and uses thereof include those listed as Group 1 numbered embodiments 357-458.

[0134] 6.1.5. Lipid binding protein molecules

[0135] Lipid binding protein molecules that can be used in the complexes described herein include apolipoproteins (as described in Section 6.1.5.1) and apolipoprotein mimetic peptides (as described in Section 6.1.5.2). In some embodiments, the complex comprises a mixture of lipid binding protein molecules. In some embodiments, the complex comprises a mixture of one or more lipid binding protein molecules and one or more apolipoprotein mimetic peptides. In some embodiments, the complex comprises one or more lipid binding protein molecules (e.g., ApoA-I molecules) instead of one or more apolipoprotein mimetic peptides.

[0136] In some embodiments, the complex comprises 1 to 8 ApoA-I equivalents (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 8, 2 to 6, 2 to 4, 4 to 6, or 4 to 8 ApoA-I equivalents). Lipid binding proteins can be expressed in ApoA-I equivalents depending on the number of amphipathic helices they contain. For example, ApoA-I, which generally exists as a disulfide-bridged dimer, M It can be expressed as 2 ApoA-I equivalents, because each ApoA-I M The molecule contains twice as many amphipathic helices as an ApoA-I molecule. In contrast, a peptide mimetic containing a single amphipathic helix can be expressed as 1 / 10-1 / 6 of the ApoA-I equivalent, since each molecule contains 1 / 10-1 / 6 of the amphipathic helices of an ApoA-I molecule.

[0137] 6.1.5.1. Apolipoprotein

[0138] Suitable apolipoproteins that can be included in the lipid binding protein-based complex include apolipoprotein ApoA-I, ApoA-II, ApoA-IV, ApoA-V, ApoB, ApoC-I, ApoC-II, ApoC-III, ApoD, ApoE, ApoJ, ApoH, and any combination of two or more of the above. Polymorphic forms, isoforms, variants and mutants of the above apolipoproteins, as well as truncated forms, the most common of which is apolipoprotein AI Milano (ApoA-I M ), apolipoprotein AI Paris (ApoA-I P ) and apolipoprotein AI Zaragoza (ApoA-I Z ) can also be used. Apolipoprotein mutants containing cysteine ​​residues are also known and can also be used (see, for example, U.S. Publication No. 2003 / 0181372). Apolipoprotein can be in the form of monomers or dimers, which can be homodimers or heterodimers. For example, ApoA-I (Duverger et al., 1996, Arterioscler. Thromb. Vasc. Biol. 16 (12): 1424-29), ApoA-I M (Franceschini et al., 1985, J. Biol. Chem. 260:1632-35), ApoA-I P(Daum et al., 1999, J. Mol. Med. 77:614-22), ApoA-II (Shelness et al., 1985, J. Biol. Chem. 260(14):8637-46; Shelness et al., 1984, J. Biol. Chem. 259(15):9929-35), ApoA-IV (Duverger et al., 1991, Euro. J. Biochem. 201(2):373-83), ApoE (McLean et al., 1983, J. Biol. Chem. 258(14):8993-9000), homodimers or heterodimers of ApoJ and ApoH (if feasible).

[0139] Apolipoprotein can be modified in its primary sequence to make it less sensitive to oxidation, for example, as described in U.S. Publication Nos. 2008 / 0234192 and 2013 / 0137628 and U.S. Patent Nos. 8,143,224 and 8,541,236. Apolipoprotein can include residues corresponding to elements that facilitate its separation, such as a His tag, or other elements designed for other purposes. Preferably, the apolipoprotein in the complex is soluble in a biological fluid (e.g., lymph, cerebrospinal fluid, vitreous humor, aqueous humor, blood or a blood fraction (e.g., serum or plasma)).

[0140] In some embodiments, the complex comprises covalently bound lipid binding protein monomers, such as dimeric apolipoprotein AI Milano , which is a mutant form of ApoA-I containing cysteine. Cysteine ​​allows the formation of disulfide bonds, which can lead to the formation of homodimers or heterodimers (e.g., ApoA-I Milano-ApoA-II).

[0141] In some embodiments, the apolipoprotein molecule comprises an ApoA-I, ApoA-II, ApoA-IV, ApoA-V, ApoB, ApoC-I, ApoC-II, ApoC-III, ApoD, ApoE, ApoJ, or ApoH molecule, or a combination thereof.

[0142] In some embodiments, the apolipoprotein molecule comprises or consists of an ApoA-I molecule. In some embodiments, the ApoA-I molecule is a human ApoA-I molecule. In some embodiments, the ApoA-I molecule is recombinant. In some embodiments, the ApoA-I molecule is not an ApoA-I. Milano .

[0143] In some embodiments, the ApoA-I molecule is apolipoprotein AIMilano (ApoA-IM), apolipoprotein AI Paris (ApoA-IP) or apolipoprotein AI Zaragoza (ApoA-IZ) molecule.

[0144] Apolipoproteins can be purified from animal sources (particularly from human sources) or recombinantly produced as is well known in the art, see, for example, Chung et al., 1980, J. Lipid Res. 21(3):284-91; Cheung et al., 1987, J. Lipid Res. 28(8):913-29. See also U.S. Pat. Nos. 5,059,528, 5,128,318, 6,617,134; U.S. Publication Nos. 2002 / 0156007, 2004 / 0067873, 2004 / 0077541, and 2004 / 0266660; and PCT Publication Nos. WO 2008 / 104890 and WO 2007 / 023476. Other purification methods are also possible, for example as described in PCT Publication No. WO 2012 / 109162, the disclosure of which is incorporated herein by reference in its entirety.

[0145] Apolipoprotein can be in a pre-pro form, a pro form, or a mature form. For example, the complex can include ApoA-I (e.g., human ApoA-I), wherein the ApoA-I is pre-pro ApoA-I, pro ApoA-I, or mature ApoA-I. In some embodiments, the complex includes ApoA-I having at least 90% sequence identity to SEQ ID NO: 1:

[0146] PPQSPWDRVKDLATVYVDVLKDSGRDYVSQFEGSALGKQLNLKLLDNW

[0147] DSVTSTFSKLREQLGPVTQEFWDNLEKETEGLRQEMSKDLEEVKAKVQP

[0148] YLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEE

[0149] MRDRARAHVDALRTHLAPYSDELRQRLAARLEALKENGGARLAEY(SEQ ID NO:1)

[0150] In other embodiments, the complex comprises an ApoA-I having at least 95% sequence identity to SEQ ID NO: 1. In other embodiments, the complex comprises an ApoA-I having at least 98% sequence identity to SEQ ID NO: 1. In other embodiments, the complex comprises an ApoA-I having at least 99% sequence identity to SEQ ID NO: 1. In other embodiments, the complex comprises an ApoA-I having 100% sequence identity to SEQ ID NO: 1.

[0151] In other embodiments, the complex comprises an ApoA-I having at least 95% sequence identity to amino acids 25 to 267 of SEQ ID NO: 2. In other embodiments, the complex comprises an ApoA-I having at least 98% sequence identity to amino acids 25 to 267 of SEQ ID NO: 2. In other embodiments, the complex comprises an ApoA-I having at least 99% sequence identity to amino acids 25 to 267 of SEQ ID NO: 2. In other embodiments, the complex comprises an ApoA-I having 100% sequence identity to amino acids 25 to 267 of SEQ ID NO: 2.

[0152] In some embodiments, the complex comprises 1 to 8 apolipoprotein molecules (e.g., 1 to 6, 1 to 4, 1 to 2, 2 to 8, 2 to 6, 2 to 4, 4 to 8, 4 to 6, or 6 to 8 apolipoprotein molecules). In some embodiments, the complex comprises 1 apolipoprotein molecule. In some embodiments, the complex comprises 2 apolipoprotein molecules. In some embodiments, the complex comprises 3 apolipoprotein molecules. In some embodiments, the complex comprises 4 apolipoprotein molecules. In some embodiments, the complex comprises 5 apolipoprotein molecules. In some embodiments, the complex comprises 6 apolipoprotein molecules. In some embodiments, the complex comprises 7 apolipoprotein molecules. In some embodiments, the complex comprises 8 apolipoprotein molecules.

[0153] Apolipoprotein molecule can comprise chimeric apolipoprotein, and this chimeric apolipoprotein comprises apolipoprotein and one or more attached functional parts, such as, for example, one or more CRN-001 complexes, one or more targeting moieties, parts with desired biological activity, affinity tags that contribute to purification, and / or reporter molecules for characterization or positioning research. The attached part with biological activity can have the activity that can enhance the biological activity of the compound or cargo part incorporated into the complex of the present disclosure and / or the biological activity of the compound or cargo part incorporated into the complex of the present disclosure and synergizes. For example, the part with biological activity can have antimicrobial (for example, antifungal, antibacterial, antiprotozoal, antibacterial, antifungal or antiviral) activity. In one embodiment, the attached functional part of chimeric apolipoprotein is not in contact with the hydrophobic surface of the complex. In another embodiment, the attached functional part is in contact with the hydrophobic surface of the complex. In some embodiments, the functional part of chimeric apolipoprotein can be inherent to natural protein. In some embodiments, the chimeric apolipoprotein includes a ligand or sequence that is recognized by or is capable of interacting with a cell surface receptor or other cell surface moiety.

[0154] In one embodiment, the chimeric apolipoprotein includes a targeting portion inherent to a non-natural apolipoprotein, such as, for example, S. cerevisiae α-mating factor peptide, folic acid, transferrin or lactoferrin. In another embodiment, the chimeric apolipoprotein includes a portion with a desired biological activity that enhances the activity of the compound or cargo portion incorporated into the complex of the present disclosure and / or synergizes with the activity of the compound or cargo portion incorporated into the complex of the present disclosure. In one embodiment, the chimeric apolipoprotein may include a functional portion inherent to the apolipoprotein. An example of an inherent functional portion of an apolipoprotein is an inherent targeting portion formed approximately by amino acids 130-150 of human ApoE, which includes a receptor binding region recognized by a member of the low-density lipoprotein receptor family. Other examples of inherent functional portions of apolipoproteins include an ApoB-100 region that interacts with a low-density lipoprotein receptor and an ApoA-I region that interacts with a type B1 scavenger receptor. In other embodiments, the functional portion can be added synthetically or recombined to produce a chimeric apolipoprotein. Another example is an apolipoprotein having a prepro or prosequence from another preproapolipoprotein (e.g., a preprosequence from preproapoA-II replaces a preprosequence of preproapoA-I). Another example is an apolipoprotein in which some amphipathic sequence segments have been replaced by other amphipathic sequence segments from another apolipoprotein.

[0155] As used herein, "chimeric" refers to two or more molecules that can exist alone and are connected together to form a single molecule with the desired functions of all its constituent molecules. The constituent molecules of the chimeric molecule can be connected by chemical conjugation synthesis, or when the constituent molecules are all polypeptides or their analogs, the polynucleotides encoding the polypeptides can be recombinantly fused together to express a single continuous polypeptide. Such chimeric molecules are called fusion proteins. "Fusion proteins" are chimeric molecules in which the constituent molecules are all polypeptides and are connected (fused) to each other so that the chimeric molecules form a continuous single chain. The various components can be directly connected to each other or can be coupled by one or more joints. For example, one or more fragments of the various components can be inserted into the sequence of apolipoprotein, or as another example, they can be added to the N-terminus or C-terminus of the sequence of apolipoprotein. For example, the fusion protein can include antibody light chains, antibody fragments, heavy chain antibodies or single domain antibodies.

[0156] In some embodiments, chimeric apolipoprotein is prepared by chemically conjugating apolipoprotein and the functional part to be attached. The method of chemically conjugating molecules is well known to those skilled in the art. Such a mode will vary according to the structure of the part to be attached, but it is easy to determine for those skilled in the art. Polypeptides generally contain a variety of functional groups, such as carboxylic acid (--COOH), free amino groups (--NH2) or sulfhydryl (--SH) groups, which can be used to react with suitable functional groups on the functional part or on the joint to bind the part thereto. The functional part can be attached to the functional group on the N-terminal, C-terminal or internal residue (i.e., the residue at the middle position between the N and C-terminal) of the apolipoprotein molecule. Alternatively, apolipoprotein and / or the part to be tagged can be derivatized to expose or attach other reactive functional groups.

[0157] In some embodiments, fusion proteins including functional portions of polypeptides are synthesized using a recombinant expression system. Typically, this involves creating a nucleic acid (e.g., DNA) sequence encoding apolipoprotein and the functional portion so that the two polypeptides will be in frame when expressed, placing the DNA under the control of a promoter, expressing the protein in a host cell, and isolating the expressed protein.

[0158] The nucleic acid encoding the chimeric apolipoprotein can be incorporated into a recombinant expression vector in a form suitable for expression in a host cell. As used herein, an "expression vector" is a nucleic acid that can be transcribed and translated into a polypeptide when introduced into a suitable host cell. The vector can also include regulatory sequences such as promoters, enhancers or other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are known to those skilled in the art (see, e.g., Goeddel, 1990, Gene Expression Technology: Meth. Enzymol. 185, Academic Press, San Diego, Calif.; Berger and Kimmel, Guide to Molecular Cloning Techniques, Methods in Enzymology 152 Academic Press, Inc., San Diego, Calif.; Sambrook et al., 1989, Molecular Cloning--A Laboratory Manual (2nd ed.) Vol. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor Press, NY, etc.).

[0159] In some embodiments, apolipoprotein has been modified so that when apolipoprotein is incorporated into the complex of the present disclosure, the modification will increase the stability of the complex, confer targeting ability or increase ability. In one embodiment, the modification includes introducing cysteine ​​residues into the apolipoprotein molecule to allow the formation of intramolecular or intermolecular disulfide bonds, such as by site-directed mutagenesis. In another embodiment, a chemical cross-linking agent is used to form intermolecular connections between apolipoprotein molecules to enhance the stability of the complex. Intermolecular cross-linking prevents or reduces the dissociation of apolipoprotein molecules from the complex and / or prevents replacement of endogenous apolipoprotein molecules in a separate body to which the complex is applied. In other embodiments, apolipoprotein is modified by chemical derivatization of one or more amino acid residues or by site-directed mutagenesis to confer cell surface receptor targeting ability or to be recognized by cell surface receptors.

[0160] By engineering receptor recognition properties into apolipoproteins, the complex can be targeted to specific cell surface receptors. For example, the complex can be targeted to a specific cell type known to contain a specific type of infectious agent, for example, by modifying the apolipoprotein so that it can interact with receptors on the surface of the targeted cell type. For example, the complex can be targeted to macrophages by changing the apolipoprotein to confer macrophage endocytic class A scavenger receptor (SR-A) recognition. Modifying the apolipoprotein by site-directed mutagenesis to replace one or more positively charged amino acids with neutral or negatively charged amino acids can confer SR-A binding ability to the complex. SR-A recognition can also be conferred by preparing chimeric apolipoproteins that include N or C-terminal extensions of amino acid sequences with ligands recognized by SR-A or with a high concentration of negatively charged residues. Complexes containing apolipoproteins can also interact with apolipoprotein receptors, such as, but not limited to, ABCA1 receptors, ABCG1 receptors, Megalin, Cubulin, and HDL receptors, such as SR-B1.

[0161] The complex can include a lipid binding protein (e.g., an apolipoprotein molecule) that anchors the cargo portion to the Cargomer. In some embodiments, the apolipoprotein molecule is coupled to the cargo portion by a direct bond. In other embodiments, the apolipoprotein molecule is coupled to the cargo portion by a linker, for example, as described in Section 6.1.8.

[0162] 6.1.5.2. Apolipoprotein mimics

[0163] Peptides, peptide analogs, and agonists that mimic the activity of apolipoprotein (collectively referred to herein as "apolipoprotein peptide mimetics") can also be used alone in the complexes described herein, or in combination with one or more other lipid binding proteins. Peptides and peptide analogs corresponding to apolipoproteins suitable for inclusion in the complexes and compositions described herein, as well as peptides that mimic ApoA-I, ApoA-I MNon-limiting examples of agonists of ApoA-II, ApoA-IV and ApoE activity are disclosed in U.S. Pat. Nos. 6,004,925, 6,037,323 and 6,046,166 to Dasseux et al., U.S. Pat. No. 5,840,688 to Tso, U.S. Pat. No. 6,743,778 to Kohno, U.S. Publication Nos. 2004 / 0266671, 2004 / 0254120, 2003 / 0171277 and 2003 / 0045460 to Fogelman, U.S. Publication No. 2006 / 0069030 to Bachovchin, U.S. Publication No. 2003 / 0087819 to Bielicki, U.S. Publication No. 2009 / 0081293 to Murase et al. al.) and PCT Publication No. WO / 2010 / 093918 (to Dasseux et al.), the disclosures of which are incorporated herein by reference in their entirety. These peptides and peptide analogs can be composed of L-amino acids or D-amino acids or a mixture of L and D amino acids. They can also include one or more non-peptide or amide bonds, such as one or more well-known peptide / amide etc. arrays. Such apolipoprotein peptide mimetics can be synthesized or prepared using any technique known in the art for peptide synthesis, including, for example, the techniques described in U.S. Patent Nos. 6,004,925, 6,037,323, and 6,046,166.

[0164] In some embodiments, the lipid binding protein molecule comprises an apolipoprotein peptide mimetic molecule and optionally one or more apolipoprotein molecules, such as those described above.

[0165] In some embodiments, the apolipoprotein peptidomimetic molecule comprises an ApoA-I peptidomimetic, an ApoA-II peptidomimetic, an ApoA-IV peptidomimetic, or an ApoE peptidomimetic, or a combination thereof.

[0166] The complex of the present disclosure may include an apolipoprotein peptide mimicking molecule that anchors the cargo portion to the complex. In some embodiments, the apolipoprotein peptide mimicking molecule is coupled to the cargo portion by a direct bond. In other embodiments, the apolipoprotein peptide mimicking molecule is coupled to the cargo portion by a linker, for example, as described in Section 6.1.8.

[0167] 6.1.6. Amphiphilic molecules

[0168] Amphiphilic molecules are molecules that have both hydrophobic (non-polar) and hydrophilic (polar) elements. Amphiphilic molecules that can be used in the complexes described herein include lipids (e.g., as described in Section 6.1.6.1), detergents (e.g., as described in Section 6.1.6.2), fatty acids (e.g., as described in Section 6.1.6.3), and non-polar molecules covalently attached to polar molecules (such as, but not limited to, sugars or nucleic acids) and sterols (e.g., as described in Section 6.1.6.4).

[0169] The complex can include a single type of amphiphilic molecule (e.g., a single type of phospholipid or a mixture of phospholipids) or can contain a combination of multiple types of amphiphilic molecules (e.g., a phospholipid and a detergent). The complex can contain an amphiphilic molecule or a combination of amphiphilic molecules that are configured to promote the solubilization of the lipid binding protein molecule.

[0170] In some embodiments, the apomer and / or cargomer-based complexes contain only amphiphilic molecules in an amount sufficient to solubilize the lipid-binding protein molecule. In other words, the apomer and / or cargomer-based complexes can contain the minimum amount of one or more amphiphilic molecules necessary to solubilize the lipid-binding protein molecule.

[0171] In some embodiments, the amphiphilic molecules included comprise a phospholipid, a detergent, a fatty acid, a non-polar moiety covalently linked to a sugar or a sterol, or a combination thereof (eg, selected from the types of amphiphilic molecules discussed above).

[0172] In some embodiments, the amphiphilic molecule comprises or is composed of a phospholipid molecule. In some embodiments, the phospholipid molecule comprises a negatively charged phospholipid, a neutral phospholipid, a positively charged phospholipid or a combination thereof. In some embodiments, the phospholipid molecule contributes 1-3 net charges to each apolipoprotein molecule in the complex. In some embodiments, the net charge is a negative net charge. In some embodiments, the net charge is a positive net charge. In some embodiments, the phospholipid molecule is composed of a combination of negatively charged phospholipids and neutral phospholipids. In some embodiments, the molar ratio of negatively charged phospholipids to neutral phospholipids ranges from 1:1 to 1:3, for example, about 1:1, about 1:2 or about 1:3. In some embodiments, the molar ratio of negatively charged phospholipids to neutral phospholipids is about 1:1 or about 1:2. In some embodiments, the weight ratio of neutral phospholipids to negatively charged phospholipids ranges from 95:5 to 99:1.

[0173] In some embodiments, the complex comprises at least one amphiphilic molecule that serves as an anchor.

[0174] In some embodiments, the amphiphilic molecule comprises a neutral phospholipid and a negatively charged phospholipid in a weight ratio of 95:5 to 99:1.

[0175] 6.1.6.1. Lipids

[0176] The complex based on lipid-binding protein can include one or more lipids. In various embodiments, one or more lipids can be saturated and / or unsaturated, natural and / or synthetic, charged or uncharged, zwitterionic or non-zwitterionic. In some embodiments, lipid molecules (e.g., phospholipid molecules) can contribute 1-3 (e.g., 1-3, 1-2, 2-3, 1, 2 or 3) net charges to each lipid-binding protein molecule in the complex together. In some embodiments, the net charge is negative. In other embodiments, the net charge is positive.

[0177] In some embodiments, the lipid comprises a phospholipid. The phospholipid can have two identical or different acyl chains (e.g., chains with different numbers of carbon atoms, different saturations between acyl chains, different branching of acyl chains, or a combination thereof). The lipid can also be modified to contain a fluorescent probe (e.g., as described in avantilipids.com / product-category / products / fluorescent-lipids / ). Preferably, the lipid comprises at least one phospholipid.

[0178] The phospholipids can have unsaturated or saturated acyl chains of about 6 to about 24 carbon atoms (e.g., 6-20, 6-16, 6-12, 12-24, 12-20, 12-16, 16-24, 16-20, or 20-24). In some embodiments, the phospholipids used in the complexes of the present disclosure have one or two acyl chains of 12, 14, 16, 18, 20, 22, or 24 carbon atoms (e.g., two acyl chains of the same length or two acyl chains of different lengths).

[0179] Non-limiting examples of acyl chains present in common fatty acids that may be contained in phospholipids are as follows:

[0180] Table 1 provides:

[0181]

[0182] The lipids that may be present in the complexes of the present disclosure include, but are not limited to, small alkyl chain phospholipids, egg phosphatidylcholine, soybean phosphatidylcholine, dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, distearoylphosphatidylcholine, 1-myristoyl-2-palmitoylphosphatidylcholine, 1-palmitoyl-2-myristoylphosphatidylcholine, 1-palmitoyl-2-stearoylphosphatidylcholine, 1-stearoyl-2-palmitoylphosphatidylcholine, dioleoylphosphatidylcholine, dilauroylphosphatidylglycerol, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol (e.g., dimyristoylphosphatidylglycerol), dipalmitoylphosphatidylglycerol Oil, distearoylphosphatidylglycerol, dioleoylphosphatidylglycerol, dimyristoylphosphatidic acid, dipalmitoylphosphatidic acid, dimyristoylphosphatidylamine, dipalmitoylphosphatidylamine, dimyristoylphosphatidylserine, dipalmitoylphosphatidylserine, brain phosphatidylserine, brain sphingomyelin, palmitoyl sphingomyelin, dipalmitoyl sphingomyelin, egg sphingomyelin, milk sphingomyelin, plant sphingomyelin, distearoyl sphingomyelin, dipalmitoylphosphatidylglycerol salt, phosphatidic acid, galactoside, ganglioside, cerebroside, dilaurylphosphatidylcholine, (1,3)-D-mannosyl-(1,3) diacylglycerol, aminophenyl glycoside, 3-cholesterol-6'-(glycosylthio)hexyl ether glycolipid, cholesterol and its derivatives. Synthetic lipids such as synthetic palmitoyl sphingomyelin or N-palmitoyl-4-hydroxysphingosine-1-phosphocholine (a form of plant sphingomyelin) can be used to minimize lipid oxidation.

[0183] In some embodiments, the lipid-binding protein-based complex includes two types of phospholipids: neutral lipids, such as phosphatidylcholine and / or sphingomyelin (abbreviated as SM), and charged phospholipids (e.g., negatively charged phospholipids). "Neutral" phospholipids have a net charge of about zero at physiological pH. In many embodiments, neutral phospholipids are zwitterions, although other types of net neutral phospholipids are known and can be used. In some embodiments, the molar ratio of charged phospholipids (e.g., negatively charged phospholipids) to neutral phospholipids ranges from 1:1 to 1:3, such as about 1:1, about 1:2, or about 1:3.

[0184] Neutral phospholipids can include, for example, one or both of lecithin and / or SM, and can optionally include other neutral phospholipids. In some embodiments, neutral phospholipids include lecithin, but do not include SM. In other embodiments, neutral phospholipids include SM, but do not include lecithin. In other embodiments, neutral phospholipids include lecithin and SM. Except lecithin and / or SM, all these specific illustrative examples can include neutral phospholipids, but do not include this type of other neutral phospholipids in many embodiments.

[0185] As used herein, the expression of "SM" includes sphingomyelin derived from or obtained from natural sources, and analogs and derivatives of naturally occurring SM, which are not affected by LCAT hydrolysis, as is the case with naturally occurring SM.SM is a phospholipid that is very similar to lecithin in structure, but unlike lecithin, it does not have a glycerol backbone, and therefore does not have the ester bond of an acyl chain attached.On the contrary, SM has a ceramide backbone, and amide bonds connect acyl chains.SM can be obtained from, for example, milk, eggs or brain.SM analogs or derivatives can also be used. The non-limiting examples of useful SM analogs and derivatives include but are not limited to palmitoyl sphingomyelin, N-palmitoyl-4-hydroxysphingosine-1-phosphocholine (a form of plant sphingomyelin), palmitoyl sphingomyelin, stearoyl sphingomyelin, D-erythro-N-16:0-sphingomyelin and its dihydro isomer, D-erythro-N-16:0-dihydro-sphingomyelin. Synthetic SM, such as synthetic palmitoyl sphingomyelin or N-palmitoyl-4-hydroxysphingosine-1-phosphocholine (phytosphingomyelin), can be used to produce more homogeneous complexes and fewer contaminants and / or oxidation products than sphingolipids from animal sources. Methods for synthesizing SM are described in U.S. Publication No. 2016 / 0075634.

[0186] Sphingomyelin isolated from natural sources can be artificially enriched with a specific saturated or unsaturated acyl chain. For example, milk sphingomyelin (Avanti Phospholipid, Alabaster, Ala.) is characterized by long saturated acyl chains (i.e., acyl chains with 20 or more carbon atoms). In contrast, egg sphingomyelin is characterized by short saturated acyl chains (i.e., acyl chains with less than 20 carbon atoms). For example, while only about 20% of milk sphingomyelin contains C16:0 (16 carbons, saturated) acyl chains, about 80% of egg sphingomyelin contains C16:0 acyl chains. Using solvent extraction, the composition of milk sphingomyelin can be enriched to have an acyl chain composition comparable to egg sphingomyelin, and vice versa.

[0187] SM can be semi-synthetic, so it has a specific acyl chain. For example, milk sphingomyelin can first be purified from milk, and then a specific acyl chain, such as a C16:0 acyl chain, can be cleaved and replaced by another acyl chain. SM can also be completely synthesized, for example by large-scale synthesis. See, for example, U.S. Patent Application No. 5,220,043 entitled Synthesis of D-erythro-sphingomyelin published by Dong et al. on June 15, 1993; Weis, 1999, Chem.Phys.Lipids 102(1-2):3-12. SM can be completely synthesized, for example, as described in U.S. Publication No. 2014 / 0275590.

[0188] The length and saturation level of the acyl chain comprising a semisynthetic or synthetic SM can be selectively changed. The acyl chain can be saturated or unsaturated, and can contain about 6 to about 24 carbon atoms. Each chain can contain the same number of carbon atoms, or each chain can contain different numbers of carbon atoms. In some embodiments, a semisynthetic or synthetic SM comprises a mixed acyl chain so that one chain is saturated and one chain is unsaturated. In such mixed acyl chain SMs, the chain length can be the same or different. In other embodiments, the acyl chain of a semisynthetic or synthetic SM is either saturated or unsaturated. Equally, the chain can contain the same or different numbers of carbon atoms. In some embodiments, two acyl chains comprising a semisynthetic or synthetic SM are identical. In a specific embodiment, these chains correspond to the acyl chains of naturally occurring fatty acids, such as, for example, oleic acid, palmitic acid or stearic acid. In another embodiment, a SM with a saturated or unsaturated functionalized chain is used. In another specific embodiment, two acyl chains are all saturated and contain 6 to 24 carbon atoms. Non-limiting examples of acyl chains present in common fatty acids that may be included in semi-synthetic and synthetic SMs are provided in Table 1 above.

[0189] In some embodiments, the SM is a palmitoyl SM, such as a synthetic palmitoyl SM, which has a C16:0 acyl chain, or an egg SM, which includes a palmitoyl SM as a major component.

[0190] In specific embodiments, a functionalized SM is used, such as a phytosphingomyelin.

[0191] Lecithin can be derived from natural sources or separated, or can be synthesized. The example of suitable lecithin separated from natural sources includes but is not limited to egg phosphatidylcholine and soybean phosphatidylcholine. The other non-restrictive example of suitable lecithin comprises dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, distearoylphosphatidylcholine 1-myristoyl 1-2-palmitoylphosphatidylcholine, 1-palmitoyl 1-2-myristoylphosphatidylcholine, 1-palmitoyl 1-2-stearoylphosphatidylcholine, 1-stearoyl 1-2-palmitoylphosphatidylcholine, 1-palmitoyl 1-2-oleoylphosphatidylcholine, 1-stearoyl 1-2-palmitoylphosphatidylcholine, dioleoylphosphatidylcholine and its ether derivatives or analogs.

[0192] The lecithin derived from or separated by natural sources can be enriched to include specific acyl chains. In the embodiment of semisynthetic or synthetic lecithin, the characteristics of the acyl chains can be selectively changed, as discussed above in conjunction with SM. In some embodiments of complex as described herein, two acyl chains on the lecithin are identical. In some embodiments of the complex comprising SM and lecithin, the acyl chains of SM and lecithin are all identical. In a specific embodiment, the acyl chains correspond to myristic acid, palmitic acid, oleic acid or stearic acyl chains.

[0193] The complex of the present disclosure may include one or more negatively charged phospholipids (e.g., alone or in combination with one or more neutral phospholipids). As used herein, "negatively charged phospholipids" are phospholipids with a net negative charge at physiological pH. Negatively charged phospholipids may include a single type of negatively charged phospholipids, or a mixture of two or more different negatively charged phospholipids. In some embodiments, the charged phospholipids are negatively charged glycerophospholipids. Specific examples of suitable negatively charged phospholipids include, but are not limited to, 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)], phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, phosphatidic acid and salts thereof (e.g., sodium or potassium salts). In some embodiments, negatively charged phospholipids include one or more of phosphatidylinositol, phosphatidylserine, phosphatidylglycerol and / or phosphatidic acid. In a specific embodiment, the negatively charged phospholipids include or consist of a salt of phosphatidylglycerol or a salt of phosphatidylinositol. In another specific embodiment, the negatively charged phospholipid comprises or consists of 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] or DPPG or a salt thereof.

[0194] Negatively charged phospholipids can be obtained from natural sources or prepared by chemical synthesis. In embodiments using synthetic negatively charged phospholipids, the characteristics of the acyl chains can be selectively changed, as discussed above in conjunction with SM. In some embodiments of the complex of the present invention, the two acyl chains on the negatively charged phospholipids are the same. In some embodiments, the acyl chains of all types of phospholipids included in the complex of the present invention are the same. In a specific embodiment, the complex comprises negatively charged phospholipids and / or SM, all of which have C16:0 or C16:1 acyl chains. In a specific embodiment, the fatty acid portion of SM is mainly C16:1 palmitoyl. In a specific embodiment, the acyl chains of charged phospholipids, phosphatidylcholine and / or SM correspond to the acyl chains of palmitic acid. In another specific embodiment, the acyl chains of charged phospholipids, phosphatidylcholine and / or SM correspond to the acyl chains of oleic acid.

[0195] Examples of positively charged phospholipids that can be included in the complexes of the present disclosure include N1-[2-((1S)-1-[(3-aminopropyl)amino]-4-[di(3-amino-propyl)amino]butylcarboxamido)ethyl]-3,4-di[oleyloxy]-benzamide, 1,2-di-O-octadecenyl-3-trimethylpropane ammonium, 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine, 1-palmitoyl-2- Oleoyl-sn-glycero-3-ethylphosphocholine, 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine, 1,2-distearoyl-sn-glycero-3-ethylphosphocholine, 1,2-dipalmitoyl-sn-glycero-3-ethylphosphocholine, 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine, 1,2-dilauroyl-sn-glycero-3-ethylphosphocholine, 1,2-dilauroyl-sn-glycero -3-ethylphosphocholine, 1,2-dioleoyl-3-dimethylammonium-propane 1,2-dimyristoyl-3-dimethylammonium-propane, 1,2-dipalmitoyl-3-dimethylammonium-propane, N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleoyloxy))propane-1-amine, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-stearoyl-3- Trimethylammonium-propane, 1,2-dipalmitoyl-3-trimethylammonium-propane, 1,2-dimyristoyl-3-trimethylammonium-propane, N-[1-(2,3-dimyristyloxy)propyl]-N,N-dimethyl-N-(2-hydroxyethyl)ammonium bromide, N,N,N-trimethyl-2-bis[(1-oxo-9-octadecenyl)oxy]-(Z,Z)-1-propylamine methylsulfate and salts thereof (e.g., chloride or bromide salts).

[0196] The lipids used are preferably at least 95% pure and / or have a reduced level of oxidants (such as, but not limited to, peroxides). Lipids obtained from natural sources preferably have less polyunsaturated fatty acid moieties and / or less oxidizable fatty acid moieties. The oxidation level in a sample can be determined using iodine titration, which provides a peroxide value, expressed as milliequivalents of iodine separated per kilogram of sample, abbreviated as meq O / kg. See, e.g., Gray, 1978, Measurement of Lipid Oxidation: A Review, Journal of the American Oil Chemists Society 55: 539-545; Heaton, FW and Ur, Improved Iodometric Methods for the Determination of Lipid Peroxides, 1958, Journal of the Science of Food and Agriculture 9: 781-786. Preferably, the oxidation level or peroxide level is low, e.g., less than 5 meq O / kg, less than 4 meq O / kg, less than 3 meq O / kg, or less than 2 meq O / kg.

[0197] In some embodiments, the complex may include a small amount of additional lipids. Virtually any type of lipid may be used, including but not limited to lysophospholipids, galactocerebrosides, gangliosides, cerebrosides, glycerides, triglycerides, and sterols and sterol derivatives (e.g., phytosterols, animal sterols, such as cholesterol, or sterol derivatives, such as cholesterol derivatives). For example, the complex of the present invention may contain cholesterol or a cholesterol derivative, such as a cholesterol ester. The cholesterol derivative may also be a substituted cholesterol or a substituted cholesterol ester. The complex of the present invention may also contain oxidized sterols, such as but not limited to oxidized cholesterol or oxidized sterol derivatives (such as but not limited to oxidized cholesterol esters). In some embodiments, the complex does not include cholesterol and / or its derivatives (such as cholesterol esters or oxidized cholesterol esters).

[0198] 6.1.6.2. Detergents

[0199] The complex may contain one or more detergents. The detergent may be zwitterionic, nonionic, cationic, anionic, or a combination thereof. Exemplary zwitterionic detergents include 3-[(3-cholamidopropyl)dimethylamino]-1-propanesulfonate (CHAPS), 3-[(3-cholamidopropyl)dimethylamino]-2-hydroxy-1-propanesulfonate (CHAPSO), and N,N-dimethyldodecylamine N-oxide (LDAO). Exemplary nonionic detergents include D-(+)-trehalose 6-monooleate, N-octanoyl-N-methylglucamine, N-nonanoyl-N-methylglucamine, N-decanoyl-N-methylglucamine, 1-(7Z-hexadecenoyl)-rac-glycerol, 1-(8Z-hexadecenoyl)-rac-glycerol, 1-(8Z-heptadecenoyl)-rac-glycerol, 1-(9Z-hexadecenoyl)-rac-glycerol, 1-decanoyl-rac-glycerol. Exemplary cationic detergents include (S)-O-methyl-serine dodecylamide hydrochloride, dodecyl ammonium chloride, decyltrimethylammonium bromide and hexadecyltrimethylammonium sulfate. Exemplary anionic detergents include cholesterol hemisuccinate, cholate, alkyl sulfates and alkyl sulfonates.

[0200] 6.1.6.3. Fatty acids

[0201] The complex can contain one or more fatty acids. The one or more fatty acids can include short-chain fatty acids (e.g., butyric acid, isobutyric acid, valeric acid or isovaleric acid) with five or less aliphatic tails of carbon, medium-chain fatty acids (e.g., caproic acid, caprylic acid, capric acid or lauric acid) with aliphatic tails of 6 to 12 carbons, long-chain fatty acids (e.g., myristic acid, palmitic acid, stearic acid or arachidic acid) with aliphatic tails of 13 to 21 carbons, very long-chain fatty acids (e.g., behenic acid, lignoceric acid or cerotic acid) with aliphatic tails of 22 or more carbons, or combinations thereof. The one or more fatty acids can be saturated (e.g., caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, eicosanoic acid, or cerotic acid), unsaturated (e.g., myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, or docosahexaenoic acid), or a combination thereof. The unsaturated fatty acids can be cis or trans fatty acids. In some embodiments, the unsaturated fatty acids used in the complexes of the present disclosure are cis fatty acids.

[0202] 6.1.6.4. Nonpolar molecules and sterols attached to sugars

[0203] The complex may contain one or more amphiphilic molecules comprising a non-polar molecule or moiety (e.g., a hydrocarbon chain, an acyl or diacyl chain) or a sterol (e.g., cholesterol) linked to a sugar (e.g., a monosaccharide such as glucose or galactose, or a disaccharide such as maltose or trehalose). The sugar may be a modified sugar or a substituted sugar. Exemplary amphiphilic molecules comprising a non-polar molecule linked to a sugar include dodec-2-yloxy-β-D-maltoside, tridec-3-yloxy-β-D-maltoside, tridec-2-yloxy-β-D-maltoside, n-dodecyl-β-D-maltoside (DDM), n-octyl-β-D-glucoside, n-nonyl-β-D-glucoside, n-decyl-β-D-maltoside, n-dodecyl-β-D-maltopyranoside, 4-n-dodecyl-α,α-trehalose, 6-n-dodecyl-α,α-trehalose, and 3-n-dodecyl-α,α-trehalose.

[0204] In some embodiments, the polar moiety is an acyl or diacyl chain.

[0205] In some embodiments, the sugar is a modified sugar or a substituted sugar.

[0206] 6.1.7. Anchor

[0207] The cargo moiety can be covalently bound to an amphiphilic or non-polar moiety to facilitate the coupling of the cargo moiety to the lipid-binding protein-based complex. The amphiphilic and non-polar moieties can interact with the non-polar regions in the lipid-binding protein-based complex, thereby anchoring the cargo moiety attached to the amphiphilic and non-polar moieties to the complex.

[0208] Amphiphilic moieties that can be used as anchors include lipids (e.g., as described in Section 6.1.6.1) and fatty acids (e.g., as described in Section 6.1.6.3). In some embodiments, the anchor comprises a sterol or a sterol derivative, such as a plant sterol, an animal sterol, or a sterol derivative such as a vitamin. For example, a sterol such as cholesterol can be covalently bound to a cargo moiety (e.g., via the hydroxyl group at position 3 of the sterol A ring) and used to anchor the cargo moiety to the complex. Non-polar moieties that can be used as anchors include alkyl chains, acyl chains, and diacyl chains. The cargo moiety can be covalently bound to the anchor moiety directly or indirectly via a linker (e.g., by a bifunctional peptide or other linkers described in Section 6.1.8). A cargo moiety with biological activity can retain its biological activity when covalently bound to an anchor (or a linker attached to an anchor), while others may require cleavage of the covalent bond (e.g., by hydrolysis) to attach the cargo moiety to the anchor (or a linker attached to an anchor) to restore biological activity.

[0209] In some embodiments, at least one cargo moiety is coupled to an anchor. In some embodiments, the anchor comprises an amphiphilic and / or non-polar moiety. In some embodiments, the anchor comprises an amphiphilic moiety. In some embodiments, the amphiphilic moiety comprises an amphiphilic molecule in the complex. In some embodiments, the amphiphilic moiety comprises a lipid, a detergent, a fatty acid, a non-polar molecule attached to a sugar, or a sterol attached to a sugar.

[0210] In some embodiments, the amphiphilic moiety comprises a sterol. In some embodiments, the sterol comprises an animal sterol or a plant sterol. In some embodiments, the sterol comprises cholesterol.

[0211] In other embodiments, the anchor comprises a non-polar moiety. In some embodiments, the non-polar moiety comprises an alkyl chain, an acyl chain, or a diacyl chain.

[0212] In some embodiments, the cargo moiety is coupled to the anchor via a direct bond.

[0213] In some embodiments, the cargo moiety is coupled to the anchor via a linker.

[0214] 6.1.8. Connectors

[0215] The linker includes a chain of atoms that covalently links the cargo moiety to other parts in the cargo complex such as a cargomer, such as an apolipoprotein molecule, an amphiphilic molecule, and an anchor. Many linker molecules are commercially available, such as from ThermoFisherScientific. Suitable linkers are well known to those skilled in the art, and include but are not limited to straight or branched carbon linkers, heterocyclic carbon linkers, and peptide linkers. The linker can be a bifunctional linker, which is homobifunctional or heterobifunctional.

[0216] Suitable linkers include cleavable and non-cleavable linkers.

[0217] The linker can be a cleavable linker to promote the release of the cargo portion in vivo. Cleaving linkers include acid-labile linkers (e.g., containing hydrazine or cis-aconyl), protease-sensitive (e.g., peptidase-sensitive) linkers, photolabile linkers, or disulfide-containing linkers (Chari et al., 1992, Cancer Research 52: 127-131; U.S. Patent No. 5,208,020). Cleaving linkers are generally easy to cleave under intracellular conditions. Suitable cleavable linkers include, for example, peptide linkers that can be cleaved by intracellular proteases, such as lysosomal proteases or endosomal proteases. In an exemplary embodiment, the linker can be a dipeptide linker, such as a valine-citrulline (val-cit) or a phenylalanine-lysine (phe-lys) linker.

[0218] Cleavable linkers can be pH sensitive, i.e., sensitive to hydrolysis at certain pH values. Typically, pH sensitive linkers are hydrolyzable under acidic conditions. For example, acid-labile linkers (e.g., hydrazones, semicarbazones, thiosemicarbazones, cis-aconitamides, orthoesters, acetals, ketals, etc.) that are hydrolyzable in lysosomes can be used. (See, e.g., U.S. Patent Nos. 5,122,368; 5,824,805; 5,622,929; Dubowchik and Walker, 1999, Pharm. Therapeutics 83: 67-123; Neville et al., 1989, Biol. Chem. 264: 14653-14661). Such linkers are relatively stable under neutral pH conditions, such as those in blood, but are unstable when they are below pH 5.5 or 5.0 (approximate lysosomal pH). In certain embodiments, the hydrolyzable linker is a thioether linker (e.g., a thioether linked to the cargo moiety via an acylhydrazone bond (see, e.g., U.S. Pat. No. 5,622,929).

[0219] In some embodiments, the linker is cleavable under reducing conditions (eg, a disulfide linker). A variety of disulfide linkers are known in the art, including, for example, those that can be formed using SATA (N-succinimidyl-5-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio) propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio) butyrate), and SMPT (N-succinimidyloxycarbonyl-α-methyl-α-(2-pyridyl-dithio) toluene), SPDB and SMPT (see, e.g., Thorpe et al., 1987, Cancer Res. 47:5924-5931; Wawrzynczak et al., In Immunoconjugates: Antibody Conjugates in Radioimagery and Therapy of Cancer (CW Vogeled., Oxford U. Press, 1987. See also U.S. Pat. No. 4,880,935).

[0220] In some embodiments, the joint can be cleaved by a cleavage agent (e.g., enzyme) present in the intracellular environment (e.g., lysosomes or endosomes or caveolae). The joint can be, for example, a peptidyl joint cleaved by an intracellular peptidase or protease, including but not limited to lysosomal or endosomal proteases. In some embodiments, the peptidyl joint is at least two amino acids long or at least three amino acids long. The cleavage agent can include cathepsins B and D and plasmin, all of which are known to hydrolyze dipeptide drug derivatives, resulting in the release of active drugs in target cells (see, e.g., Dubowchik and Walker, 1999, Pharm. Therapeutics 83: 67-123). In some embodiments, the peptidyl joint that can be cleaved by intracellular proteases is a Val-Cit joint or a Phe-Lys joint.

[0221] In some embodiments, the linker is a malonate linker (Johnson et al., 1995, Anticancer Res. 15: 1387-93), a maleimidobenzoyl linker (Lau et al., 1995, Bioorg-Med-Chem. 3(10): 1299-1304), or a 3'-N-amide analog (Lau et al., 1995, Bioorg-Med-Chem. 3(10): 1305-12).

[0222] In other embodiments, the linker unit is not cleavable and the cargo moiety is released, for example, by degradation of the complex. Exemplary non-cleavable linkers include maleimidocaproyl, N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), and N-succinimidyl-4-(iodoacetyl)-aminobenzoate (SIAB).

[0223] In some embodiments, the cargo portion is coupled to the anchor via a linker (e.g., as described in Section 6.1.7). In some embodiments, the linker coupling the cargo portion to the anchor is a bifunctional linker. In some embodiments, the linker coupling the cargo portion to the anchor is a cleavable linker. In some embodiments, the cleavable linker is a dipeptide linker such as a valine-citrulline (val-cit) or a phenylalanine-lysine (phe-lys) linker. In some embodiments, the linker coupling the cargo portion to the anchor is a non-cleavable linker. Exemplary non-cleavable linkers include maleimidocaproyl, N-succinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (SMCC) and N-succinimidyl-4-(iodoacetyl)-aminobenzoate (SIAB).

[0224] 6.1.9. Ophthalmic drugs

[0225] In some embodiments of the methods described herein, a lipid-binding protein-based complex (e.g., CER-001) can be used as a carrier to deliver one or more ophthalmic drugs to the eyes of a subject. In some embodiments, one or more ophthalmic drugs can be considered as a cargo portion and can be non-covalently or covalently compounded to a component of the complex (e.g., by an anchor or a linker) compounded to a lipid-binding protein-based complex (e.g., CER-001). In some embodiments, one or more ophthalmic drugs are not covalently attached to the complex. One or more ophthalmic drugs can be added to a preformed complex, such as a preformed CER-001, to prepare a complex further comprising one or more ophthalmic drugs. One or more ophthalmic drugs can be promoted by performing one or more heating and cooling cycles to compound with the preformed complex, for example, as described in Example 1. Alternatively, one or more ophthalmic drugs can be included in the process for preparing the complex, for example, in a starting suspension comprising lipid-binding proteins and lipid components that undergoes thermal cycling. The thermal cycling process for preparing a lipid-binding protein-based complex is described in WO 2012 / 109162 and WO / 2019 / 030574.

[0226] In some embodiments, the one or more ophthalmic drugs include steroids, kinase inhibitors, angiotensin II receptor antagonists, aldose reductase inhibitors, immunosuppressants, carbonic anhydrase inhibitors, antimicrobial agents, antiviral agents, antihistamines, anti-inflammatory agents, prostaglandin analogs, or combinations thereof.

[0227] Exemplary ophthalmic drugs that can be used include, but are not limited to, steroids such as dexamethasone, dexamethasone palmitate, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, and spironolactone; kinase inhibitors such as axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)- 4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafil angiotensin II receptor antagonists such as candesartan, irbesartan, losartan, olmesartan, telmisartan, and valsartan; aldose reductase inhibitors such as 2-methylsorbitol; immunosuppressants such as sirolimus, cyclosporine, and tacrolimus; carbonic anhydrase inhibitors such as acetazolamide, brinzolamide, and dorzoxan; antimicrobial, antifungal, and antiviral agents, such as azithromycin, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, and zidovudine; antihistamines, such as levocabastine; nonsteroidal anti-inflammatory drugs, such as bromfenac, diclofenac, indomethacin, and nepafenac; and prostaglandin analogs, such as latanoprost, trovaprost, bimatoprost, and tafluprost.

[0228] In some embodiments, the lipid-binding protein-based complex comprises a prostaglandin analog, such as latanoprost, trovaprost, bimatoprost, tafluprost, or a combination thereof. In a specific embodiment, the lipid-binding protein-based complex comprises latanoprost. In another embodiment, the lipid-binding protein-based complex comprises trovaprost. In another embodiment, the lipid-binding protein-based complex comprises bimatoprost. In another embodiment, the lipid-binding protein-based complex comprises tafluprost.

[0229] In some embodiments, the lipid binding protein-based complex comprises dexamethasone, axitinib, cediranib, dovitinib, motesanib, pazopanib, regorafenib, losartan, olmesartan, dorzolamide, diclofenac, nepafenac, or a combination thereof.

[0230] In other embodiments, the lipid binding protein-based complex includes azithromycin.

[0231] In other embodiments, the lipid binding protein-based complex comprises spironolactone.

[0232] In other embodiments, the lipid binding protein-based complex comprises dexamethasone palmitate.

[0233] In other embodiments, the lipid binding protein-based complex comprises cyclosporine.

[0234] In other embodiments, the lipid binding protein-based complex comprises dexamethasone.

[0235] In other embodiments, the lipid binding protein-based complex comprises loteprednol etabonate.

[0236] In other embodiments, the lipid binding protein-based complex comprises triamcinolone acetonide.

[0237] In other embodiments, the lipid binding protein-based complex comprises acyclovir.

[0238] In other embodiments, the lipid binding protein-based complex comprises pazopanib.

[0239] In other embodiments, the lipid binding protein-based complex comprises sirolimus.

[0240] In other embodiments, the lipid binding protein-based complex comprises tacrolimus.

[0241] In other embodiments, the lipid binding protein-based complex comprises nepafenac.

[0242] 6.1.9.1. Combination of CER-001 and ophthalmic drugs

[0243] As disclosed herein, CER-001 can be used as a drug carrier to deliver one or more ophthalmic drugs to the eye of a subject. Accordingly, the present disclosure provides a composition comprising CER-001 and one or more ophthalmic drugs (e.g., one or more hydrophobic and / or poorly water soluble or insoluble drugs).

[0244] In one embodiment, the composition comprises CER-001 and a steroid. In some embodiments, the composition comprises CER-001 and dexamethasone. In some embodiments, the composition comprises CER-001 and dexamethasone palmitate. In some embodiments, the composition comprises CER-001 and loteprednol etabonate. In some embodiments, the composition comprises CER-001 and triamcinolone acetonide.

[0245] In another embodiment, the composition comprises CER-001 and an antimicrobial, antifungal, or antiviral agent. In some embodiments, the composition comprises CER-001 and azithromycin. In some embodiments, the composition comprises CER-001 and acyclovir.

[0246] In another embodiment, the composition comprises CER-001 and a prostaglandin analog. In some embodiments, the composition comprises CER-001 and latanoprost. In some embodiments, the composition comprises CER-001 and trovaprost. In some embodiments, the composition comprises CER-001 and bimatoprost. In some embodiments, the composition comprises CER-001 and tafluprost.

[0247] In another embodiment, the composition comprises CER-001 and a kinase inhibitor. In some embodiments, the composition comprises CER-001 and pazopanib.

[0248] In another embodiment, the composition comprises CER-001 and an immunosuppressant. In some embodiments, the composition comprises CER-001 and sirolimus. In some embodiments, the composition comprises CER-001 and tacrolimus.

[0249] In another embodiment, the composition comprises CER-001 and a nonsteroidal anti-inflammatory drug. In some embodiments, the composition comprises CER-001 and nepafenac.

[0250] In a further embodiment, the composition comprises CER-001 and spironolactone.

[0251] In another embodiment, the composition comprises CER-001 and cyclosporine.

[0252] The composition described in this section 6.1.9.1 can be prepared by any suitable means, for example, as described in Section 6.1.10, for example, by thermal cycling a mixture containing CER-001 and an ophthalmic drug. The composition can be appropriately formulated for the intended route of administration, such as topical administration, such as topical administration or intraocular administration. The composition for intraocular administration can be formulated to be administered by, for example, intraocular injection, such as intravitreal injection, subconjunctival injection, juxtabular injection, peribulbar injection, retrobulbar injection, suprachoroidal injection or suprascleral injection. For topical administration, the composition can be formulated for administration, for example, as eye drops. The composition can also be formulated as an implant, formulated for administration via iontophoresis, or formulated for administration via electroporation. In some embodiments, the composition is formulated for suprachoroidal injection. In other embodiments, the composition is formulated for injection on the sclera. In other embodiments, the composition is formulated as an implant (e.g., a disc, a sheet, a plug, a rod or a pellet). In other embodiments, the composition is formulated for administration via iontophoresis. In other embodiments, the composition is formulated for administration via electroporation.

[0253] 6.1.10. Preparation

[0254] The lipid-binding protein-based complex can be formulated for the intended route of administration, for example, according to techniques known in the art (e.g., as described in Allen et al., eds., 2012, Remington: The Science and Practice of Pharmacy, 22nd Edition, Pharmaceutical Press, London, UK; or as described in Davis et al., 2004, Curr Opin Mol Ther. 6(2): 195-205). In some embodiments, the formulation comprises a lipid-binding protein-based complex, such as CER-001, and one or more ophthalmic drugs, such as one or more ophthalmic drugs described in Section 6.1.9.

[0255] In some embodiments, the lipid binding protein-based complex, such as CER-001, and optionally one or more ophthalmic drugs, is formulated for delivery via intraocular injection, such as intravitreal injection, subconjunctival injection, juxtabulbar injection, peribulbar injection, retrobulbar injection, suprachoroidal injection, or suprascleral injection.

[0256] In some embodiments, the formulation is formulated for suprachoroidal injection. Suprachoroidal injection can be used to achieve higher chorioretinal drug concentrations than achieved by traditional intravitreal injection. In other embodiments, the formulation is formulated for supra-scleral injection.

[0257] In some embodiments, a lipid binding protein-based complex, such as CER-001 and optionally one or more ophthalmic drugs, is formulated for delivery as an implant. Implants disclosed herein can have various shapes, such as discs, sheets, plugs, rods, or pellets. Implants can be biodegradable. Biodegradable implants can be formulated with materials such as polylactic acid and / or polylactic-glycolic acid, which degrade over time. Alternatively, implants can also be non-biodegradable. Non-biodegradable implants can be formulated with materials such as silicon or polymers such as ethylene vinyl acetate or polyvinyl alcohol.

[0258] For example, the implant can be implanted in the extrascleral or intrascleral space, on the sclera, in the vitreous cavity, or in the anterior chamber. For example, subconjunctival, intrascleral, and anterior chamber implants can be used to treat anterior segment diseases, and for example, intravitreal, suprachoroidal, and intrascleral implants can be used to treat posterior segment diseases.

[0259] In some embodiments, formulations of lipid binding protein-based complexes, such as CER-001 and optionally one or more ophthalmic drugs, are formulated for delivery via iontophoresis, such as transcorneal or transscleral iontophoresis.

[0260] In other embodiments, formulations of lipid binding protein-based complexes, such as CER-001 and optionally one or more ophthalmic drugs, are formulated for delivery via electroporation.

[0261] CER-001 intended for administration by infusion can be formulated in a phosphate buffer containing sucrose and mannitol excipients, for example as described in WO 2012 / 109162. Formulations of lipid-binding protein-based complexes for topical administration may include, for example, carriers, stabilizers, excipients, and combinations thereof. Topical formulations (e.g., eye drops) may include buffers such as phosphate, citrate, or other inorganic acid buffers, antioxidants such as ascorbic acid and / or methionine, preservatives, low molecular weight polypeptides, proteins such as gelatin, serum albumin, or immunoglobulins, hydrophilic polymers such as PVP, amino acids, monosaccharides or disaccharides or other carbohydrates, chelating agents, sugars, nonionic surfactants, and the like.

[0262] In some embodiments, the topical formulation comprises an osmotic pressure regulator. In some embodiments, the osmotic pressure regulator is sodium chloride.

[0263] In some embodiments, the topical formulation comprises a preservative. In some embodiments, the preservative is benzalkonium chloride, cetrimonium, sodium perborate, stabilized oxychlorine complex, SofZia, polyquaternium-1, chlorobutanol, disodium edetate, polyhexamethylene biguanide, or a combination thereof.

[0264] In some embodiments, the topical formulation comprises a buffer. In some embodiments, the buffer is selected from borates, borate-polyol complexes, succinates, phosphate buffers, citrate buffers, acetate buffers, carbonate buffers, organic buffers, amino acid buffers, and combinations thereof.

[0265] In some embodiments, the topical formulation comprises a tonicity modifier. In some embodiments, the tonicity modifier is selected from sodium chloride, sodium nitrate, sodium sulfate, sodium bisulfate, potassium chloride, calcium chloride, magnesium chloride, zinc chloride, potassium acetate, sodium acetate, sodium bicarbonate, sodium carbonate, sodium thiosulfate, magnesium sulfate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dextrose, mannitol, sorbitol, dextrose, sucrose, urea, propylene glycol, glycerol, trehalose, and combinations thereof.

[0266] The formulation of lipid-binding protein-based complexes (e.g., CER-001) intended for intraocular administration may include, for example, carriers, stabilizers, viscosity enhancers, osmotic pressure regulators, buffers, and combinations thereof. In some embodiments, the intraocular formulation comprises an osmotic pressure regulator. An example of an osmotic pressure regulator is sodium chloride. In some embodiments, the intraocular formulation comprises a buffer. Examples of buffers include borates, borate-polyol complexes, succinates, phosphate buffers, citrate buffers, acetate buffers, carbonate buffers, organic buffers, amino acid buffers, and combinations thereof.

[0267] In some embodiments, a formulation comprising CER-001 (optionally, wherein CER-001 is used as a carrier for one or more ophthalmic drugs) may comprise CER-001 at a concentration of 0.5 mg / ml to 8 mg / ml (e.g., 0.5 mg / ml, 0.8 mg / ml, 1 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, any range bounded by any two of the foregoing values) based on protein weight. In some embodiments, a formulation comprising CER-001 may comprise CER-001 at a concentration of at least 1 mg / ml, at least 2 mg / ml, at least 3 mg / ml, at least 4 mg / ml, at least 5 mg / ml, at least 6 mg / ml, at least 7 mg / ml, or at least 8 mg / ml (based on protein weight).

[0268] The formulation of a lipid-binding protein-based complex (e.g., CER-001) and one or more ophthalmic drugs can be produced, for example, by thermal cycling a mixture containing a lipid-binding protein-based complex and one or more ophthalmic drugs. For example, the mixture can be (a) heated from a temperature within a first temperature range to a temperature within a second temperature range, then (b) cooled from a temperature within the second temperature range to a temperature within the first temperature range, and then (c) optionally subjected to one or more additional heating and cooling cycles, for example, a total of two, three, four, five or six heating and cooling cycles. Alternatively, the mixture (a) can be cooled from a temperature within the second temperature range to a temperature within the first temperature range, then (b) heated from a temperature within the first temperature range to a temperature within the second temperature range, and then (c) optionally subjected to one or more additional cooling and heating cycles, for example, a total of two, three, four, five or six cooling and heating cycles. In some embodiments, the first temperature range can include from 30°C to 45°C (e.g., 35°C to 45°C, 30°C to 35°C, 35°C to 40°C or 40°C to 45°C). In some embodiments, the second temperature range may include a temperature from 50°C to 65°C (e.g., 50°C to 60°C, 50°C to 55°C, or 55°C to 60°C). In some embodiments, the thermal cycle includes thermal cycling between 37°C and 55°C, for example as described in Example 1. Thus, in some aspects, the present disclosure provides a composition comprising a lipid-binding protein-based complex (e.g., CER-001) and one or more ophthalmic drugs by a method comprising thermal cycling a mixture comprising the lipid-binding protein-based complex and one or more ophthalmic drugs.

[0269] 6.2. Subject population

[0270] The subject that can be treated according to the methods described herein is preferably a mammal, most preferably a human.

[0271] In some respects, the subject can be a subject who needs to treat an eye disease, such as an eye disease relevant to lipid accumulation, such as an eye lipid deposition. In some cases, lipid may be accumulated in eyes or near eyes. The exemplary eye disease relevant to lipid accumulation that can be treated by the disclosed method comprises cataract, dry eye disease, such as dry eye disease relevant to meibomian gland dysfunction or tear gland dysfunction, blepharitis, uveitis, corneal disease such as lipid keratopathy (for example, secondary lipid keratopathy, such as lipid keratopathy secondary to previous eye disease or damage) and corneal dystrophy (for example, hereditary corneal dystrophy, anterior or superficial corneal dystrophy, stromal corneal dystrophy or posterior corneal dystrophy), the eye disease relevant to LCAT defect, such as fish eye disease, dry macular degeneration (dry AMD), Stargardt disease and Leber idiopathic stellate neuroretinitis. For example, in a subject suffering from LCAT defect, lipid deposition in cornea can cause damage, such as blurred vision.

[0272] In some aspects, the subject has cataracts. For clarity, treatment of a subject having cataracts includes treatment of a subject having unilateral cataracts and a subject having bilateral cataracts.

[0273] In some aspects, the subject suffers from an inflammatory eye disease, such as uveitis (e.g., anterior uveitis, intermediate uveitis, posterior uveitis, or panuveitis) or scleritis. In some aspects, the subject suffers from acute anterior uveitis.

[0274] In some embodiments, the subject treated according to the method and / or dosage regimen of the present disclosure suffers from LCAT deficiency, and optionally the lipid-binding protein-based complex for treating the subject is CER-001. The subject can be a homozygote or heterozygote of LCAT mutation. In some embodiments, the subject treated according to the method and / or dosage regimen of the present invention suffers from fish eye disease. Subjects suffering from fish eye disease will usually have bilateral corneal opacities, and may have visual impairment, for example, reduced contrast sensitivity and / or blurred vision compared to normal. Corneal opacity and its progression or disappearance (for example, in response to treatment as described herein) can be qualitatively assessed, for example, by comparing the slit lamp images taken at different times by the subject's eyes. Corneal opacity can also be quantitatively assessed, for example, by anterior segment optical coherence tomography (OCT) (see Kanai et al., 2018, American Journal of Ophthalmology Case Reports, 10: 137-141, incorporated herein by reference in its entirety). For example, visual function can be assessed by measuring the contrast sensitivity of the subject using a standard test chart (e.g., CSV-1000E chart; Vector Vision Co., Greenville, OH). Stray light measurement can be used to quantify light scattering that creates a stray light overlay on the retinal image, which can lead to increased blurred vision or glare disturbances. Stray light can be measured using a stray light meter (e.g., C-Quant from Oculus GmbH, Wetzlar, Germany). In certain embodiments, the methods of the present disclosure can reduce the severity of fish eye disease in a subject, for example, as measured by corneal opacity, contrast sensitivity, stray light value, or a combination thereof.

[0275] In some embodiments, the subject does not have LCAT deficiency.

[0276] In some embodiments, the subject suffers from an eye disease other than fish eye disease, such as an eye disease other than fish eye disease described herein, and optionally, wherein the lipid binding protein-based complex used to treat the subject is CER-001.

[0277] In some embodiments, the subject suffers from a genetic disease such as Stargardt disease, optionally wherein the lipid binding protein-based complex used to treat the subject is CER-001.

[0278] In some embodiments, the subject suffers from macular degeneration, such as dry AMD or wet AMD, and optionally, the lipid binding protein-based complex used to treat the subject is CER-001. In other embodiments, the subject suffers from an eye disease other than macular degeneration, such as an eye disease other than macular degeneration described herein.

[0279] In some embodiments, the subject has diabetic retinopathy, optionally wherein the lipid binding protein-based complex used to treat the subject is CER-001. In some embodiments, the subject having diabetic retinopathy has diabetic macular edema.

[0280] In some embodiments, the subject has retinal vein occlusion.

[0281] In some embodiments, the subject suffers from dry eye (e.g., severe dry eye). In some embodiments, the subject suffers from meibomian gland dysfunction (MBD), such as obstructive MGD. In other embodiments, the subject suffers from lacrimal gland dysfunction. In some embodiments, the subject suffers from blepharitis. In some embodiments, the subject suffers from uveitis (e.g., caused by bacterial infection). In some embodiments, the subject suffers from lipid keratopathy. In some embodiments, the lipid-binding protein-based complex for treating a subject suffering from one of the eye diseases described in this paragraph is CER-001.

[0282] In some embodiments, the subject has ocular lipid deposition, which includes lipid deposition present in and / or near the eye. In one embodiment, the lipid deposition is corneal lipid deposition, retinal lipid deposition, eyelid lipid deposition, or a combination thereof. In some embodiments, the lipid-binding protein-based complex used to treat a subject suffering from one of the eye diseases described in this paragraph is CER-001.

[0283] In some embodiments, the subject has lipid deposits in the cornea and / or retina. Lipid deposits in the cornea can lead to visual impairment, such as blurred vision. Lipid deposits in the retina, such as drusen in dry AMD or lipofuscin in Stargardt's disease, can lead to retinal degeneration. In some embodiments, the lipid-binding protein-based complex used to treat a subject suffering from one of the eye diseases described in this paragraph is CER-001.

[0284] In some embodiments, the subject has eyelid lipidosis, which is lipid deposition on the eyelids.

[0285] In some embodiments, lipid deposition is in drusen deposition.Drusen is the focal (focal) deposition of extracellular debris between the basal layer of retinal pigment epithelium (RPE) and the collagen layer in Bruch's membrane.Most drusen are hard, dome-shaped, solid inside, homogeneous contents, and a median diameter of 47 μm.Hard drusen include lipid particles of about 60-90nm in diameter, which contain abundant esterified cholesterol, unesterified cholesterol, phosphatidylcholine and apolipoprotein B.With the increase of age, it is normal for some hard drusen to occur.The presence of larger and more drusen in the macula is a common early sign of age-related macular degeneration (AMD).

[0286] In one embodiment, lipid deposition is lipofuscin granules. Lipofuscin granules accumulate in the RPE lysosomal compartment after mitosis. Lipofuscin granules mainly contain N-retinylidene-N-retinyl-ethanolamine (A2E). The presence of lipofuscin granules is a sign of Stargardt disease.

[0287] In one embodiment, lipid deposition is cholesterol deposition, especially cholesterol deposition in cornea. In the individuality with LCAT genetic defect, cholesterol accumulates in the extracellular connective tissue matrix of corneal stroma. Usually, the diameter of this type of cholesterol deposition is 0.2 to 2.5 μm.

[0288] In one embodiment, the ocular lipid deposits are not calcified.

[0289] The presence of ocular lipid deposits can be determined by one or more of retinal slit lamp photography, Heidelberg retinal tomography (HRT) scanning, optical coherence tomography (OCT), fundus autofluorescence imaging, slit lamp fundus. In particular, drusen can be observed by retinal slit lamp photography, HRT scanning and / or OCT; lipofuscin particles can be observed by fundus autofluorescence imaging; and cholesterol deposits in the cornea can be observed by slit lamp.

[0290] Use of the lipid binding protein complexes described herein can reduce the severity of eye disease in a subject. Without being bound by theory, it is believed that the lipid binding protein complexes can solubilize lipids accumulated in ocular deposits, leading to their elimination.

[0291] In some embodiments, the use of the lipid binding protein complexes described herein can reduce the number of ocular lipid deposits. In some embodiments, the use of the lipid binding protein complexes described herein can reduce the size of ocular lipid deposits.

[0292] The reduction in the number and / or size of lipid deposits can be qualitatively assessed, for example, by comparing the results of examinations performed before and during or after administration of the lipid-binding protein complex, such as slit lamp photography of the retina, Heidelberg retinal tomography (HRT) scans, optical coherence tomography (OCT), fundus autofluorescence imaging, slit lamp fundus. The reduction in the number and / or size of lipid deposits can also be quantitatively assessed by the above methods.

[0293] Alternatively, a reduction in the number and / or size of lipid deposits can be determined indirectly by comparing measurements of corneal opacity, contrast sensitivity, stray light values, or a combination thereof, obtained before and during or after administration of the lipoprotein complexes.

[0294] In certain embodiments, a reduction in severity of an eye disease can be measured, for example, by assessing corneal opacity, contrast sensitivity, stray light values, or a combination thereof.

[0295] In one embodiment, the subject has impaired vision, including blurred vision, due to ocular lipid deposition, and the amount of the lipoprotein complex is an amount that improves the subject's vision.

[0296] In one embodiment, the subject suffers from corneal opacity due to lipid deposition in the cornea. Treatment with lipid-binding protein complexes as described herein can reduce the opacity of the subject's cornea. Corneal opacity and its progression or regression (e.g., in response to treatment as described herein) can be qualitatively assessed, such as by comparing slit lamp images taken at different times by the subject's eyes. Corneal opacity can also be quantitatively assessed, such as by anterior segment optical coherence tomography (OCT) (see Kanai et al., 2018, American Journal of Ophthalmology Case Reports, 10: 137-141, incorporated herein by reference in its entirety). For example, visual function can be assessed by measuring the contrast sensitivity of the patient using a standard test chart (e.g., CSV-1000E chart; VectorVision Co., Greenville, OH). Stray light measurement can be used to quantify the light scattering that produces a stray light cover on the retinal image, which can cause blurred vision or increased glare disorders. Stray light can be measured using a stray light meter (e.g., C-Quant from Oculus GmbH, Wetzlar, Germany).

[0297] In one embodiment, the amount of lipid binding protein complex administered is an amount effective to reduce corneal opacity in a patient.

[0298] In one embodiment, the amount of lipid binding protein complex administered is effective to improve contrast sensitivity in the patient.

[0299] In one embodiment, the amount of lipid binding protein complex administered is effective to reduce the stray light value in the patient.

[0300] In other aspects, the subject suffers from an eye disease (which may be, but not necessarily, a disease associated with lipid accumulation) and the lipid-binding protein-based complex is used as a drug carrier to deliver one or more ophthalmic drugs to the subject's eye. For example, the subject may have an anterior or posterior eye condition, such as uveitis (e.g., caused by bacterial infection), macular edema (e.g., diabetic macular edema), macular degeneration, retinal detachment, an eye tumor, a fungal infection, a viral infection, a bacterial infection, such as bacterial conjunctivitis or trachoma, multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis, or glaucoma.

[0301] 6.2.1. CER-001 for the treatment of uveitis

[0302] Inflammatory eye diseases, such as uveitis (e.g., anterior uveitis, intermediate uveitis, posterior uveitis, or panuveitis), which may or may not be caused by bacterial infection, can be treated by administering CER-001. The use of CER-001 to treat uveitis can be accomplished by administering a therapeutically effective amount of CER-001 to a subject in need thereof, such as an amount that reduces the severity of uveitis (e.g., by alleviating one or more symptoms of uveitis). CER-001 can be appropriately formulated according to the intended route of administration. Exemplary formulations are described in Section 6.1.10. For the treatment of uveitis, topical administration of CER-001, such as topical administration or intraocular administration to a subject in need thereof, is preferred. Intraocular administration can be, for example, intraocular injection, such as intravitreal injection, subconjunctival injection, juxtabulbar injection, peribulbar injection, retrobulbar injection, suprachoroidal injection, or suprascleral injection. Implants can also be used to deliver CER-001. For topical administration, CER-001 can be administered, for example, as eye drops. As shown in Examples 3 and 4, good ocular tolerability of CER-001 was observed even with repeated administrations.

[0303] In some embodiments, CER-001 may be used to treat uveitis in a subject in need thereof according to a dosing regimen described in one or more of Sections 6.3, 6.4, and 6.5. For example, CER-001 may be used to treat uveitis in a subject in need thereof according to an induction regimen described in Section 6.3. Alternatively, or in addition, CER-001 may be used to treat uveitis in a subject in need thereof according to a consolidation regimen described in Section 6.4. Alternatively, or in addition, CER-001 may be used to treat uveitis in a subject in need thereof according to a maintenance regimen described in Section 6.5. In some embodiments, CER-001 may be used to treat uveitis in a subject in need thereof according to an induction regimen described in Section 6.3; and / or a consolidation regimen described in Section 6.4; and / or a maintenance regimen described in Section 6.5.

[0304] 6.2.2. CER-001 and ophthalmic drugs for the treatment of eye diseases

[0305] In some embodiments, inflammatory eye diseases such as uveitis (e.g., anterior uveitis (e.g., acute anterior uveitis), intermediate uveitis, posterior uveitis, or panuveitis), which may or may not be caused by bacterial infection, can be treated by administering a composition comprising CER-001 and dexamethasone, a composition comprising CER-001 and dexamethasone palmitate, or a composition comprising CER-001 and tacrolimus. The use of such compositions for treating uveitis can be accomplished by administering a therapeutically effective amount of the composition to a subject in need thereof, e.g., in an amount that reduces the severity of uveitis (e.g., by alleviating one or more symptoms of uveitis). The composition can be appropriately formulated according to the intended route of administration. Exemplary formulations are described in Section 6.1.10. For the treatment of uveitis, topical administration of the composition, e.g., topical administration or intraocular administration to a subject in need thereof is preferred. Intraocular administration can be, for example, intraocular injection, such as intravitreal injection, subconjunctival injection, juxtabulbar injection, peribulbar injection, retrobulbar injection, suprachoroidal injection or suprascleral injection. Implants can also be used. For topical administration, the composition can be administered as, for example, eye drops. As shown in Examples 3 and 4, good ocular tolerance of such compositions was observed even with repeated administration.

[0306] In some embodiments, a composition comprising CER-001 and dexamethasone, a composition comprising CER-001 and dexamethasone palmitate, or a composition comprising CER-001 and tacrolimus can be used to treat uveitis in a subject in need thereof according to a dosing regimen described in one or more of Sections 6.3, 6.4, and 6.5. For example, the composition can be used to treat uveitis in a subject in need thereof according to an induction regimen described in Section 6.3. Alternatively, or in addition, the composition can be used to treat uveitis in a subject in need thereof according to a consolidation regimen described in Section 6.4. Alternatively, or in addition, the composition can be used to treat uveitis in a subject in need thereof according to a maintenance regimen described in Section 6.5. In some embodiments, the composition can be used to treat uveitis in a subject in need thereof according to an induction regimen described in Section 6.3; and / or a consolidation regimen described in Section 6.4; and / or a maintenance regimen described in Section 6.5.

[0307] Compositions comprising CER-001 and dexamethasone and compositions comprising CER-001 and dexamethasone palmitate may also be used to treat other eye diseases, such as macular degeneration and dry macular edema (e.g., by alleviating one or more symptoms of the disease). Compositions comprising CER-001 and tacrolimus may also be used to treat other eye diseases, such as dry eye disease, such as severe dry eye disease (e.g., by alleviating one or more symptoms of the disease).

[0308] Compositions comprising CER-001 and dexamethasone, compositions comprising CER-001 and dexamethasone palmitate, and compositions comprising CER-001 and tacrolimus can be prepared in some embodiments by thermal cycling a mixture comprising CER-001 and the corresponding drug, for example as described in Section 6.1.10.

[0309] 6.3. Induction regimen

[0310] Induction regimens suitable for use in the methods of the present disclosure require administration of multiple doses of the lipid binding protein-based complex (eg, CER-001) with one or more days between each administration.

[0311] The induction regimen typically includes at least three doses of the lipid binding protein-based complex (e.g., CER-001), but may include four or more doses of the lipid binding protein-based complex (e.g., CER-001), for example, five, six, seven, eight, nine, ten, eleven or twelve doses.

[0312] The induction regimen may last for one or more weeks, two or more weeks, three or more weeks, four or more weeks, five or more weeks, six or more weeks, seven or more weeks, eight or more weeks, nine or more weeks, or ten or more weeks.

[0313] For example, an induction regimen may include administration of:

[0314] three doses of a lipid-binding protein-based complex (e.g., CER-001) in one week;

[0315] three doses of lipid-binding protein-based complexes (e.g., CER-001) over two weeks;

[0316] three doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0317] four doses of a lipid-binding protein-based complex (e.g., CER-001) over two weeks;

[0318] four doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0319] five doses of a lipid-binding protein-based complex (e.g., CER-001) over two weeks;

[0320] five doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0321] five doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0322] six doses of a lipid-binding protein-based complex (e.g., CER-001) over two weeks;

[0323] six doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0324] six doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0325] seven doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0326] seven doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0327] seven doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0328] eight doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0329] eight doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0330] eight doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0331] nine doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0332] nine doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0333] nine doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0334] nine doses of a lipid-binding protein-based complex (e.g., CER-001) over six weeks;

[0335] Ten doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0336] Ten doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0337] Ten doses of a lipid binding protein-based complex (e.g., CER-001) over six weeks; or

[0338] Ten doses of a lipid binding protein-based complex (eg, CER-001) over seven weeks.

[0339] In embodiments, the induction regimen comprises two doses per week of the lipid binding protein-based complex (eg, CER-001) to five doses per week.

[0340] In embodiments, the induction regimen comprises administering 9 doses of the lipid binding protein-based complex (eg, CER-001) over three weeks, eg, on days 1, 2, 4, 7, 9, 11, 14, 16, and 18.

[0341] In practice, an administration window can be provided, for example, to accommodate slight changes in a dosing regimen of multiple doses per week. For example, a window of ±2 days or ±1 day before and after the dosing date can be used.

[0342] The therapeutic dose range of the lipid-binding protein-based complex (e.g., CER-001) administered by infusion in an induction regimen can be 4 to 30 mg / kg based on protein weight (e.g., 4, 5, 6, 7, 8, 9, 10, 12, 15, 20, 25 or 30 mg / kg, or a range defined by any two of the above values, such as 5 to 15 mg / kg, 10 to 20 mg / kg or 15 to 25 mg / kg). As used herein, the expression "based on protein weight" refers to the dose of the lipid-binding protein-based complex (e.g., CER-001) to be administered to a subject, which is calculated based on the amount of lipid-binding protein (e.g., ApoA-I) in the lipid-binding protein-based complex (e.g., CER-001) to be administered and the body weight of the subject. For example, a subject weighing 70 kg and receiving a 10 mg / kg dose of CER-001 will receive CER-001 in an amount providing 700 mg ApoA-I (70 kg x 10 mg / kg). In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the induction regimen is 8 mg / kg. In some embodiments, the induction regimen includes nine doses of lipid-binding protein-based complexes (e.g., CER-001) administered at a dose of 8 mg / kg over three weeks. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the induction regimen is 10 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the induction regimen is 15 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the induction regimen is 20 mg / kg. In some embodiments, the induction regimen includes nine doses of the lipid binding protein-based complex (eg, CER-001) administered at a dose of 10 mg / kg over three weeks. The dose of the lipid binding protein-based complex for delivering an ophthalmic drug may be a dose that delivers a therapeutically effective amount of the drug.

[0343] In other aspects, the lipid binding protein-based complex (eg, CER-001) can be administered on a unit dose basis. The unit dose used during the induction phase can vary from 300 mg to 3000 mg per administration by infusion.

[0344] In specific embodiments, the dose of the lipid binding protein-based complex (eg, CER-001) used during the induction phase is 300 mg to 1500 mg, 400 mg to 1500 mg, 500 mg to 1200 mg, or 500 mg to 1000 mg per administration by infusion.

[0345] In a particular embodiment, the lipid-binding protein-based complex (e.g., CER-001) is administered as an IV infusion. For example, a stock solution of CER-001 can be diluted in normal saline (e.g., normal saline (0.9% NaCl)) to a total volume of between 125 and 250 ml. In a preferred embodiment, a subject weighing less than 80 kg will have a total volume of 125 ml, while a subject weighing at least 80 kg will have a total volume of 250 ml. The lipid-binding protein-based complex (e.g., CER-001) can be administered over one hour using an infusion pump at a fixed rate of 250 ml / hr. Depending on the needs of the subject, it can be administered by a slow infusion lasting more than one hour (e.g., up to two hours), by a rapid infusion of one hour or less, or by a single bolus.

[0346] In an optional embodiment, the lipid-binding protein-based complex (e.g., CER-001) is topically administered to the eye, for example, by intraocular injection or topical administration. A storage solution of the lipid-binding protein-based complex (e.g., CER-001) can be diluted in a suitable diluent before administration. Suitable diluents include normal saline, such as physiological saline (0.9% NaCl). In some embodiments, the lipid-binding protein-based complex is formulated as eye drops.

[0347] 6.4. Consolidation plan

[0348] Consolidation regimens suitable for use in the methods of the present disclosure entail administration of multiple doses of the lipid binding protein-based complex (eg, CER-001) with 1 day or more between each dose, eg, 2 days or more between each administration.

[0349] The consolidation regimen typically includes at least two doses of the lipid-binding protein-based complex (e.g., CER-001), but may include three or more doses of the lipid-binding protein-based complex (e.g., CER-001), for example, four, five, six, seven, eight, nine or ten.

[0350] The consolidation regimen may last for one or more weeks, two or more weeks, three or more weeks, four or more weeks, five or more weeks, six or more weeks, seven or more weeks, eight or more weeks, nine or more weeks, or ten or more weeks.

[0351] For example, a consolidation regimen may include administration of:

[0352] Two doses of a lipid-binding protein-based complex (e.g., CER-001) within one week;

[0353] Two doses of a lipid-binding protein-based complex (e.g., CER-001) within two weeks;

[0354] three doses of lipid-binding protein-based complexes (e.g., CER-001) over two weeks;

[0355] three doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0356] four doses of a lipid-binding protein-based complex (e.g., CER-001) over two weeks;

[0357] four doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0358] five doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0359] five doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0360] five doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0361] six doses of a lipid-binding protein-based complex (e.g., CER-001) over three weeks;

[0362] six doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0363] six doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0364] seven doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0365] seven doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0366] seven doses of a lipid-binding protein-based complex (e.g., CER-001) over six weeks;

[0367] eight doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0368] eight doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0369] eight doses of a lipid-binding protein-based complex (e.g., CER-001) over six weeks;

[0370] nine doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0371] nine doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0372] nine doses of a lipid-binding protein-based complex (e.g., CER-001) over six weeks;

[0373] nine doses of a lipid-binding protein-based complex (e.g., CER-001) over six weeks;

[0374] Ten doses of a lipid-binding protein-based complex (e.g., CER-001) over four weeks;

[0375] Ten doses of a lipid-binding protein-based complex (e.g., CER-001) over five weeks;

[0376] Ten doses of a lipid binding protein-based complex (e.g., CER-001) over six weeks; or

[0377] Ten doses of a lipid binding protein-based complex (eg, CER-001) over seven weeks.

[0378] In embodiments, the consolidation regimen includes two doses of the lipid binding protein-based complex (eg, CER-001) per week to five doses per week.

[0379] In embodiments, the consolidation regimen comprises administering six doses of the lipid binding protein-based complex (eg, CER-001) over three weeks, eg, on days 21, 24, 28, 31, 35, and 38 of an induction regimen beginning on day 1.

[0380] In practice, an administration window can be provided, for example, to accommodate slight changes in a dosing regimen of multiple doses per week. For example, a window of ±2 days or ±1 day before and after the dosing date can be used.

[0381] The therapeutic dose range of the lipid-binding protein-based complex (e.g., CER-001) administered by infusion in the consolidation regimen can be 4 to 30 mg / kg based on the weight of the protein (e.g., 4, 5, 6, 7, 8, 9, 10, 12, 15, 20, 25 or 30 mg / kg, or a range defined by any two of the above values, such as 5 to 15 mg / kg, 10 to 20 mg / kg, or 15 to 25 mg / kg). In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the consolidation regimen is 8 mg / kg. In some embodiments, the consolidation regimen includes administering six doses of the lipid-binding protein-based complex (e.g., CER-001) at a dose of 8 mg / kg over three weeks. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the consolidation regimen is 10 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) in the consolidation regimen is 15 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the consolidation regimen is 20 mg / kg. In some embodiments, the consolidation regimen includes administering six doses of the lipid-binding protein-based complex (e.g., CER-001) at a dose of 10 mg / kg over three weeks. The dose of the lipid-binding protein-based complex for delivering an ophthalmic drug can be a dose that delivers a therapeutically effective amount of the drug.

[0382] In other aspects, the lipid binding protein-based complex (eg, CER-001) can be administered on a unit dose basis. The unit dose for the consolidation phase can vary from 300 mg to 3000 mg per administration by infusion.

[0383] In specific embodiments, the dose of the lipid binding protein-based complex (eg, CER-001) used during the consolidation phase is 300 mg to 1500 mg, 400 mg to 1500 mg, 500 mg to 1200 mg, or 500 mg to 1000 mg per administration by infusion.

[0384] In some embodiments, the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered during the consolidation phase is greater than the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered during the induction phase. For example, the dosage administered in the consolidation phase can be 1.5 to 3 times the dosage administered in the induction phase. In a specific embodiment, the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered during the consolidation phase is 2 times the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered during the induction phase. Increasing the dose during the consolidation phase can offset the reduction in dosing frequency. In other embodiments, the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered during the consolidation phase is the same as the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered during the induction phase.

[0385] The lipid-binding protein-based complex (e.g., CER-001) can be administered in the same manner as described in Section 6.3 during the consolidation phase, for example, as an IV infusion or topically administered, such as intraocular or topical administration, within one hour. When the dose of the lipid-binding protein-based complex (e.g., CER-001) administered during the consolidation phase is greater than the dose administered during the induction phase, the lipid-binding protein-based complex (e.g., CER-001) can optionally be administered and / or infused in a larger volume and / or administered for a longer period of time. For example, when the dose of the lipid-binding protein-based complex (e.g., CER-001) administered during the consolidation phase is twice the dose administered during the induction phase, the volume of administration can be increased (e.g., doubled) and / or the infusion time can be increased (e.g., doubled).

[0386] 6.5. Maintenance plan

[0387] The methods of the present disclosure may include a maintenance regimen, which may, but not necessarily, be after an induction regimen and an optional consolidation regimen. In some embodiments, the maintenance regimen includes administering a lipid-binding protein-based complex (e.g., CER-001) to a subject at a frequency lower than during the induction phase and / or consolidation phase. Typically, a lipid-binding protein-based complex (e.g., CER-001) is administered once every 3 or more days during the maintenance regimen, such as once a week or twice a week.

[0388] A maintenance regimen may entail administering a lipid binding protein-based complex (e.g., CER-001) for one month or more, two months or more, three months or more, six months or more, nine months or more, one year or more, 18 months or more, two years or more, or indefinitely.

[0389] In some embodiments, the maintenance regimen includes administering a lipid-binding protein-based complex (e.g., CER-001) once every 5 days to a week for at least 16 weeks. In other embodiments, the maintenance regimen includes administering a lipid-binding protein-based complex (e.g., CER-001) once a week for at least 20 weeks, at least 30 weeks, or at least 40 weeks.

[0390] Similar to the administration windows described in Section 6.3 above, administration windows can also be used to maintain a regimen to accommodate slight changes in the weekly dosing regimen. For example, a window of ±2 days or ±1 day before and after a weekly date can be used.

[0391] The therapeutic dose range of the lipid-binding protein-based complex (e.g., CER-001) administered by infusion in the maintenance regimen is 4 to 30 mg / kg based on the weight of the protein (e.g., 4, 5, 6, 7, 8, 9, 10, 12, 15, 20, 25 or 30 mg / kg, or any range bounded by any two of the above values, such as 5 to 15 mg / kg, 10 to 20 mg / kg or 15 to 25 mg / kg). For example, a subject weighing 70 kg and receiving a 10 mg / kg dose of CER-001 will receive CER-001 in an amount providing 700 mg ApoA-I (70 kg x 10 mg / kg). In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the maintenance regimen is 8 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the maintenance regimen is 10 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the maintenance regimen is 15 mg / kg. In some embodiments, the dose of the lipid-binding protein-based complex (e.g., CER-001) used in the maintenance regimen is 20 mg / kg. The dose of the lipid-binding protein-based complex for delivering an ophthalmic drug can be a dose that delivers a therapeutically effective amount of the drug.

[0392] In other aspects, the lipid binding protein-based complex (eg, CER-001) can be administered on a unit dose basis. The unit dose for the maintenance phase can vary from 300 mg to 3000 mg per administration by infusion.

[0393] In specific embodiments, the dose of the lipid binding protein-based complex (eg, CER-001) used during the maintenance phase is 300 mg to 1500 mg, 400 mg to 1500 mg, 500 mg to 1200 mg, or 500 mg to 1000 mg per administration by infusion.

[0394] In some embodiments, the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered in the maintenance phase is greater than the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered during the induction phase and / or the consolidation phase. For example, the dosage administered in the maintenance phase can be 1.5 to 3 times the dosage administered in the consolidation phase. In a specific embodiment, the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered in the maintenance phase is 2 times the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered in the consolidation phase. Increasing the dose in the maintenance phase can offset the reduction in the frequency of administration. In other embodiments, the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered in the maintenance phase is the same as the dosage of the lipid-binding protein-based complex (e.g., CER-001) administered in the induction phase and / or the consolidation phase. In some embodiments, the dosage administered in the maintenance phase can be adjusted, such as increased or decreased, such as to achieve a dose of a stable clinical parameter (e.g., corneal opacity). Alternatively or additionally, the frequency of administration during the maintenance phase can be adjusted, eg, increased or decreased, eg, to achieve stabilization of a clinical parameter (eg, corneal opacity).

[0395] The lipid-binding protein-based complex (e.g., CER-001) can be administered in the same manner as described in Section 6.3 during the maintenance phase, for example, as an IV infusion or topical administration, such as intraocular or topical administration. When the dose of the lipid-binding protein-based complex (e.g., CER-001) administered during the maintenance phase is greater than the dose administered during the consolidation phase, the lipid-binding protein-based complex (e.g., CER-001) can optionally be administered and / or infused in a larger volume and / or administered over a longer period of time. For example, when the dose of the lipid-binding protein-based complex (e.g., CER-001) administered during the maintenance phase is twice the dose administered during the consolidation phase, the administration volume can be increased (e.g., doubled) and / or the infusion time can be increased (e.g., doubled).

[0396] 6.6. Combination therapy

[0397] A subject can be treated with a lipid binding protein-based complex (e.g., CER-001) as a monotherapy or as part of a combination therapy regimen, for example, with one or more lipid-control drugs such as statins (e.g., atorvastatin, rosuvastatin, simvastatin, fluvastatin, lovastatin, pravastatin), cholesterol absorption inhibitors (e.g., ezetimibe), niacin, aspirin, proprotein convertase subtilisin / kexin type 9 (PCSK9) inhibitors (e.g., antibodies such as alirocumab, bococizumab, evolocumab, 1D05-IgG2 (NNi et al., 2011, J Lipid Res. 52(1):78-86) and LY3015014 (Kastelein et al., 2016, Eur Heart J 37(17):1360-9) or RNAi therapeutics such as ALN-PCSSC (Medicines Company) or antihypertensive drugs (e.g., amlodipine, urapidil, furosemide, and combinations thereof). For example, a subject with fish eye disease can be treated with a combination of one or more lipid-controlling drugs and a lipid-binding protein-based complex. In some embodiments, the combination therapy includes a lipid-binding protein-based complex (e.g., CER-001) in combination with a standard of care treatment for the subject's eye disease.

[0398] Combination therapy regimens may require administration of lipid-binding protein-based complexes (e.g., CER-001) and one or more of the aforementioned drugs and / or one or more of the aforementioned drug classes. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with atorvastatin. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with ezetimibe. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with niacin. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with rosuvastatin. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with simvastatin. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with aspirin. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with fluvastatin. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with lovastatin. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with pravastatin. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with alikazumab. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with ivakuzumab. In some embodiments, a subject is treated with a lipid-binding protein-based complex (e.g., CER-001) in combination with ALN-PCSsc. In each of the aforementioned embodiments, the lipid-control drug can be the only lipid-control drug combined with a lipid-binding protein-based complex therapy that the subject receives, or can be part of a combination of lipid-control drugs administered with a lipid-binding protein-based complex.

[0399] In some embodiments, the lipid-binding protein-based complex (e.g., CER-001) is administered in combination with an antihypertensive drug, for example, one, two, or all three of amlodipine, urapidil, and furosemide. In some embodiments, the lipid-binding protein-based complex (e.g., CER-001) is administered in combination with amlodipine, urapidil, and furosemide. In some embodiments, the combination further comprises a statin, such as atorvastatin.

[0400] The lipid-binding protein-based complex (eg, CER-001)-based therapy may be added to background lipid-lowering therapy prior to treatment with the lipid-binding protein-based complex (eg, CER-001).

[0401] In some embodiments, the subject is treated with a stable dose of a lipid-controlling drug for at least 6 weeks (e.g., 6 weeks, 8 weeks, 2 months, 6 months, 1 year, or more than 1 year) prior to starting therapy with a lipid-binding protein-based complex (e.g., CER-001) according to a dosing regimen disclosed herein. Alternatively, therapy with a lipid-binding protein-based complex (e.g., CER-001) may be started prior to or concurrently with treatment with one or more lipid-controlling drugs. 7. Examples

[0402] 7.1. Example 1: CER-001 as a carrier for ophthalmic drugs

[0403] 7.1.1. Azithromycin and spironolactone

[0404] Azithromycin is an antibiotic used to treat bacterial infections of the eye, such as bacterial conjunctivitis and trachoma (www.mayoclinic.org / drugs-supplements / azithromycin-ophthalmic-route / description / drg-20070979). Spironolactone is a steroid that has been studied for the treatment of meibomian gland dysfunction and associated dry eye (Yee et al., 2016, Investigative Ophthalmology & Visual Science 57(12):5664). A study was conducted to evaluate the suitability of CER-001 as a drug carrier to deliver azithromycin and spironolactone.

[0405] The CER-001 solution was mixed with azithromycin to provide a final azithromycin concentration of 10 mg / ml and subjected to five heating and cooling cycles between 37°C and 55°C to promote the complexation of azithromycin with CER-001. For control, a CER-001 sample without azithromycin added and a sample of azithromycin at a concentration of 10 mg / ml in phosphate buffered saline (PBS) were also subjected to five heating and cooling cycles between 37°C and 55°C. Figure 1A As shown, the CER-001 sample without azithromycin (left tube) remained clear after the heating and cooling cycles, while the azithromycin sample in PBS (right tube) contained a large amount of crystals in the liquid and on the glass tube. The CER-001 sample with azithromycin (middle tube) was more turbid than the pure CER-001 sample, but contained significantly fewer azithromycin crystals than the azithromycin sample in PBS.

[0406] The CER-001 solution was mixed with spironolactone to provide a final concentration of 1.5 mg / ml of spironolactone and subjected to five heating and cooling cycles between 37°C and 55°C to promote the complexation of spironolactone with CER-001. For control, a CER-001 sample without spironolactone added and a sample of spironolactone at a concentration of 1.5 mg / ml in phosphate buffered saline (PBS) were also subjected to five heating and cooling cycles between 37°C and 55°C. Figure 1B As shown, the CER-001 sample without spironolactone (left tube) remained clear after the heating and cooling cycles, while the spironolactone sample in PBS (right tube) contained numerous crystals in the liquid, in the glass tube, and above the meniscus. The CER-001 sample with spironolactone (middle tube) was more turbid than CER-001 alone but contained significantly fewer spironolactone crystals than the spironolactone sample in PBS.

[0407] 7.1.2. Dexamethasone palmitate

[0408] Dexamethasone palmitate is a lipophilic prodrug of dexamethasone that can be used to treat macular edema (Daull et al., 2013, J. Ocul Pharmacol Ther. 29(2):258-69). A study was conducted to evaluate the suitability of CER-001 as a drug carrier to deliver dexamethasone palmitate.

[0409] The CER-001 solution was mixed with dexamethasone palmitate to provide a final concentration of 1 mg / ml of dexamethasone palmitate and subjected to five heating and cooling cycles between 37°C and 55°C to promote the complexation of dexamethasone palmitate with CER-001. For controls, CER-001 samples without dexamethasone palmitate added and samples of dexamethasone palmitate at a concentration of 1 mg / ml in phosphate buffered saline (PBS) were also subjected to five heating and cooling cycles between 37°C and 55°C. Figure 1C As shown, the CER-001 sample without dexamethasone palmitate (left tube) remained clear after the heating and cooling cycles, while the dexamethasone palmitate sample in PBS (right tube) contained a lipid film on the glass above the meniscus and was very cloudy or "milky." The CER-001 sample with dexamethasone palmitate (middle tube) was more cloudy than CER-001 alone, but contained no precipitate or crystals.

[0410] 7.1.3. Cyclosporine

[0411] Cyclosporine is an immunomodulator used to increase tear production in subjects with dry eye (Ames and Galor, 2015, Clin Investig (Longd.) 5(3):267-285). A study was conducted to evaluate the suitability of CER-001 as a drug carrier to deliver cyclosporine.

[0412] The CER-001 solution was mixed with cyclosporin to provide a final cyclosporin concentration of 1 mg / ml and subjected to five heating and cooling cycles between 37°C and 55°C to promote the complexation of cyclosporin with CER-001. For controls, a CER-001 sample without cyclosporin added and a sample of cyclosporin at a concentration of 1 mg / ml in phosphate buffered saline (PBS) were also subjected to five heating and cooling cycles between 37°C and 55°C. Figure 1D As shown, the CER-001 sample without cyclosporin (left tube) remained clear after the heating and cooling cycles, while the cyclosporin sample in PBS (right tube) contained crystals in the glass and liquid. The CER-001 sample with cyclosporin (middle tube) was more turbid than CER-001 alone, but contained no precipitate or crystals, even after overnight storage at 4°C.

[0413] This example shows that CER-001 can be complexed with azithromycin, spironolactone, dexamethasone palmitate, and cyclosporine, indicating that CER-001 is a suitable carrier for ophthalmic drugs.

[0414] 7.2. Example 2: Ocular distribution of CER-001 after intravenous administration in rabbits

[0415] A study was conducted to evaluate the distribution of CER-001 in the aqueous humor, vitreous, retina, and choroid of both eyes and in plasma following multiple intravenous (IV) administrations in albino rabbits. Briefly, on days 1, 3, and 5, each rabbit was administered a single IV injection of 2 mL / kg of CER-001 (16.2 mg / kg) at 8.1 mg / mL. On day 7, rabbits were administered two IV injections of 1.25 mL / kg of CER-001 (10.13 mg / kg per injection) at 8.1 mg / mL. Samples were taken 1 hour after the last IV administration for bioanalysis.

[0416] The greatest amount of CER-001 was detected in plasma (454 μg / mL). Smaller amounts of CER-001 were found in the choroid (16.4 μg / g), retina, and aqueous humor (2.1 μg / g and 2.7 μg / mL, respectively). CER-001 was not detected in the vitreous at the time of sampling.

[0417] The results of this example demonstrate that IV administration of CER-001 is associated with localization of CER-001 to ocular structures.

[0418] 7.3. Example 3: Ocular distribution of CER-001 after intravitreal administration in rabbits

[0419] A study was conducted to evaluate the distribution of CER-001 in the aqueous humor, vitreous, retina, and choroid of both eyes and in plasma following intravitreal (IVT) administration in albino rabbits. Briefly, a single IVT injection of 50 μL of CER-001 at 8.1 mg / mL was administered to the right eye of 6 albino rabbits on day 1, followed by a single IVT injection of 50 μL of CER-001 at 8.1 mg / mL to the left eye on day 2 (2 animals) or day 3 (the remaining 4 animals). Sampling for bioanalysis was performed 24, 48, or 120 hours after the last injection.

[0420] CER-001 was detected in all ocular tissues analyzed. The highest amount of CER-001 was found in the vitreous (approximately 140 μg / mL), which remained stable for 144 hours after injection. The amount of CER-001 in the aqueous humor (approximately 20 μg / mL), retina (approximately 40 μg / mL), and choroid (approximately 30 μg / g) was relatively low, but stable at all sampling time points. Since CER-001 was not detected in any plasma samples, no systemic exposure was observed. The complete results are shown in Table 2:

[0421]

[0422]

[0423] The results of this example demonstrate that this mode of administration is effective for localization of the CER-001 formulation in the ocular structures analyzed, particularly in the vitreous. For the IVT route of administration, the highest levels would be expected to be found in the vitreous. Distribution in all ocular tissues remained stable at all time points. No systemic exposure was observed.

[0424] 7.4. Example 4: CER-001 therapy for visual impairment associated with LCAT deficiency

[0425] CER-001 is administered to a subject having LCAT deficiency and having visual impairment associated with the subject's LCAT deficiency, said visual impairment being due to ocular lipid deposition, according to a treatment regimen comprising an induction regimen, a consolidation regimen, and a maintenance regimen.

[0426] Prior to treatment with CER-001, the subject had ocular lipid deposition manifested as white corneal ring opacities. The subject had normal vision but blurred vision, especially at night. Slit-lamp examination and optical computed tomography showed hyperreflective corneal opacities (data not shown). Next-generation sequencing confirmed that the subject was a compound heterozygote for two LCAT gene variants, but not in the ABCA1 or APOA1 genes. The first allele, maternally inherited, is a missense mutation p.(Ile202Thr) in exon 5 (c.605T>C), previously identified in Europe as familial LCAT deficiency (FLD). The paternal allele (c.154+5G>C) is novel and not present in general referral population databases. It alters highly evolutionarily conserved residues in the intron-1 donor splice site, thereby potentially altering exon-1 mRNA splicing and creating a cryptic acceptor splice site at position c.154+15. Therefore, if LCAT expression is not eliminated, mRNA incorporation of intronic sequences may produce abnormal / truncated proteins.

[0427] The induction regimen consisted of nine doses of CER-001 administered over three weeks. The dose of CER-001 administered in the induction regimen was 10 mg / kg, which was calculated based on the amount of ApoA-I in the administered CER-001 and the subject's body weight.

[0428] After the induction regimen, the subjects were administered CER-001 according to a consolidation regimen, which consisted of 7 doses of CER-001 administered over 4 weeks. The dose of CER-001 administered in the induction regimen was 10 mg / kg, which was calculated based on the amount of ApoA-I in the administered CER-001 and the subject's body weight.

[0429] After the consolidation regimen, the subjects were administered CER-001 according to a maintenance regimen, which included administration of CER-001 once a week for three weeks. The dose of CER-001 administered in the maintenance regimen was 10 mg / kg, which was calculated based on the amount of ApoA-I in CER-001 to be administered and the subject's body weight. Thereafter, the dose was increased to 20 mg / kg once a week for six weeks. The treatment period was 5 months, followed by a 3-month drug-free follow-up period.

[0430] CER-001 was administered as an IV infusion after premedication with hydroxyzine in the induction, consolidation, and maintenance regimens. CER-001 stock solution was diluted with saline (0.9% NaCl) prior to administration, and all doses of CER-001 were administered over one hour using an infusion pump at a fixed rate of 250 ml / hr.

[0431] During the CER-001 treatment, the subject's vision improved. In particular, the administration of CER-001 was accompanied by normalization of vision. At the end of the follow-up period, blurred vision did not recur.

[0432] CER-001 administered by infusion appears to have reached the anterior segment of the subject's eye, including the cornea, where it exerted its therapeutic effect. Without being bound by theory, it is believed that the observed effects on the subject's vision are due to the ability of CER-001, even when administered peripherally, to mobilize lipids accumulated in and / or around the eye (e.g., directly or indirectly). Additionally, also without being bound by theory, it is believed that the anti-inflammatory properties of CER-001 may contribute to the observed effects on the subject's vision.

[0433] It is further believed, again without being bound by theory, that subjects with other eye diseases, particularly those associated with lipid accumulation, may similarly benefit from treatment with CER-001 or another lipid binding protein-based complex. In addition, again without being bound by theory, it is believed that the ability of CER-001 to reach the anterior segment of the eye when administered peripherally can be used to deliver ophthalmic drugs to the eye (e.g., the anterior segment of the eye).

[0434] 7.5. Example 5: Ocular Tolerance of CER-001 in Rabbits

[0435] The ocular tolerability of CER-001 was evaluated in albino rabbits. In the first evaluation, CER-001 with or without complexed dexamethasone palmitate was administered to the eyes of albino rabbits at 8 mg / mL (based on protein weight), topically or by a single IVT injection. No tolerability issues were observed with repeated topical applications of up to 8 drops or a single intravitreal administration.

[0436] In the second evaluation, the ocular tolerability of CER-001 with or without softcorticosteroid (S-Cort) was assessed following repeated daily instillations of 50 μL or a single IVT injection of 50 μL into the right eye. As was the case with the first evaluation, no tolerability concerns were observed in any of the test groups, indicating that CER-001 and CER-001 with S-Cort were well tolerated macroscopically.

[0437] 7.6. Example 6: CER-001 treatment of endotoxin-induced uveitis (severe inflammation) in rabbits

[0438] A study was conducted to evaluate the ability of CER-001 to treat endotoxin-induced uveitis (severe inflammation) in albino rabbits when administered topically or by a single intravitreal injection (IVT) with or without complexed dexamethasone palmitate (DXP). An injectable formulation containing the anti-inflammatory glucocorticoid methylprednisolone sodium succinate was used as a control. Tolerance was assessed by the McDonald-Shadduck scoring system (see Eaton et al., Journal of Ocular Pharmacology and Therapeutics 33(10):718-734) at 6 and 24 hours after administration. Cell infiltration and protein content in aqueous humor were measured 24 hours after administration.

[0439] Briefly, the first study used five groups of 10 animals:

[0440] Group 1: Vehicle was topically applied to the right eye.

[0441] Group 2: 8 mg / ml of CER-001 was applied to the right eye surface (3 times of 50 μl).

[0442] • Group 3: Vehicle was administered IVT in the right eye.

[0443] • Group 4: CER-001 at 0.8 mg / ml was administered in 50 μl IVT in the right eye.

[0444] Group 5: Solu-Medrol 20 mg / ml was administered subconjunctivally in 250 μl in the right eye (as a positive control).

[0445] Add a group at one month intervals from the previous life stage.

[0446] • Group 6: 8 mg / ml CER-001 was administered in 50 μl IVT in the right eye.

[0447] Add another group at a gap of one month from the previous life stage.

[0448] Group 7: 8 mg / ml CER-S Cort was administered in 50 μl IVT in the right eye. CER-S Cort (soft corticosteroid) was prepared using 8 mg / ml CER-001.

[0449] 7.6.1. Methods

[0450] The right eye of each rabbit was examined using a slit lamp.

[0451] Observations of the conjunctiva (redness and swelling), aqueous flare, and iris were scored using a modified McDonald-Shadduck as follows:

[0452] Modified McDonald and Shadduck scale

[0453] conjunctiva

[0454]

[0455]

[0456] Aqueous Flash

[0457] The intensity of the Tyndall phenomenon can be scored by comparing the normal Tyndall effect observed when the slit-lamp beam passes through the lens with the Tyndall effect observed in the anterior chamber. The presence of aqueous flare is presumptive evidence of a breakdown of the blood-aqueous barrier.

[0458]

[0459] a, The presence of hypopyon in the aqueous flare results in a maximum score of 3.

[0460] IRIS

[0461] In the following definitions, primary, secondary, and tertiary vessels are used to help determine the subjective ocular score of iris involvement. It is assumed that the more vascular congestion there is, the more secondary and tertiary vessel involvement there is, and the greater the intensity of iris involvement. Scores range from 0 to +4.

[0462]

[0463] bThe presence of iris synechiae results in a maximum score of 4.

[0464] Tolerability of different treatment groups Figures 2A-2C Data are presented as mean ± SEM. One-way ANOVA test results: *: p < 0.05; **: p < 0.01; ****: p < 0.0001.

[0465] After Giemsa staining under a microscope, histological observation was performed to determine the number of infiltrating cells in the aqueous humor (AH). Figure 3A Data are presented as mean ± SEM. One-way ANOVA results: *: p < 0.05; **: p < 0.01; ****: p < 0.0001.

[0466] The protein concentration in AH was measured by protein-dye binding assay (Bradford test). Figure 3B Data are presented as mean ± SEM. One-way ANOVA test results: *: p < 0.05; **: p < 0.01; ****: p < 0.0001.

[0467] For ocular examination, the total grade consisted of the sum of the scores in each of the three categories: conjunctiva (redness, swelling), aqueous flare, and iris. Group means and standard deviations or medians were calculated for AH leukocyte density, AH protein concentration, cumulative ocular clinical scores, and body weight.

[0468] Transient opacity was observed in Group 7 (CER-S Cort). The opacity scores ranged from 1 to 3 and were defined as follows:

[0469] 1. The test article is transparent, but may produce refractive deviation when observing the underlying fundus structures.

[0470] 2. The test article is translucent, allowing only a vague view of the underlying fundus structures.

[0471] 3. The test article is opaque, obscuring the observation of the underlying fundus structures.

[0472] Figure 4 The turbidity results of group 7 at 0, 6 and 24 hours after induction are shown. Data are expressed as mean ± SEM. One-way ANOVA test *: p < 0.05; **: p < 0.01; ****: p < 0.0001.

[0473] Tolerability of different treatment groups Figures 2A-2C As shown in Figure 2, while aqueous humor cell infiltration and protein content are shown in Figure 2 Figure 3A-3B CER-001 induced significant tolerability in a single intravitreal administration (including a high dose of 8 mg / ml) with or without dexamethasone palmitate ( Figures 2A-2C ). Positive effects were similarly observed on cellular infiltration and proteins in the aqueous humor ( Figure 3A-3B ). This example further supports the use of CER-001 and similar lipid binding protein-based complexes to treat eye diseases such as uveitis, and the use of CER-001 and similar lipid binding protein-based complexes to deliver ophthalmic drugs to the eye to treat eye diseases such as uveitis.

[0474] 7.7. Example 7: Efficacy of a single IVT administration of CER-001 in a rabbit model of DL-α-aminoadipic acid-induced retinal neovascularization

[0475] The efficacy of CER-001 on retinal DL-α-aminoadipate (DL-AAA)-induced neovascularization and permeability was evaluated in pigmented rabbits following a single IVT administration. DL-α-aminoadipate (AAA) is a retinal glial cell toxin known to cause glial dysfunction and death, resulting in blood-retinal barrier breakdown and persistent retinal neovascularization for 10-12 weeks.

[0476] Briefly, pigmented rabbits were randomly divided into 3 groups: 8 mg / mL CER-001 group (n=6), 15 mg / mL CER-001 group (n=4) and vehicle (n=5). 12 weeks before the start of CER-001 treatment, 50 μL DL-AAA was administered by IVT in the right eye of all rabbits to induce neovascularization.

[0477] Before CER-001 or vehicle was administered to the right eye of each rabbit via IVT, a baseline Heidelberg retinal angiography (HRA) was assessed. After CER-001 or vehicle was administered on day 0, HRA was assessed weekly. Laser flare meter (LFM) and intraocular pressure (IOP) were assessed in the CER-001 treatment group on day 25, and laser flare meter (LFM) and intraocular pressure (IOP) were assessed in the vehicle-treated group on day 18. IOP is used to assess treatment-related changes in intraocular fluid pressure, and LFM is used to assess intraocular inflammation. For the CER-001 treatment group, rabbits were euthanized and tissues were sampled on day 40, and for the vehicle-treated group, rabbits were euthanized and tissues were sampled on day 33.

[0478] Fluorescein angiography HRA results were used to evaluate vascular leakage. Both CER-001 treatment groups were associated with reduced vascular leakage, corresponding to reduced neovascularization and permeability (Table 3). In all HRA assessments, floating bodies were observed in the vitreous of both CER-001 treated rabbits but not in vehicle treated rabbits.

[0479]

[0480]

[0481] The intraocular pressure (IOP) values ​​were comparable between the left (control) and right (treated) eyes of rabbits in the CER-001 (8 mg / mL) and vehicle groups. A slight (<20%) decrease in IOP was observed in the right eyes of 50% of the rabbits in the CER-001 (15 mg / mL) group (data not shown). Similarly, the laser flare meter (LFM) values ​​of rabbits treated with CER-001 (15 mg / mL) were elevated relative to those observed in the low-dose CER-001 and vehicle groups, as shown in Table 4.

[0482]

[0483] The results of this example demonstrate that CER-001 can reduce retinal neovascularization and permeability.

[0484] 7.8. Example 8: Efficacy and ocular tolerability of CER-001 in a rabbit model of retinal vascular hyperpermeability

[0485] The efficacy and ocular tolerability of different doses of CER-001 on retinal vascular hyperpermeability were evaluated after a single IVT administration. Briefly, 50 μL of CER-001 (1, 2, 4, or 6 mg / mL), aflibercept (40 mg / mL), or vehicle was administered by IVT into the eyes of pigmented rabbits 2 days prior to VEGF administration via IVT. Group 1, n=5, right eyes received 6 mg / mL CER-001 and left eyes received 4 mg / mL CER-001. Group 2, n=5, right eyes received 2 mg / mL CER-001 and left eyes received 1 mg / mL CER-001. Group 3, n=5, right eyes received vehicle and left eyes received aflibercept. Vascular leakage was assessed via fluorophotometry on day 2 after VEGF administration and compared to baseline values ​​determined before day -2. Tolerability was assessed by the McDonald-Shadduck scoring system on Day -2 (administration of CER-001, aflibercept or vehicle), Day 0 (administration of VEGF) and Day 2 (vascular leakage assessment). The results of the vascular leakage assessment are presented in Table 5 as the ratio of the area under the curve (AUC) of the vitreoretinal segment at Day 2 to baseline.

[0486]

[0487]

[0488] Fluorescence photometry results showed that administration of the positive control aflibercept resulted in the lowest vitreoretinal fluorescence ratio. In addition, all CER-001-treated groups had lower vitreoretinal fluorescence ratios relative to the vehicle group. Administration of CER-001 was associated with floating bodies in the vitreous. McDonald-Shadduck assessment did not reveal any major clinical symptoms after administration of any CER-001 dose. In summary, these results suggest that CER-001 can reduce VEGF-induced retinal vascular permeability.

[0489] In a second evaluation, the efficacy of CER-001 in a rabbit model of retinal vascular hyperpermeability was evaluated after multiple IVT administrations. This time, 50 μL of CER-001 (8.1 mg / mL) or vehicle was administered by IVT into the right eye of pigmented rabbits 3 days before and 1 day after VEGF administration via IVT. Vascular leakage was assessed via fluorophotometry on day 2 after VEGF administration.

[0490] The vitreoretinal fluorescence ratio of the treated / untreated eyes was lower in the CER-001 treated group relative to the vehicle group on day 2. Specifically, the mean ratio was 14±9 in the CER-001 treated group and 31±18 in the vehicle group. These results suggest that CER-001 administered via IVT can reduce VEGF-induced retinal vascular permeability.

[0491] 7.9. Example 9: Efficacy of CER-001 treatment against chronic panuveitis caused by Mycobacterium tuberculosis in rabbits

[0492] The efficacy of multiple IV administrations of CER-001 against uveitis caused by Mycobacterium tuberculosis was evaluated in albino rabbits. Briefly, each rabbit was injected subcutaneously (SC) with H37Ra on day 1 to sensitize the rabbits to Mycobacterium tuberculosis antigens. Uveitis induction involved IVT injections of H37Ra on days 15 and 30. Rabbits were randomly assigned to treatment groups of CER-001 (8.1 mg / mL) (n=4) or vehicle (n=4). IV treatments of CER-001 (2 mL / kg, 16.2 mg / kg) or vehicle were performed on days 1, 5, 9, 13, 17, 21, 25, and 29. Ocular examinations were performed using a slit lamp to assess ocular inflammation in the anterior and posterior ocular regions.

[0493] IVT injection of H37Ra antigen resulted in panuveitis in both the anterior and posterior parts of the eyes of vehicle-treated rabbits. For example, anterior and posterior ocular inflammation peaked 72 hours after each IVT injection. Rabbits in the CER-001-treated group had less pronounced levels of anterior and posterior ocular inflammation after the first IVT injection. There was no difference in posterior ocular inflammation scores between the two treatment groups after the second IVT injection. The results of the slit lamp evaluations are summarized in Table 6 below.

[0494]

[0495] 8. Specific implementation plan

[0496] Various aspects of the present disclosure are described in the embodiments set forth in the following numbered paragraphs of Group 1.

[0497] Group 1:

[0498] 1. A method of treating a subject suffering from an ocular disease, comprising administering to the subject an amount of a lipid binding protein-based complex, optionally in combination with one or more ophthalmic drugs, effective to reduce the severity of the ocular disease, wherein:

[0499] (a) the subject suffers from cataract or acute anterior uveitis; and / or

[0500] (b) The lipid binding protein-based complex is administered by suprachoroidal injection, suprascleral injection, implantation, iontophoresis or electroporation.

[0501] 2. The method of embodiment 1, wherein the eye disease is a disease associated with lipid accumulation.

[0502] 3. The method of embodiment 2, wherein the eye disease is fish eye disease.

[0503] 4. The method of embodiment 2, wherein the eye disease is lipid keratopathy.

[0504] 5. The method of embodiment 4, wherein the lipid keratopathy is secondary lipid keratopathy.

[0505] 6. The method of embodiment 2, wherein the eye disease is a corneal dystrophy, such as a hereditary corneal dystrophy, anterior or superficial corneal dystrophy, stromal corneal dystrophy, or posterior corneal dystrophy.

[0506] 7. The method of any one of embodiments 1 to 6, wherein the subject suffers from corneal opacity and wherein the amount of the lipid binding protein-based complex is an amount effective to reduce corneal opacity in the subject.

[0507] 8. The method of embodiment 7, wherein opacity is measured by anterior segment optical coherence tomography (OCT).

[0508] 9. The method of any one of embodiments 1 to 8, wherein the amount of the lipid binding protein-based complex is an amount effective to improve contrast sensitivity in the subject.

[0509] 10. The method of any one of embodiments 1 to 9, wherein the amount of the lipid binding protein-based complex is an amount effective to reduce the stray light value of the subject.

[0510] 11. The method of any one of embodiments 1 to 10, wherein the subject is homozygous for the LCAT mutation.

[0511] 12. The method of any one of embodiments 1 to 10, wherein the subject is heterozygous for the LCAT mutation.

[0512] 13. The method of any one of embodiments 1 to 2 and 4 to 10, unless dependent upon embodiment 3, wherein the subject does not have LCAT deficiency.

[0513] 14. The method of embodiment 1 or embodiment 2, wherein the subject suffers from cataracts.

[0514] 15. The method of embodiment 1 or embodiment 2, wherein the eye disease is dry eye.

[0515] 16. The method of embodiment 15, wherein the dry eye is associated with meibomian gland dysfunction (MGD).

[0516] 17. The method of embodiment 16, wherein the MGD is obstructive MGD.

[0517] 18. The method of embodiment 15, wherein the dry eye is associated with tear gland dysfunction.

[0518] 19. The method of embodiment 1 or embodiment 2, wherein the eye disease is blepharitis.

[0519] 20. The method of embodiment 1 or embodiment 2, wherein the eye disease is an inflammatory eye disease.

[0520] 21. The method of embodiment 1 or embodiment 2, wherein the eye disease is uveitis.

[0521] 22. The method of embodiment 21, wherein the uveitis is anterior uveitis, intermediate uveitis, posterior uveitis or panuveitis.

[0522] 23. The method of embodiment 22, wherein the uveitis is anterior uveitis.

[0523] 24. The method of claim 23, wherein the uveitis is acute anterior uveitis.

[0524] 25. The method of embodiment 22, wherein the uveitis is intermediate uveitis.

[0525] 26. The method of embodiment 22, wherein the uveitis is posterior uveitis.

[0526] 27. The method of embodiment 22, wherein the uveitis is panuveitis.

[0527] 28. The method according to any one of embodiments 21 to 27, wherein the uveitis is caused by a bacterial infection.

[0528] 29. The method of embodiment 1 or embodiment 2, wherein the eye disease is macular edema, macular degeneration, retinal detachment, eye tumor, fungal infection, viral infection, bacterial infection (e.g., bacterial conjunctivitis or trachoma), multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis, or glaucoma.

[0529] 30. The method of embodiment 1 or embodiment 2, wherein the eye disease is dry form of macular degeneration.

[0530] 31. The method of embodiment 1 or embodiment 2, wherein the eye disease is wet macular degeneration.

[0531] 32. The method of embodiment 1 or embodiment 2, wherein the eye disease is diabetic retinopathy, optionally wherein the subject has diabetic macular edema.

[0532] 33. The method of embodiment 1 or embodiment 2, wherein the eye disease is Stargardt's disease.

[0533] 34. The method of any one of embodiments 1 to 33, wherein the subject has impaired vision due to an eye disease, and the amount of the lipid binding protein-based complex is an amount that improves the subject's vision.

[0534] 35. The method of any one of embodiments 1 to 34, wherein the subject suffers from ocular lipid deposition.

[0535] 36. The method of embodiment 35, wherein the ocular lipid deposition comprises corneal lipid deposition, retinal lipid deposition, eyelid lipid deposition, or a combination thereof.

[0536] 37. The method of embodiment 36, wherein the ocular lipid deposition comprises corneal lipid deposition.

[0537] 38. The method of embodiment 36 or embodiment 37, wherein the ocular lipid deposits comprise retinal lipid deposits.

[0538] 39. The method of any one of embodiments 36 to 38, wherein the ocular lipid deposition comprises eyelid lipid deposition.

[0539] 40. The method of any one of embodiments 35 to 39, wherein the ocular lipid deposits are not calcified.

[0540] 41. The method of any one of embodiments 35 to 40, wherein the lipid deposits comprise lipid deposits within drusen deposits.

[0541] 42. The method of any one of embodiments 35 to 40, wherein the lipid deposits comprise lipofuscin particles.

[0542] 43. The method of any one of embodiments 35 to 40, wherein the lipid deposits comprise cholesterol deposits.

[0543] 44. The method of any one of embodiments 35 to 43, comprising administering to the subject an amount of the lipid binding protein-based complex effective to reduce the size and / or number of ocular lipid deposits.

[0544] 45. The method of any one of embodiments 1 to 44, wherein the lipid binding protein-based complex comprises apolipoprotein AI (ApoA-I), optionally wherein the ApoA-I is not ApoA-I Milano .

[0545] 46. ​​The method of any one of embodiments 1 to 45, wherein the lipid binding protein-based complex does not comprise an apolipoprotein mimetic.

[0546] 47. The method of any one of embodiments 1 to 46, wherein the lipid binding protein-based complex is a remodeled HDL or a HDL mimetic.

[0547] 48. The method of embodiment 47, wherein the lipid binding protein-based complex comprises CER-001.

[0548] 49. The method of embodiment 48, wherein the CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids, the ApoA-I weight:total phospholipids weight ratio being 1:2.7+ / -20%, and the phospholipids sphingomyelin and DPPG, the sphingomyelin:DPPG weight:weight ratio being 97:3+ / -20%.

[0549] 50. The method of embodiment 48, wherein the CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids, the ApoA-I weight:total phospholipids weight ratio being 1:2.7+ / -10%, and the phospholipids sphingomyelin and DPPG, the sphingomyelin:DPPG weight:weight ratio being 97:3+ / -10%.

[0550] 51. The method of embodiment 48, wherein the CER-001 is a lipoprotein complex comprising ApoA-I and phospholipids, with an ApoA-I weight:total phospholipid weight ratio of 1:2.7, and the phospholipids sphingomyelin and DPPG, with a sphingomyelin:DPPG weight:weight ratio of 97:3.

[0551] 52. The method of any one of embodiments 49 to 51, wherein the ApoA-I has the amino acid sequence of amino acids 25-267 of SEQ ID NO: 1 of WO 2012 / 109162.

[0552] 53. The method of any one of embodiments 49 to 52, wherein the ApoA-I is recombinantly expressed.

[0553] 54. The method of any one of embodiments 49 to 53, wherein the CER-001 comprises a native sphingomyelin.

[0554] 55. The method of embodiment 54, wherein the natural sphingomyelin is hen egg sphingomyelin.

[0555] 56. The method of any one of embodiments 49 to 53, wherein the CER-001 comprises a synthetic sphingomyelin.

[0556] 57. The method of embodiment 56, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin.

[0557] 58. The method of any one of embodiments 48 to 57, wherein CER-001 is administered in the form of a formulation, wherein the CER-001 is at least 95% homogeneous.

[0558] 59. The method of embodiment 58, wherein CER-001 is administered in the form of a formulation, wherein the CER-001 is at least 97% homogeneous.

[0559] 60. The method of embodiment 58, wherein CER-001 is administered in the form of a formulation, wherein the CER-001 is at least 98% homogeneous.

[0560] 61. The method of embodiment 58, wherein CER-001 is administered in the form of a formulation, wherein the CER-001 is at least 99% homogeneous.

[0561] 62. The method of embodiment 47, wherein the lipid binding protein-based complex comprises CSL-111.

[0562] 63. The method of embodiment 47, wherein the lipid binding protein-based complex comprises CSL-112.

[0563] 64. The method of embodiment 47, wherein the lipid binding protein-based complex comprises ETC-216.

[0564] 65. The method of embodiment 47, wherein the lipid binding protein-based complex comprises CER-522.

[0565] 66. The method of embodiment 47, wherein the lipid binding protein-based complex comprises delipidated HDL.

[0566] 67. The method of any one of embodiments 1 to 45, wherein the lipid binding protein-based complex is a cargomer.

[0567] 68. The method of any one of embodiments 1 to 67, wherein the lipid binding protein-based complex is a carrier for one or more ophthalmic drugs, optionally wherein one or more of the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

[0568] 69. The method of any one of embodiments 1 to 68, wherein the lipid-binding protein-based complex comprises a lipid-binding protein-based complex having one or more ophthalmic drugs complexed therewith, optionally wherein one or more of the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

[0569] 70. The method of embodiment 68 or embodiment 69, wherein the one or more ophthalmic drugs comprise a steroid, a kinase inhibitor, an angiotensin II receptor antagonist, an aldose reductase inhibitor, an immunosuppressant, a carbonic anhydrase inhibitor, an antimicrobial agent, an antiviral agent, an antihistamine, an anti-inflammatory agent, a prostaglandin analog, or a combination thereof.

[0570] 71. The method of any one of embodiments 68 to 70, wherein the one or more ophthalmic drugs include azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyridinol [2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, 2-methylsorbitol, cypermethrin Rolimus, cyclosporine, tacrolimus, acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, latanoprost, trovaprost, bimatoprost, or a combination thereof.

[0571] 72. The method of any one of embodiments 68 to 70, wherein the one or more ophthalmic drugs include azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyrido[2,3 -d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, 2-methylsorbitol, sirolimus, cyclosporine tacrolimus, acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, latanoprost, trovaprost, bimatoprost, dexamethasone palmitate, or a combination thereof.

[0572] 73. The method of any one of embodiments 68 to 72, wherein the one or more ophthalmic drugs comprises azithromycin.

[0573] 74. The method of any one of embodiments 68 to 73, wherein the one or more ophthalmic drugs comprises spironolactone.

[0574] 75. The method of any one of embodiments 68 to 74, wherein the one or more ophthalmic drugs comprises dexamethasone palmitate.

[0575] 76. The method of any one of embodiments 68 to 75, wherein the one or more ophthalmic drugs comprises cyclosporine.

[0576] 77. The method of any one of embodiments 68 to 76, wherein the one or more ophthalmic drugs comprises latanoprost, trovaprost, bimatoprost, tafluprost, or a combination thereof.

[0577] 78. The method of embodiment 77, wherein the one or more ophthalmic drugs comprises latanoprost.

[0578] 79. The method of any one of embodiments 68 to 78, wherein the one or more ophthalmic drugs comprises dexamethasone.

[0579] 80. The method of any one of embodiments 68 to 79, wherein the one or more ophthalmic drugs comprises loteprednol etabonate.

[0580] 81. The method of any one of embodiments 68 to 80, wherein the one or more ophthalmic drugs comprises triamcinolone acetonide.

[0581] 82. The method of any one of embodiments 68 to 81, wherein the one or more ophthalmic drugs comprises acyclovir.

[0582] 83. The method of any one of embodiments 68 to 82, wherein the one or more ophthalmic drugs comprises trovaprost.

[0583] 84. The method of any one of embodiments 68 to 83, wherein the one or more ophthalmic drugs comprises bimatoprost.

[0584] 85. The method of any one of embodiments 68 to 84, wherein the one or more ophthalmic drugs comprises tafluprost.

[0585] 86. The method of any one of embodiments 68 to 85, wherein the one or more ophthalmic drugs comprises pazopanib.

[0586] 87. The method of any one of embodiments 68 to 86, wherein the one or more ophthalmic drugs comprises sirolimus.

[0587] 88. The method of any one of embodiments 68 to 86, wherein the one or more ophthalmic drugs comprises tacrolimus.

[0588] 89. The method of any one of embodiments 68 to 86, wherein the one or more ophthalmic drugs comprises nepafenac.

[0589] 90. The method of any one of embodiments 1 to 46, wherein the lipid binding protein-based complex is an apomer.

[0590] 91. The method of any one of embodiments 1 to 90, wherein the lipid binding protein-based complex is administered peripherally, optionally by infusion.

[0591] 92. The method of embodiment 91, wherein the lipid binding protein-based complex is administered according to a dosing regimen comprising:

[0592] (a) induction regimen; and / or

[0593] (b) a consolidation programme; and / or

[0594] (c) maintenance program,

[0595] Optionally, wherein the lipid binding protein-based complex comprises CER-001.

[0596] 93. The method of embodiment 92, comprising administering one or more doses of the lipid binding protein-based complex according to an induction regimen.

[0597] 94. The method of embodiment 93, wherein the induction regimen comprises administering multiple doses of the lipid binding protein-based complex to the subject.

[0598] 95. The method of embodiment 94, wherein the induction regimen comprises administering at least three doses of the lipid binding protein-based complex to the subject.

[0599] 96. The method of embodiment 94 or embodiment 95, wherein multiple doses in the induction regimen are separated by 1 day or more.

[0600] 97. The method of any one of embodiments 93 to 96, wherein the interval between doses after the initial dose of the induction regimen does not exceed 3 days.

[0601] 98. The method of embodiment 97, wherein the interval between doses after the initial dose of the induction regimen is one to three days.

[0602] 99. The method of embodiment 97, wherein the dose interval after the initial dose of the induction regimen is two to three days.

[0603] 100. The method of embodiment 97, wherein the dose intervals after the initial dose of the induction regimen are one to two days.

[0604] 101. The method of any one of embodiments 93 to 100, wherein the induction regimen lasts for at least one week.

[0605] 102. The method of embodiment 101, wherein the induction regimen lasts for two weeks.

[0606] 103. The method of embodiment 101, wherein the induction regimen lasts for three weeks.

[0607] 104. The method of any one of embodiments 93 to 103, wherein the induction regimen comprises administering three doses of the lipid binding protein-based complex to the subject per week.

[0608] 105. The method of any one of embodiments 93 to 103, wherein the induction regimen comprises administering four or more doses of the lipid binding protein-based complex to the subject.

[0609] 106. The method of any one of embodiments 93 to 103, wherein the induction regimen comprises administering five or more doses of the lipid binding protein-based complex to the subject.

[0610] 107. The method of any one of embodiments 93 to 103, wherein the induction regimen comprises administering six or more doses of the lipid binding protein-based complex to the subject.

[0611] 108. The method of any one of embodiments 93 to 103, wherein the induction regimen comprises administering seven or more doses of the lipid binding protein-based complex to the subject.

[0612] 109. The method of any one of embodiments 93 to 103, wherein the induction regimen comprises administering eight or more doses of the lipid binding protein-based complex to the subject.

[0613] 110. The method of any one of embodiments 93 to 103, wherein the induction regimen comprises administering nine or more doses of the lipid binding protein-based complex to the subject.

[0614] 111. The method of embodiment 110, wherein the induction regimen comprises administering a first dose of the lipid binding protein-based complex to the subject on day 1 and administering subsequent doses of the induction regimen to the subject on days 2, 4, 7, 9, 11, 14, 16 and 18.

[0615] 112. The method of any one of embodiments 93 to 103, wherein the induction regimen comprises administering ten or more doses of the lipid binding protein-based complex to the subject.

[0616] 113. The method of embodiment 92, which does not include an induction protocol.

[0617] 114. The method of any one of embodiments 92 to 113, comprising administering one or more doses of the lipid binding protein-based complex to the subject according to a consolidation schedule.

[0618] 115. The method of embodiment 114, wherein the consolidation regimen comprises administering multiple doses of the lipid binding protein-based complex to the subject.

[0619] 116. The method of embodiment 115, wherein the multiple doses in the consolidation regimen are separated by 2 or more days.

[0620] 117. The method of any one of embodiments 114 to 116, wherein the consolidation regimen comprises administering to the subject at least two doses of the lipid binding protein-based complex within one week.

[0621] 118. The method of any one of embodiments 114 to 117, wherein the dosage intervals of the consolidation regimen do not exceed four days.

[0622] 119. The method of any one of embodiments 114 to 118, wherein the doses of the consolidation regimen are separated from each other by three or four days.

[0623] 120. The method according to any one of embodiments 114 to 119, wherein the consolidation regimen lasts for at least 3 weeks.

[0624] 121. The method of any one of embodiments 114 to 120, wherein the consolidation regimen comprises administering three or more doses of the lipid binding protein-based complex to the subject.

[0625] 122. The method of any one of embodiments 114 to 120, wherein the consolidation regimen comprises administering four or more doses of the lipid binding protein-based complex to the subject.

[0626] 123. The method of any one of embodiments 114 to 120, wherein the consolidation regimen comprises administering five or more doses of the lipid binding protein-based complex to the subject.

[0627] 124. The method of any one of embodiments 114 to 120, wherein the consolidation regimen comprises administering six or more doses of the lipid binding protein-based complex to the subject.

[0628] 125. The method of embodiment 124, wherein the consolidation regimen comprises administering six doses of the lipid binding protein-based complex to the subject.

[0629] 126. The method of embodiment 125, wherein the consolidation regimen comprises administering six doses of the lipid binding protein-based complex to the subject on days 21, 24, 28, 31, 35 and 38 following the induction regimen beginning on day 1.

[0630] 127. The method of any one of embodiments 114 to 120, wherein the consolidation regimen comprises administering seven or more doses of the lipid binding protein-based complex to the subject.

[0631] 128. The method of any one of embodiments 114 to 120, wherein the consolidation regimen comprises administering eight or more doses of the lipid binding protein-based complex to the subject.

[0632] 129. The method of any one of embodiments 114 to 120, wherein the consolidation regimen comprises administering nine or more doses of the lipid binding protein-based complex to the subject.

[0633] 130. The method of any one of embodiments 114 to 120, wherein the consolidation regimen comprises administering ten or more doses of the lipid binding protein-based complex to the subject.

[0634] 131. The method of any one of embodiments 92 to 113, which does not include a consolidation regimen.

[0635] 132. The method of any one of embodiments 92 to 131, comprising administering multiple doses of the lipid binding protein-based complex to the subject according to a maintenance regimen.

[0636] 133. The method of embodiment 132, wherein the maintenance regimen comprises administering a dose of the lipid binding protein-based complex to the subject every 3 days or more.

[0637] 134. The method of embodiment 132, wherein the maintenance regimen comprises administering a dose of the lipid binding protein-based complex to the subject every 5 days or more.

[0638] 135. The method of embodiment 132, wherein the maintenance regimen comprises administering a dose of the lipid binding protein-based complex to the subject every week.

[0639] 136. The method of embodiment 135, wherein the dosage of the maintenance regimen is administered + / - 2 days before and after a strict weekly date.

[0640] 137. The method of embodiment 132, wherein the maintenance regimen comprises administering a dose of the lipid binding protein-based complex to the subject twice per week.

[0641] 138. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for at least one month.

[0642] 139. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for at least two months.

[0643] 140. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for at least three months.

[0644] 141. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for at least six months.

[0645] 142. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for at least nine months.

[0646] 143. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for at least one year.

[0647] 144. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for at least 18 months.

[0648] 145. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for at least 2 years.

[0649] 146. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject indefinitely.

[0650] 147. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for 16 weeks or more.

[0651] 148. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for 20 weeks or more.

[0652] 149. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for 30 weeks or more.

[0653] 150. The method of any one of embodiments 132 to 137, wherein the maintenance regimen comprises administering the lipid binding protein-based complex to the subject for 40 weeks or more.

[0654] 151. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 4 to 30 mg / kg (based on protein weight).

[0655] 152. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 5 to 15 mg / kg (based on protein weight).

[0656] 153. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 10 to 20 mg / kg (based on protein weight).

[0657] 154. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 15 to 25 mg / kg (based on protein weight).

[0658] 155. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 8 mg / kg (based on protein weight).

[0659] 156. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 10 mg / kg (based on protein weight).

[0660] 157. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 300 mg to 3000 mg.

[0661] 158. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 300 mg to 1500 mg.

[0662] 159. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 400 mg to 1500 mg.

[0663] 160. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 500 mg to 1200 mg.

[0664] 161. The method of any one of embodiments 92 to 150, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is 500 mg to 1000 mg.

[0665] 162. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 4 to 30 mg / kg (based on protein weight).

[0666] 163. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 5 to 15 mg / kg (based on protein weight).

[0667] 164. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 10 to 20 mg / kg (based on protein weight).

[0668] 165. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 15 to 25 mg / kg (based on protein weight).

[0669] 166. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 8 mg / kg (based on protein weight).

[0670] 167. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 10 mg / kg (based on protein weight).

[0671] 168. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 300 mg to 3000 mg.

[0672] 169. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 300 mg to 1500 mg.

[0673] 170. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 400 mg to 1500 mg.

[0674] 171. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 500 mg to 1200 mg.

[0675] 172. The method of any one of embodiments 92 to 161, wherein the dose of the lipid binding protein-based complex administered in the consolidation regimen is 500 mg to 1000 mg.

[0676] 173. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 4 to 30 mg / kg (based on protein weight).

[0677] 174. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 5 to 15 mg / kg (based on protein weight).

[0678] 175. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 10 to 20 mg / kg (based on protein weight).

[0679] 176. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 15 to 25 mg / kg (based on protein weight).

[0680] 177. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 8 mg / kg (based on protein weight).

[0681] 178. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 10 mg / kg (based on protein weight).

[0682] 179. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 20 mg / kg (based on protein weight).

[0683] 180. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 300 mg to 3000 mg.

[0684] 181. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 300 mg to 1500 mg.

[0685] 182. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 400 mg to 1500 mg.

[0686] 183. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 500 mg to 1200 mg.

[0687] 184. The method of any one of embodiments 92 to 172, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 500 mg to 1000 mg.

[0688] 185. The method of any one of embodiments 92 to 184, comprising an induction regimen and a maintenance regimen.

[0689] 186. The method of embodiment 185, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is the same as the dose of the lipid binding protein-based complex administered in the maintenance regimen.

[0690] 187. The method of embodiment 185, wherein the dose of the lipid binding protein-based complex administered in the induction regimen is different from the dose of the lipid binding protein-based complex administered in the maintenance regimen.

[0691] 188. The method of embodiment 187, wherein the dose of the lipid-binding protein-based complex administered in the maintenance regimen is greater than the dose of the lipid-binding protein-based complex administered in the induction regimen.

[0692] 189. The method of embodiment 188, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is 1.5 to 3 times the dose of the lipid binding protein-based complex administered in the induction regimen.

[0693] 190. The method of embodiment 188, wherein the dose of the lipid binding protein-based complex administered in the maintenance regimen is twice the dose administered in the induction regimen.

[0694] 191. The method of any one of embodiments 1 to 90, wherein the lipid binding protein-based complex is administered topically.

[0695] 192. The method of embodiment 191, wherein the lipid binding protein-based complex is administered intraocularly.

[0696] 193. The method of embodiment 192, wherein the lipid binding protein-based complex is administered by intraocular injection.

[0697] 194. The method of embodiment 193, wherein the intraocular injection is a suprachoroidal injection.

[0698] 195. The method of embodiment 193, wherein the intraocular injection is a suprascleral injection.

[0699] 196. The method of embodiment 193, wherein the intraocular injection is an intravitreal injection.

[0700] 197. The method of embodiment 193, wherein the intraocular injection is a subconjunctival injection.

[0701] 198. The method of embodiment 193, wherein the intraocular injection is a juxtabulbar injection.

[0702] 199. The method of embodiment 193, wherein the intraocular injection is a periorbital injection.

[0703] 200. The method of embodiment 193, wherein the intraocular injection is a retrobulbar injection.

[0704] 201. The method of embodiment 191 or embodiment 192, wherein the lipid binding protein-based complex is administered as an implant.

[0705] 202. The method of embodiment 201, wherein the implant is a disc.

[0706] 203. The method of embodiment 201, wherein the implant is a wafer.

[0707] 204. The method of embodiment 201, wherein the implant is a plug.

[0708] 205. The method of embodiment 201, wherein the implant is a rod.

[0709] 206. The method of embodiment 201, wherein the implant is a pellet.

[0710] 207. The method of any one of embodiments 201 to 206, wherein the implant is biodegradable.

[0711] 208. The method of any one of embodiments 201 to 206, wherein the implant is non-biodegradable.

[0712] 209. The method of embodiment 191, wherein the lipid binding protein-based complex is administered by iontophoresis.

[0713] 210. The method of embodiment 209, wherein the lipid binding protein-based complex is administered by transcorneal iontophoresis.

[0714] 211. The method of embodiment 209, wherein the lipid binding protein-based complex is administered by transscleral iontophoresis.

[0715] 212. The method of embodiment 191, wherein the lipid binding protein-based complex is administered by electroporation.

[0716] 213. The method of embodiment 191, wherein the lipid binding protein-based complex is topically administered.

[0717] 214. The method of embodiment 213, wherein the lipid binding protein-based complex is formulated as eye drops.

[0718] 215. The method of any one of embodiments 191 to 214, wherein the lipid binding protein-based complex is administered according to a dosing regimen comprising:

[0719] (a) induction regimen; and / or

[0720] (b) a consolidation programme; and / or

[0721] (c) maintenance program,

[0722] Optionally, wherein the lipid binding protein based complex is CER-001.

[0723] 216. The method of embodiment 215, comprising an induction regimen.

[0724] 217. The method of embodiment 215 or embodiment 216, comprising a consolidation regimen.

[0725] 218. The method of any one of embodiments 215 to 217, comprising a maintenance regimen.

[0726] 219. The method of any one of embodiments 1 to 218, wherein an antihistamine is administered prior to administering one or more doses of the lipid binding protein-based complex.

[0727] 220. The method of any one of embodiments 1 to 219, wherein the subject is also treated with a lipid-controlling drug.

[0728] 221. The method of embodiment 220, wherein the lipid control drug comprises a statin.

[0729] 222. The method of embodiment 221, wherein the statin is atorvastatin, rosuvastatin, simvastatin, fluvastatin, lovastatin or pravastatin.

[0730] 223. The method of any one of embodiments 220 to 222, wherein the lipid control drug comprises a cholesterol absorption inhibitor.

[0731] 224. The method of embodiment 223, wherein the cholesterol absorption inhibitor is ezetimibe.

[0732] 225. The method of any one of embodiments 220 to 224, wherein the lipid control drug comprises niacin.

[0733] 226. The method of any one of embodiments 220 to 225, wherein the lipid control drug comprises aspirin.

[0734] 227. The method of any one of embodiments 220 to 226, wherein the lipid-control drug comprises a proprotein convertase subtilisin / kexin type 9 (PCSK9) inhibitor.

[0735] 228. The method of embodiment 227, wherein the PCSK9 inhibitor is an antibody.

[0736] 229. The method of embodiment 228, wherein the antibody is alirocumab, bococizumab, evolocumab, 1D05-IgG2 or LY3015014.

[0737] 230. The method of embodiment 227, wherein the PCSK9 inhibitor is a RNAi therapeutic agent.

[0738] 231. The method of embodiment 230, wherein the RNAi therapeutic agent is ALN-PCSSC.

[0739] 232. The method of any one of embodiments 220 to 231, further comprising administering to the subject a therapeutically effective amount of a lipid-control drug.

[0740] 233. The method of any one of embodiments 1 to 232, wherein the subject is also treated with standard of care therapy for the eye disease.

[0741] 234. The method of embodiment 233 further comprises administering standard of care therapy to the subject.

[0742] 235. The method of any one of embodiments 1 to 234, wherein the subject is also treated with an antihypertensive drug, optionally wherein the antihypertensive drug comprises one, two or all three of amlodipine, urapidil and furosemide.

[0743] 236. The method of any one of embodiments 1 to 235, wherein the lipid binding protein-based complex does not comprise a cell penetrating peptide and is not administered together with a cell penetrating peptide.

[0744] 237. The method of any one of embodiments 1 to 236, wherein the lipid binding protein-based complex does not comprise a chemical permeation enhancer and is not administered together with a chemical permeation enhancer.

[0745] 238. The method of any one of embodiments 1 to 237, wherein the lipid binding protein-based complex does not comprise a cytophilic peptide and is not administered together with a cytophilic peptide.

[0746] 239. A composition comprising a lipid binding protein-based complex and one or more ophthalmic drugs, wherein the composition is produced by a method comprising thermally cycling a mixture comprising a lipid binding protein-based complex and one or more ophthalmic drugs, optionally wherein one or more of the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

[0747] 240. The composition of embodiment 239, wherein the thermal cycle comprises

[0748] (a) heating the mixture from a temperature within a first temperature range to a temperature within a second temperature range,

[0749] (b) cooling the mixture of (a) from a temperature within the second temperature range to a temperature within the first temperature range; and

[0750] (c) optionally repeating steps (a) and (b) at least once.

[0751] 241. The composition of embodiment 240, wherein the thermal cycle comprises repeating steps (a) and (b) once.

[0752] 242. The composition of embodiment 240, wherein the thermal cycle comprises repeating steps (a) and (b) twice.

[0753] 243. The composition of embodiment 240, wherein the thermal cycle comprises repeating steps (a) and (b) three times.

[0754] 244. The composition of embodiment 240, wherein the thermal cycle comprises repeating steps (a) and (b) four times.

[0755] 245. The composition of embodiment 240, wherein the thermal cycle comprises repeating steps (a) and (b) five times.

[0756] 246. The composition of any one of embodiments 240 to 245, wherein the first temperature range is 30°C to 45°C.

[0757] 247. The composition of embodiment 246, wherein the temperature within the first temperature range is 37°C.

[0758] 248. The composition of any one of embodiments 240 to 247, wherein the second temperature range is 50°C to 65°C.

[0759] 249. The composition of embodiment 248, wherein the temperature within the second temperature range is 55°C.

[0760] 250. The composition of any one of embodiments 239 to 249, wherein the thermal cycling comprises thermal cycling the mixture between 37°C and 55°C.

[0761] 251. A composition comprising a lipid binding protein-based complex and one or more ophthalmic drugs complexed therewith, optionally wherein one or more of the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

[0762] 252. The composition of any one of embodiments 239 to 251, wherein the lipid binding protein-based complex comprises a lipid binding protein molecule described in Section 6.1.4.

[0763] 253. The composition of any one of embodiments 239 to 251, wherein the lipid binding protein-based complex comprises apolipoprotein AI (ApoA-I), optionally wherein the ApoA-I is not ApoA-I Milano .

[0764] 254. The composition of any one of embodiments 239 to 253, wherein the lipid binding protein-based complex does not comprise an apolipoprotein mimetic.

[0765] 255. The composition of any one of embodiments 239 to 254, wherein the lipid binding protein-based complex comprises one or more amphiphilic molecules described in Section 6.1.5.

[0766] 256. The composition of any one of embodiments 239 to 255, wherein the lipid binding protein-based complex comprises one or more neutral lipids.

[0767] 257. The composition of embodiment 256, wherein the one or more neutral lipids comprise sphingomyelin.

[0768] 258. The composition of any one of embodiments 239 to 257, wherein the lipid binding protein-based complex comprises one or more negatively charged lipids.

[0769] 259. The composition of embodiment 258, wherein the one or more negatively charged lipids comprise 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol) (DPPG) or a salt thereof.

[0770] 260. The composition of any one of embodiments 239 to 259, wherein the lipid binding protein-based complex is a remodeled HDL or a HDL mimetic.

[0771] 261. The composition of embodiment 260, wherein the lipid binding protein-based complex is CER-001.

[0772] 262. The composition of embodiment 260, wherein the lipid binding protein-based complex is CSL-111.

[0773] 263. The composition of embodiment 260, wherein the lipid binding protein-based complex is CSL-112.

[0774] 264. The composition of embodiment 260, wherein the lipid binding protein-based complex is ETC-216.

[0775] 265. The composition of embodiment 260, wherein the lipid binding protein-based complex is CER-522.

[0776] 266. The composition of embodiment 260, wherein the lipid binding protein-based complex is delipidated HDL.

[0777] 267. The composition of any one of embodiments 239 to 259, wherein the lipid binding protein-based complex is an Apomer.

[0778] 268. The composition of any one of embodiments 239 to 259, wherein the lipid binding protein-based complex is a cargomer.

[0779] 269. The composition of any one of embodiments 239 to 268, wherein the one or more ophthalmic drugs comprise a steroid, a kinase inhibitor, an angiotensin II receptor antagonist, an aldose reductase inhibitor, an immunosuppressant, a carbonic anhydrase inhibitor, an antimicrobial agent, an antiviral agent, an antihistamine, an anti-inflammatory agent, or a combination thereof.

[0780] 270. The composition of any one of embodiments 239 to 269, wherein the one or more ophthalmic drugs include azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5

[0013] In some embodiments, the present invention comprises: a) 5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol; b) 1-(2-(4-(6-nitrophenyl)pyrido[2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea; c) 1-(2-(4-(6-nitrophenyl)pyrido[2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea; d ...

[0781] 271. The composition of any one of embodiments 239 to 269, wherein the one or more ophthalmic drugs include azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)-1,4-dihydropyridine-3-carboxamide),

[0013] The invention relates to an antibiotic comprising: a monoclonal antibody comprising: a monoclonal antibody comprising: a monoclonal antibody, ...

[0782] 272. The composition of embodiment 270 or embodiment 271, wherein the one or more ophthalmic drugs comprises azithromycin, optionally wherein the concentration of azithromycin in the composition is 10 mg / ml.

[0783] 273. The composition of any one of embodiments 270 to 272, wherein the one or more ophthalmic drugs comprises spironolactone, optionally wherein the concentration of spironolactone in the composition is 1.5 mg / ml.

[0784] 274. The composition of any one of embodiments 270 to 273, wherein the one or more ophthalmic drugs comprises dexamethasone palmitate, optionally wherein the concentration of dexamethasone palmitate in the composition is 1 mg / ml.

[0785] 275. The composition of any one of embodiments 270 to 274, wherein the one or more ophthalmic drugs comprises cyclosporine, optionally wherein the concentration of cyclosporine in the composition is 1 mg / ml.

[0786] 276. The composition of any one of embodiments 239 to 275, wherein the one or more ophthalmic drugs comprises latanoprost, trovaprost, bimatoprost, tafluprost, or a combination thereof.

[0787] 277. The composition of embodiment 276, wherein the one or more ophthalmic drugs comprises latanoprost.

[0788] 278. The composition of any one of embodiments 239 to 277, wherein the one or more ophthalmic drugs comprises dexamethasone.

[0789] 279. The composition of any one of embodiments 239 to 278, wherein the one or more ophthalmic drugs comprises loteprednol etabonate.

[0790] 280. The composition of any one of embodiments 239 to 279, wherein the one or more ophthalmic drugs comprises triamcinolone acetonide.

[0791] 281. The composition of any one of embodiments 239 to 280, wherein the one or more ophthalmic drugs comprises acyclovir.

[0792] 282. The composition of any one of embodiments 239 to 281, wherein the one or more ophthalmic drugs comprises trovaprost.

[0793] 283. The composition of any one of embodiments 239 to 282, wherein the one or more ophthalmic drugs comprises bimatoprost.

[0794] 284. The composition of any one of embodiments 239 to 283, wherein the one or more ophthalmic drugs comprises tafluprost.

[0795] 285. The composition of any one of embodiments 239 to 284, wherein the one or more ophthalmic drugs comprises pazopanib.

[0796] 286. The composition of any one of embodiments 239 to 285, wherein the one or more ophthalmic drugs comprises sirolimus.

[0797] 287. The composition of any one of embodiments 239 to 286, wherein the one or more ophthalmic drugs comprises tacrolimus.

[0798] 288. The composition of any one of embodiments 239 to 287, wherein the one or more ophthalmic drugs comprises nepafenac.

[0799] 289. The composition of any one of embodiments 239 to 288, which does not comprise a cell penetrating peptide.

[0800] 290. The composition of any one of embodiments 239 to 289, which does not comprise a chemical penetration enhancer.

[0801] 291. The composition of any one of embodiments 239 to 290, which does not comprise a cytophilic peptide.

[0802] 292. The composition of any one of embodiments 239 to 291, which is a pharmaceutical composition further comprising one or more buffers, preservatives, excipients, diluents, or a combination thereof.

[0803] 293. The composition of any one of embodiments 239 to 292, for use in the method of any one of embodiments 1 to 222.

[0804] 294. A method for producing a composition comprising a lipid-binding protein-based complex and one or more ophthalmic drugs, optionally wherein the composition is the composition of any one of embodiments 251 to 293, the method comprising thermally cycling a mixture comprising the lipid-binding protein-based complex and one or more ophthalmic drugs.

[0805] 295. The method of embodiment 294, wherein the thermal cycle comprises

[0806] (a) heating the mixture from a temperature within a first temperature range to a temperature within a second temperature range,

[0807] (b) cooling the mixture of (a) from a temperature within the second temperature range to a temperature within the first temperature range; and

[0808] (c) optionally repeating steps (a) and (b) at least once.

[0809] 296. The method of embodiment 295, wherein steps (a) and (b) are repeated once.

[0810] 297. The method of embodiment 295, wherein steps (a) and (b) are repeated twice.

[0811] 298. The method of embodiment 295, wherein steps (a) and (b) are repeated three times.

[0812] 299. The method of embodiment 295, wherein steps (a) and (b) are repeated four times.

[0813] 300. The method of embodiment 295, wherein steps (a) and (b) are repeated five times.

[0814] 301. The method of any one of embodiments 295 to 300, wherein the first temperature range is 30°C to 45°C.

[0815] 302. The method of embodiment 301, wherein the temperature within the first temperature range is 37°C.

[0816] 303. The method of any one of embodiments 295 to 302, wherein the second temperature range is 50°C to 65°C.

[0817] 304. The method of embodiment 303, wherein the temperature within the second temperature range is 55°C.

[0818] 305. The method of any one of embodiments 294 to 303, comprising thermally cycling the mixture between 37°C and 55°C.

[0819] 306. A composition produced by a method comprising the method of any one of embodiments 294 to 305, optionally wherein the method further comprises the step of combining the product of the method with one or more buffers, preservatives, excipients, diluents or a combination thereof.

[0820] 307. A lipid binding protein-based complex for use in treating an eye disease in a subject, optionally in combination with one or more ophthalmic drugs, wherein the amount of the lipid binding protein-based complex administered is effective to reduce the severity of the eye disease.

[0821] 308. The lipid-binding protein-based complex used in embodiment 307, wherein the eye disease is a disease associated with lipid accumulation; preferably, the eye disease is selected from eye diseases associated with LCAT deficiency, such as fish eye disease; dry eye disease, such as dry eye disease associated with meibomian gland dysfunction (MGD) or lacrimal gland dysfunction; blepharitis; inflammatory eye diseases, such as uveitis; corneal diseases, such as lipid keratopathy; macular edema; macular degeneration; retinal detachment; eye tumors; fungal infections; viral infections; bacterial infections; multifocal choroiditis; diabetic retinopathy; proliferative vitreoretinopathy (PVR); sympathetic ophthalmia; Vogt Koyanagi-Harada (VKH) syndrome; histoplasmosis; uveal spread; vascular occlusion; and endophthalmitis.

[0822] 309. The lipid binding protein-based complex for use in any one of embodiments 307 or 308, wherein the eye disease is fish eye disease and the subject is homozygous or heterozygous for a LCAT mutation.

[0823] 310. The lipid-binding protein-based complex used in any one of embodiments 307 to 309, wherein the lipid-binding protein-based complex is a reconstituted HDL, a HDL mimetic, a cargomer or an apomer; preferably, the lipid-binding protein-based complex is selected from CER-001, CSL-111, CSL-112, CER-522 or ETC-216; more preferably, the lipid-binding protein-based complex is CER-001.

[0824] 311. The lipid-binding protein-based complex used in any one of embodiments 307 to 310, wherein the lipid-binding protein-based complex comprises one or more ophthalmic drugs complexed with the lipid-binding protein-based complex, and wherein the one or more ophthalmic drugs include steroids, kinase inhibitors, angiotensin II receptor antagonists, aldose reductase inhibitors, immunosuppressants, carbonic anhydrase inhibitors, antimicrobial agents, antiviral agents, antihistamines, anti-inflammatory agents or combinations thereof.

[0825] 312. The lipid binding protein-based complex used in any one of embodiments 307 to 311, wherein the one or more ophthalmic drugs include azithromycin, spironolactone, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl) Pyrido[2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, 2-methylsorbitol, sirolimus, Acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, latanoprost, trovaprost, bimatoprost, tafluprost, dexamethasone palmitate, cyclosporine, or a combination thereof.

[0826] 313. The lipid-binding protein-based complex used in any one of embodiments 307 to 312, wherein the lipid-binding protein-based complex is administered according to a dosing regimen comprising:

[0827] (a) induction regimen; and / or

[0828] (b) a consolidation programme; and / or

[0829] (c) Maintenance Program.

[0830] 314. The lipid-binding protein-based complex used in embodiment 313, wherein the dosing regimen comprises an induction regimen comprising administering multiple doses of the lipid-binding protein-based complex to the subject, wherein the multiple doses are separated by 1 day or more.

[0831] 315. The lipid-binding protein-based complex used in embodiment 314, wherein the dosage interval after the initial dose of the induction regimen is no more than 3 days, preferably the dosage interval after the initial dose of the induction regimen is 1 to 3 days.

[0832] 316. The lipid-binding protein-based complex used in any one of embodiments 313 to 315, wherein the induction regimen lasts for at least one week, preferably the induction regimen lasts for two weeks, more preferably lasts for three weeks.

[0833] 317. The lipid-binding protein-based complex used in any one of embodiments 313 to 316, wherein the induction regimen comprises administering three doses of the lipid-binding protein-based complex to the subject per week.

[0834] 318. The lipid-binding protein-based complex used in any one of embodiments 313 to 317, wherein the induction regimen comprises administering to the subject at least three doses of the lipid-binding protein-based complex; preferably four doses or more, five doses or more, six doses or more, seven doses or more, eight doses or more or nine doses or more; more preferably, the induction regimen comprises administering to the subject nine doses or more of the lipid-binding protein-based complex.

[0835] 319. The lipid-binding protein-based complex used in any one of embodiments 313 to 318, wherein the induction regimen comprises administering a first dose of the lipid-binding protein-based complex to the subject on day 1 and administering subsequent doses of the induction regimen to the subject on days 2, 4, 7, 9, 11, 14, 16 and 18.

[0836] 320. The lipid-binding protein-based complex used in any one of embodiments 313 to 319, wherein the dosing regimen comprises a consolidation regimen comprising administering multiple doses of the lipid-binding protein-based complex to the subject, wherein the multiple doses are separated by 2 or more days.

[0837] 321. The lipid binding protein-based complex used in any one of embodiments 313 to 320, wherein the doses of the consolidation regimen are separated by no more than four days, preferably the doses of the consolidation regimen are separated by three or four days from each other.

[0838] 322. The lipid-binding protein-based complex used in any one of embodiments 313 to 321, wherein the consolidation regimen lasts for at least three weeks.

[0839] 323. The lipid-binding protein-based complex used in any one of embodiments 313 to 322, wherein the consolidation regimen comprises administering at least two doses of the lipid-binding protein-based complex to the subject per week.

[0840] 324. The lipid-binding protein-based complex used in any one of embodiments 313 to 323, wherein the consolidation regimen comprises administering to the subject at least two doses of the lipid-binding protein-based complex; preferably three doses or more, four doses or more, five doses or more, or six doses or more; more preferably, the consolidation regimen comprises administering to the subject six doses or more of the lipid-binding protein-based complex.

[0841] 325. The lipid-binding protein-based complex used in any one of embodiments 313 to 324, wherein the consolidation regimen comprises administering six doses of the lipid-binding protein-based complex to the subject on days 21, 24, 28, 31, 35 and 38 following the induction regimen starting on day 1.

[0842] 326. The lipid-binding protein-based complex used in any one of embodiments 313 to 325, wherein the dosing regimen comprises a maintenance regimen comprising administering a dose of the lipid-binding protein-based complex to the subject every 3 days or more, preferably once every 5 days or more, and more preferably once a week.

[0843] 327. The lipid-binding protein-based complex used in any one of embodiments 313 to 326, wherein the maintenance regimen comprises administering the lipid-binding protein-based complex to the subject for at least one month.

[0844] 328. The lipid-binding protein-based complex used in any one of embodiments 313 to 327, wherein the dose of the lipid-binding protein-based complex administered in the induction regimen, consolidation regimen and / or maintenance regimen is 4 to 30 mg / kg (based on protein weight); preferably 5 to 15 mg / kg (based on protein weight), 10 to 20 mg / kg (based on protein weight) or 15 to 25 mg / kg (based on protein weight).

[0845] 329. The lipid-binding protein-based complex used in any one of embodiments 313 to 328, wherein the dose of the lipid-binding protein-based complex administered in the induction regimen, consolidation regimen and / or maintenance regimen is 8 mg / kg (based on protein weight) or 10 mg / kg (based on protein weight).

[0846] 330. The lipid-binding protein-based complex used in any one of embodiments 313 to 329, wherein the dose of the lipid-binding protein-based complex administered in the induction regimen, consolidation regimen and / or maintenance regimen is 300 mg to 3000 mg; preferably 300 mg to 1500 mg, 400 mg to 1500 mg, 500 mg to 1200 mg or 500 mg to 1000 mg.

[0847] 331. The lipid-binding protein-based complex for use in any one of embodiments 307 to 330, wherein the lipid-binding protein-based complex is administered peripherally, optionally by infusion.

[0848] 332. The lipid-binding protein-based complex used in any one of embodiments 307 to 330, wherein the lipid-binding protein-based complex is administered topically, such as intraocularly, such as by intraocular injection (e.g., suprachoroidal injection or suprascleral injection), or topically, such as by eye drops or via an implant (e.g., a biodegradable or non-biodegradable disc, sheet, plug, rod or pellet).

[0849] 333. The lipid binding protein-based complex for use in any one of embodiments 207 to 332, wherein an antihistamine is administered prior to administration of one or more doses of the lipid binding protein-based complex.

[0850] 334. A lipid-binding protein-based complex used in any one of embodiments 307 to 333, wherein the subject is also treated with a lipid-controlling drug; preferably, the lipid-controlling drug comprises a statin, such as atorvastatin, rosuvastatin, simvastatin, fluvastatin, lovastatin or pravastatin; a cholesterol absorption inhibitor, such as ezetimibe; niacin; aspirin; a proprotein convertase subtilisin / kexin type 9 (PCSK9) inhibitor, such as an antibody selected from alirocumab, bococizumab, evolocumab, 1D05-IgG2 or LY3015014, or an RNAi therapeutic agent, such as ALN-PCSSC.

[0851] 335. The lipid binding protein-based complex for use in any one of embodiments 307 to 334, wherein the subject is also treated with a standard of care therapy for an eye disease.

[0852] 336. The lipid-binding protein-based complex for use in any one of embodiments 307 to 335, wherein the lipid-binding protein-based complex does not comprise a cell penetrating peptide and is not administered together with a cell penetrating peptide.

[0853] 337. The lipid-binding protein-based complex for use in any one of embodiments 307 to 336, wherein the lipid-binding protein-based complex does not contain a chemical permeation enhancer and is not administered together with a chemical permeation enhancer.

[0854] 338. The lipid-binding protein-based complex used in any one of embodiments 307 to 337, wherein the lipid-binding protein-based complex does not comprise a cytophilic peptide and is not administered together with a cytophilic peptide.

[0855] 339. A lipoprotein complex for use in treating an eye disease in a subject in need thereof.

[0856] wherein the lipoprotein complex comprises an ApoA-I apolipoprotein portion and a lipid portion comprising one or more phospholipids, and

[0857] wherein the subject has ocular lipid deposition.

[0858] 340. The lipoprotein complex for use according to embodiment 339, wherein the lipoprotein complex comprises an ApoA-I apolipoprotein portion and a lipid portion, wherein the lipid portion comprises at least one neutral phospholipid and, optionally, one or more negatively charged phospholipids.

[0859] 341. The lipoprotein complex for use according to embodiment 339 or embodiment 340, wherein the lipoprotein complex is selected from CER-001, CSL-111 and ETC-216.

[0860] 342. The lipoprotein complex for use according to any one of embodiments 339 to 341, wherein the lipoprotein complex comprises an ApoA-I apolipoprotein portion and a lipoprotein portion comprising sphingomyelin and one or more negatively charged phospholipids.

[0861] 343. A lipoprotein complex for use according to any one of embodiments 339 to 342, wherein the lipoprotein complex comprises an ApoA-I apolipoprotein portion and a lipid portion, wherein the lipid portion essentially consists of sphingomyelin and about 0.2 to 6 wt % of negatively charged phospholipids, and wherein the molar ratio of the lipid portion to the ApoA-I apolipoprotein portion ranges from about 2:1 to 200:1.

[0862] 344. The lipoprotein complex for use according to any one of embodiments 340 to 343, wherein the negatively charged phospholipid comprises 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DPPG) or a salt thereof.

[0863] 345. The lipoprotein complex for use according to any one of embodiments 339 to 344, wherein the lipoprotein complex is CER-001.

[0864] 346. The lipoprotein complex for use according to any one of embodiments 339 to 345, wherein the ocular lipid deposition is corneal lipid deposition, retinal lipid deposition, eyelid lipid deposition or a combination thereof.

[0865] 347. The lipoprotein complex used in any one of embodiments 339 to 346, wherein the eye disease is selected from dry eye associated with lipid accumulation, including dry eye associated with meibomian gland dysfunction (MGD) and dry eye associated with tear gland dysfunction, blepharitis, inflammatory eye diseases, uveitis including anterior uveitis, intermediate uveitis, posterior uveitis and panuveitis, corneal diseases including lipid keratopathy, eye diseases associated with LCAT deficiency including fish eye disease, dry macular degeneration (dry AMD), Stargardt disease and Leber idiopathic stellate neuroretinitis, macular edema, macular degeneration, retinal detachment, eye tumors, fungal infections, viral infections, bacterial infections including bacterial conjunctivitis and trachoma, multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis and glaucoma.

[0866] 348. The lipoprotein complex for use according to any one of embodiments 339 to 347, wherein the lipoprotein complex is administered in an amount effective to reduce ocular lipid deposition in a subject.

[0867] 349. The lipoprotein complex for use according to any one of embodiments 339 to 348, wherein the lipoprotein complex further comprises one or more ophthalmic drugs.

[0868] 350. The lipoprotein complex for use according to embodiment 349, wherein the ophthalmic drug is a steroid, a kinase inhibitor, an angiotensin II receptor antagonist, an aldose reductase inhibitor, an immunosuppressant, a carbonic anhydrase inhibitor, an antimicrobial agent, an antiviral agent, an antihistamine, an anti-inflammatory agent, a prostaglandin analog or a combination thereof; preferably, the ophthalmic drug is selected from azithromycin, dexamethasone, dexamethasone palmitate, difluprednate, estradiol, fluocinolone, Fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan , olmesartan, telmisartan, valsartan, 2-methylsorbitol, sirolimus, cyclosporine, tacrolimus, acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, latanoprost, trovaprost, bimatoprost, or a combination thereof.

[0869] 351. The lipoprotein complex for use according to any one of embodiments 339 to 350, wherein the lipoprotein complex is administered peripherally, optionally by infusion.

[0870] 352. The lipoprotein complex for use according to any one of embodiments 339 to 350, wherein the lipoprotein complex is administered intraocularly, preferably by intraocular injection, more preferably by intravitreal injection, suprachoroidal injection or suprascleral injection.

[0871] 353. The lipoprotein complex for use according to any one of embodiments 339 to 350, wherein the lipoprotein complex is administered by a topical route, preferably using eye drops.

[0872] 354. The lipoprotein complex for use according to any one of embodiments 339 to 353, wherein the lipoprotein complex does not comprise a cell penetrating peptide and is not administered together with a cell penetrating peptide.

[0873] 355. The lipoprotein complex for use according to any one of embodiments 339 to 354, wherein the lipoprotein complex does not comprise a chemical permeation enhancer and is not administered together with a chemical permeation enhancer.

[0874] 356. The lipoprotein complex for use according to any one of embodiments 339 to 355, wherein the lipoprotein complex does not comprise a cytophilic peptide and is not administered together with a cytophilic peptide.

[0875] 357. A method of treating an eye disease in a subject, the method comprising:

[0876] (a) administering to the subject an apolipoprotein AI ("ApoA-I") preparation comprising ApoA-I and one or more lipids, wherein the ApoA-I and the lipids are present in the form of a lipoprotein complex, wherein:

[0877] (i) the subject suffers from cataract, uveitis, eye disease associated with lipid accumulation, lipid keratopathy, corneal dystrophy, corneal opacity, dry eye disease, blepharitis, inflammatory eye disease, macular edema, macular degeneration, retinal detachment, eye tumor, fungal infection, viral infection, bacterial infection (e.g., bacterial conjunctivitis or trachoma), multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis, glaucoma, Stargardt's disease, ocular lipid deposition; and / or

[0878] (ii) administering the formulation by suprachoroidal injection, suprascleral injection, implant, iontophoresis or electroporation.

[0879] 358. The method of embodiment 357, wherein the ApoA-I has an amino acid sequence of amino acids 25-267 of SEQ ID NO:2.

[0880] 359. The method of embodiment 357 or embodiment 358, wherein the ApoA-I is recombinantly expressed.

[0881] 360. The method of any one of embodiments 357 to 359, wherein the one or more lipids comprise neutral lipids.

[0882] 361. The method of embodiment 360, wherein the neutral lipid comprises sphingomyelin.

[0883] 362. The method of embodiment 361, wherein the neutral lipid consists of sphingomyelin.

[0884] 363. The method of embodiment 361 or embodiment 362, wherein the sphingomyelin comprises a native sphingomyelin.

[0885] 364. The method of embodiment 363, wherein the natural sphingomyelin is hen egg sphingomyelin.

[0886] 365. The method of embodiment 361 or embodiment 362, wherein the sphingomyelin comprises a synthetic sphingomyelin.

[0887] 366. The method of embodiment 365, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin.

[0888] 367. The method of any one of embodiments 357 to 362, wherein the one or more lipids further comprise a negatively charged lipid.

[0889] 368. The method of embodiment 367, wherein the negatively charged lipid comprises 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof.

[0890] 369. The method of embodiment 368, wherein the negatively charged lipid consists of 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof.

[0891] 370. The method of any one of embodiments 363 to 369, wherein the molar ratio of the negatively charged lipid to the neutral lipid to the component of ApoA-I in the formulation is 2-6:90-120:1.

[0892] 371. The method of any one of embodiments 360 to 370, wherein the lipids consist of 95 to 99% by weight of neutral phospholipids and 1 to 5% by weight of negatively charged phospholipids.

[0893] 372. The method of embodiment 371, wherein the lipids consist of 96 to 98 weight % neutral phospholipids and 2 to 4 weight % negatively charged phospholipids.

[0894] 373. The method of embodiment 372, wherein the lipids consist of 97% by weight of neutral phospholipids and 3% by weight of negatively charged phospholipids.

[0895] 374. The method of any one of embodiments 357 to 373, having a weight ratio of ApoA-I to lipid ranging from 1:2 to 1:3.

[0896] 375. The method of embodiment 374, which has a weight ratio of ApoA-I to lipid of about 1:2.7.

[0897] 376. The method of any one of embodiments 357 to 375, wherein the lipoprotein complex is at least 95% homogeneous as reflected by a single peak in gel permeation chromatography.

[0898] 377. The method of any one of embodiments 357 to 376, wherein the formulation is a carrier for or has been complexed with one or more ophthalmic drugs, optionally wherein one or more of the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

[0899] 378. The method of embodiment 377, wherein the one or more ophthalmic drugs include steroids, kinase inhibitors, angiotensin II receptor antagonists, aldose reductase inhibitors, immunosuppressants, carbonic anhydrase inhibitors, antimicrobial agents, antiviral agents, antihistamines, anti-inflammatory agents, prostaglandin analogs, or combinations thereof.

[0900] 379. The method of embodiment 377 or embodiment 378, wherein the one or more ophthalmic drugs include azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyrido[2, 3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, 2-methylsorbitol, sirolimus, cyclosporine bacteriocin, tacrolimus, acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, latanoprost, trovaprost, bimatoprost, dexamethasone palmitate, or a combination thereof.

[0901] 380. The method of any one of embodiments 357 to 379, wherein the eye disease is fish eye disease.

[0902] 381. The method of any one of embodiments 357 to 379, wherein the eye disease is secondary lipid keratopathy, a hereditary corneal dystrophy, an anterior or superficial corneal dystrophy, a stromal corneal dystrophy, or a posterior corneal dystrophy.

[0903] 382. The method of any one of embodiments 357 to 379, wherein the subject has corneal opacity and the method comprises administering an amount of the lipid binding protein-based complex effective to reduce corneal opacity in the subject.

[0904] 383. The method of any one of embodiments 357 to 379, wherein the eye disease is dry eye disease, which is (a) associated with meibomian gland dysfunction (MGD), optionally wherein the MGD is obstructive MGD or (b) associated with tear gland dysfunction.

[0905] 384. The method of any one of embodiments 357 to 379, wherein the eye disease is blepharitis, inflammatory eye disease, anterior uveitis, intermediate uveitis, posterior uveitis, or panuveitis.

[0906] 385. The method of embodiment 384, wherein the eye disease is acute anterior uveitis.

[0907] 386. The method of any one of embodiments 357 to 379, wherein the eye disease is dry macular degeneration.

[0908] 387. The method of any one of embodiments 357 to 379, wherein the eye disease is wet macular degeneration.

[0909] 388. The method of any one of embodiments 357 to 379, wherein the eye disease is diabetic macular edema.

[0910] 389. The method of any one of embodiments 357 to 388, wherein the subject has impaired vision due to an eye disease, and the method comprises administering to the subject an amount of the lipid binding protein-based complex that improves the subject's vision.

[0911] 390. The method of any one of embodiments 357 to 389, wherein the subject has ocular lipid deposition, optionally wherein the ocular lipid deposition comprises corneal lipid deposition, retinal lipid deposition, eyelid lipid deposition, or a combination thereof.

[0912] 391. An apolipoprotein AI ("ApoA-I") formulation comprising ApoA-I and one or more lipids, wherein the ApoA-I and the lipids are present in the form of a lipoprotein complex, for use in treating an eye disease in a subject, wherein:

[0913] (i) the subject suffers from cataract, uveitis, eye disease associated with lipid accumulation, lipid keratopathy, corneal dystrophy, corneal opacity, dry eye disease, blepharitis, inflammatory eye disease, macular edema (e.g., diabetic macular edema), macular degeneration, retinal detachment, eye tumor, fungal infection, viral infection, bacterial infection (e.g., bacterial conjunctivitis or trachoma), multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis, glaucoma, Stargardt's disease, ocular lipid deposition; and / or

[0914] (ii) administering the formulation by suprachoroidal injection, suprascleral injection, implant, iontophoresis or electroporation.

[0915] 392. A formulation for use according to embodiment 391, wherein the ApoA-I has an amino acid sequence of amino acids 25-267 of SEQ ID NO:2.

[0916] 393. A formulation for use according to embodiment 391 or embodiment 392, wherein the ApoA-I is recombinantly expressed.

[0917] 394. The formulation of any one of embodiments 391 to 393, wherein the one or more lipids comprise a neutral lipid.

[0918] 395. The formulation of embodiment 394, wherein the neutral lipid comprises sphingomyelin.

[0919] 396. The formulation of embodiment 395, wherein the neutral lipid consists of sphingomyelin.

[0920] 397. A formulation for use according to embodiment 395 or embodiment 396, wherein the sphingomyelin comprises native sphingomyelin.

[0921] 398. The formulation for use according to embodiment 397, wherein the natural sphingomyelin is hen egg sphingomyelin.

[0922] 399. A formulation for use according to embodiment 395 or embodiment 396, wherein the sphingomyelin comprises a synthetic sphingomyelin.

[0923] 400. The formulation for use according to embodiment 399, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin.

[0924] 401. A formulation for use according to any one of embodiments 391 to 396, wherein the one or more lipids further comprise negatively charged lipids.

[0925] 402. A formulation for use according to embodiment 401, wherein the negatively charged lipid comprises 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof.

[0926] 403. A formulation for use according to embodiment 402, wherein the negatively charged lipid consists of 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof.

[0927] 404. A formulation for use according to any one of embodiments 397 to 403, wherein the molar ratio of negatively charged lipids to neutral lipids to components of ApoA-I in the formulation is 2-6:90-120:1.

[0928] 405. The formulation for use according to any one of embodiments 394 to 404, wherein the lipids consist of 95 to 99% by weight of neutral phospholipids and 1 to 5% by weight of negatively charged phospholipids.

[0929] 406. The method of embodiment 405, wherein the lipids consist of 96 to 98 weight % neutral phospholipids and 2 to 4 weight % negatively charged phospholipids.

[0930] 407. The formulation for use according to embodiment 406, wherein the lipids consist of 97% by weight of neutral phospholipids and 3% by weight of negatively charged phospholipids.

[0931] 408. A formulation for use according to any one of embodiments 391 to 407, having a weight ratio of ApoA-I to lipid in the range of 1:2 to 1:3.

[0932] 409. A formulation for use according to embodiment 408, which has a weight ratio of ApoA-I to lipid of about 1:2.7.

[0933] 410. The formulation for use according to any one of embodiments 391 to 409, wherein the lipoprotein complex is at least 95% homogeneous as reflected by a single peak in gel permeation chromatography.

[0934] 411. A formulation for use according to any one of embodiments 391 to 410, wherein the formulation is a carrier for or has been complexed with one or more ophthalmic drugs, optionally wherein one or more of the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

[0935] 412. The formulation for use in embodiment 411, wherein the one or more ophthalmic drugs include steroids, kinase inhibitors, angiotensin II receptor antagonists, aldose reductase inhibitors, immunosuppressants, carbonic anhydrase inhibitors, antimicrobial agents, antiviral agents, antihistamines, anti-inflammatory agents, prostaglandin analogs, or combinations thereof.

[0936] 413. A formulation for use according to embodiment 411 or embodiment 412, wherein the one or more ophthalmic drugs include azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyridinol[ 2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, 2-methylsorbitol, sirolimus, cyclopentadiene The invention relates to a pharmaceutical composition comprising the following: acyclovir, chloramphenicol, chlortetracycline, tacrolimus, acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, latanoprost, trovaprost, bimatoprost, dexamethasone palmitate, or a combination thereof.

[0937] 414. The formulation for use according to any one of embodiments 391 to 413, wherein the eye disease is fish eye disease.

[0938] 415. The formulation for use according to any one of embodiments 391 to 413, wherein the eye disease is secondary lipid keratopathy, a hereditary corneal dystrophy, anterior or superficial corneal dystrophy, stromal corneal dystrophy or posterior corneal dystrophy.

[0939] 416. A formulation for use according to any one of embodiments 391 to 413, wherein the subject suffers from corneal opacity and the method comprises administering an amount of the lipid binding protein-based complex effective to reduce corneal opacity in the subject.

[0940] 417. A formulation for use according to any one of embodiments 391 to 413, wherein the eye disease is dry eye disease, which is (a) associated with meibomian gland dysfunction (MGD), optionally wherein the MGD is obstructive MGD or (b) associated with tear gland dysfunction.

[0941] 418. The formulation for use according to any one of embodiments 391 to 413, wherein the eye disease is blepharitis, inflammatory eye disease, anterior uveitis, intermediate uveitis, posterior uveitis or panuveitis.

[0942] 419. The formulation for use according to embodiment 418, wherein the eye disease is acute anterior uveitis.

[0943] 420. A formulation for use according to any one of embodiments 391 to 413, wherein the eye disease is dry macular degeneration.

[0944] 421. A formulation for use according to any one of embodiments 391 to 413, wherein the eye disease is wet macular degeneration.

[0945] 422. The formulation for use according to any one of embodiments 391 to 413, wherein the eye disease is diabetic macular edema.

[0946] 423. A formulation for use according to any one of embodiments 391 to 422, wherein the subject has impaired vision due to an eye disease, and the method comprises administering to the subject an amount of the lipid binding protein-based complex that improves the subject's vision.

[0947] 424. A formulation for use according to any one of embodiments 391 to 423, wherein the subject has ocular lipid deposition, optionally wherein the ocular lipid deposition comprises corneal lipid deposition, retinal lipid deposition, eyelid lipid deposition or a combination thereof.

[0948] 425. An apolipoprotein AI ("ApoA-I") preparation comprising ApoA-I and one or more lipids, wherein the ApoA-I and the lipids are present in the form of a lipoprotein complex, for use in the preparation of a medicament for treating an eye disease in a subject, wherein:

[0949] (i) the subject suffers from cataract, uveitis, eye disease associated with lipid accumulation, lipid keratopathy, corneal dystrophy, corneal opacity, dry eye disease, blepharitis, inflammatory eye disease, macular edema (e.g., diabetic macular edema), macular degeneration, retinal detachment, eye tumor, fungal infection, viral infection, bacterial infection (e.g., bacterial conjunctivitis or trachoma), multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis, glaucoma, Stargardt's disease, ocular lipid deposition; and / or

[0950] (ii) administering the formulation by suprachoroidal injection, suprascleral injection, implant, iontophoresis or electroporation.

[0951] 426. A formulation for use according to embodiment 425, wherein the ApoA-I has an amino acid sequence of amino acids 25-267 of SEQ ID NO:2.

[0952] 427. A formulation for use according to embodiment 425 or embodiment 426, wherein the ApoA-I is recombinantly expressed.

[0953] 428. The formulation of any one of embodiments 425 to 427, wherein the one or more lipids comprise a neutral lipid.

[0954] 429. The formulation of embodiment 428, wherein the neutral lipid comprises sphingomyelin.

[0955] 430. The formulation of embodiment 429, wherein the neutral lipid consists of sphingomyelin.

[0956] 431. A formulation for use according to embodiment 429 or embodiment 430, wherein the sphingomyelin comprises native sphingomyelin.

[0957] 432. A formulation for use according to embodiment 431, wherein the natural sphingomyelin is hen egg sphingomyelin.

[0958] 433. A formulation for use according to embodiment 429 or embodiment 430, wherein the sphingomyelin comprises a synthetic sphingomyelin.

[0959] 434. A formulation for use according to embodiment 433, wherein the synthetic sphingomyelin is palmitoyl sphingomyelin.

[0960] 435. A formulation for use according to any one of embodiments 425 to 430, wherein the one or more lipids further comprise a negatively charged lipid.

[0961] 436. A formulation for use according to embodiment 435, wherein the negatively charged lipid comprises 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof.

[0962] 437. A formulation for use according to embodiment 436, wherein the negatively charged lipid consists of 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] ("DPPG") or a salt thereof.

[0963] 438. A formulation for use according to any one of embodiments 431 to 437, wherein the molar ratio of negatively charged lipids to neutral lipids to components of ApoA-I in the formulation is 2-6:90-120:1.

[0964] 439. A formulation for use according to any one of embodiments 428 to 438, wherein the lipids consist of 95 to 99% by weight of neutral phospholipids and 1 to 5% by weight of negatively charged phospholipids.

[0965] 440. The method of embodiment 439, wherein the lipids consist of 96 to 98 weight % neutral phospholipids and 2 to 4 weight % negatively charged phospholipids.

[0966] 441. A formulation for use according to embodiment 440, wherein the lipids consist of 97% by weight of neutral phospholipids and 3% by weight of negatively charged phospholipids.

[0967] 442. A formulation for use according to any one of embodiments 425 to 441, having a weight ratio of ApoA-I to lipid in the range of 1:2 to 1:3.

[0968] 443. A formulation for use according to embodiment 442, which has a weight ratio of ApoA-I to lipid of about 1:2.7.

[0969] 444. The formulation for use according to any one of embodiments 425 to 443, wherein the lipoprotein complex is at least 95% homogeneous as reflected by a single peak in gel permeation chromatography.

[0970] 445. A formulation for use according to any one of embodiments 425 to 444, wherein the formulation is a carrier for or has been complexed with one or more ophthalmic drugs, optionally wherein one or more of the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

[0971] 446. The formulation for use in embodiment 445, wherein the one or more ophthalmic drugs comprise a steroid, a kinase inhibitor, an angiotensin II receptor antagonist, an aldose reductase inhibitor, an immunosuppressant, a carbonic anhydrase inhibitor, an antimicrobial agent, an antiviral agent, an antihistamine, an anti-inflammatory agent, a prostaglandin analog, or a combination thereof.

[0972] 447. The formulation for use according to embodiment 445 or embodiment 446, wherein the one or more ophthalmic drugs include azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxyphenyl)pyridinol[ 2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, 2-methylsorbitol, sirolimus, cyclopentadiene The invention relates to a pharmaceutical composition comprising the following: acyclovir, chloramphenicol, chlortetracycline, tacrolimus, acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, latanoprost, trovaprost, bimatoprost, dexamethasone palmitate, or a combination thereof.

[0973] 448. The formulation for use according to any one of embodiments 425 to 447, wherein the eye disease is fish eye disease.

[0974] 449. The formulation for use according to any one of embodiments 425 to 447, wherein the eye disease is secondary lipid keratopathy, a hereditary corneal dystrophy, anterior or superficial corneal dystrophy, stromal corneal dystrophy or posterior corneal dystrophy.

[0975] 450. A formulation for use according to any one of embodiments 425 to 447, wherein the subject suffers from corneal opacity and the method comprises administering an amount of the lipid binding protein-based complex effective to reduce corneal opacity in the subject.

[0976] 451. A formulation for use according to any one of embodiments 425 to 447, wherein the eye disease is dry eye disease, which is (a) associated with meibomian gland dysfunction (MGD), optionally wherein the MGD is obstructive MGD or (b) associated with tear gland dysfunction.

[0977] 452. A formulation for use according to any one of embodiments 425 to 447, wherein the eye disease is blepharitis, inflammatory eye disease, anterior uveitis, intermediate uveitis, posterior uveitis or panuveitis.

[0978] 453. A formulation for use according to embodiment 452, wherein the eye disease is acute anterior uveitis.

[0979] 454. A formulation for use according to any one of embodiments 425 to 447, wherein the eye disease is dry macular degeneration.

[0980] 455. A formulation for use according to any one of embodiments 425 to 447, wherein the eye disease is wet macular degeneration.

[0981] 456. A formulation for use according to any one of embodiments 425 to 447, wherein the eye disease is diabetic macular edema.

[0982] 457. A formulation for use according to any one of embodiments 425 to 456, wherein the subject has impaired vision due to an eye disease, and the method comprises administering to the subject an amount of the lipid binding protein-based complex that improves the subject's vision.

[0983] 458. A formulation for use according to any one of embodiments 425 to 457, wherein the subject suffers from ocular lipid deposition, optionally wherein the ocular lipid deposition comprises corneal lipid deposition, retinal lipid deposition, eyelid lipid deposition or a combination thereof.

[0984] Various aspects of the present disclosure are described in the embodiments set forth in the following numbered paragraphs of Group 2.

[0985] Group 2:

[0986] 1. A lipid binding protein-based complex for use in a method for treating an eye disease in a subject, wherein the lipid binding protein-based complex (a) is CER-001 and / or (b) is a carrier for one or more ophthalmic drugs, optionally wherein:

[0987] (a) the subject suffers from cataract or acute anterior uveitis; and / or

[0988] (b) The lipid binding protein-based complex is administered by suprachoroidal injection, suprascleral injection, implant, iontophoresis or electroporation.

[0989] 2. The lipid-binding protein-based complex for use according to embodiment 1, wherein the eye disease is a disease associated with lipid accumulation.

[0990] 3. The lipid binding protein-based complex for use according to embodiment 2, wherein the eye disease is fish eye disease.

[0991] 4. The lipid-binding protein-based complex for use according to embodiment 2, wherein the eye disease is lipid keratopathy, optionally wherein the lipid keratopathy is secondary lipid keratopathy.

[0992] 5. The lipid binding protein-based complex for use according to embodiment 2, wherein the eye disease is a corneal dystrophy, such as a hereditary corneal dystrophy, anterior or superficial corneal dystrophy, stromal corneal dystrophy or posterior corneal dystrophy.

[0993] 6. The lipid-binding protein-based complex for use according to any one of embodiments 1 to 5, wherein the subject suffers from corneal opacity, and the method comprises administering the lipid-binding protein-based complex in an amount effective to reduce corneal opacity in the subject.

[0994] 7. The lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the subject suffers from cataracts.

[0995] 8. A lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is dry eye, optionally wherein the dry eye disease is (a) associated with meibomian gland dysfunction (MGD), optionally wherein the MGD is obstructive MGD or (b) associated with lacrimal gland dysfunction.

[0996] 9. The lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is blepharitis.

[0997] 10. The lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is an inflammatory eye disease.

[0998] 11. A lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is uveitis, optionally wherein the uveitis is anterior uveitis, intermediate uveitis, posterior uveitis or panuveitis.

[0999] 12. The lipid binding protein-based complex for use according to embodiment 11, wherein the eye disease is acute anterior uveitis.

[1000] 13. The lipid-binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is macular edema, macular degeneration, retinal detachment, eye tumor, fungal infection, viral infection, bacterial infection (e.g., bacterial conjunctivitis or trachoma), multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis or glaucoma.

[1001] 14. The lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is dry-type macular degeneration.

[1002] 15. The lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is wet macular degeneration.

[1003] 16. The lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is Stargardt's disease.

[1004] 17. The lipid binding protein-based complex for use according to embodiment 1 or embodiment 2, wherein the eye disease is diabetic retinopathy, optionally wherein the subject suffers from diabetic macular edema.

[1005] 18. The lipid-binding protein-based complex for use according to any one of embodiments 1 to 17, wherein the subject has impaired vision due to an eye disease, and the method comprises administering to the subject an amount of the lipid-binding protein-based complex that improves the subject's vision.

[1006] 19. The lipid-binding protein-based complex for use according to any one of embodiments 1 to 18, wherein the subject suffers from ocular lipid deposition, optionally wherein the ocular lipid deposition comprises corneal lipid deposition, retinal lipid deposition, eyelid lipid deposition or a combination thereof.

[1007] 20. The lipid binding protein based complex for use according to any one of embodiments 1 to 19, wherein the lipid binding protein based complex is CER-001.

[1008] 21. A lipid-binding protein-based complex for use according to any one of embodiments 1 to 20, wherein the lipid-binding protein-based complex is a carrier of one or more ophthalmic drugs, optionally wherein (I) the lipid-binding protein-based complex is CER-001, CSL-111, CSL-112, ETC-216, CER-522, delipidated HDL, Apomer or Cargomer and / or (II) the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

[1009] 22. A lipid binding protein-based complex for use according to embodiment 21, wherein the one or more ophthalmic drugs include steroids, kinase inhibitors, angiotensin II receptor antagonists, aldose reductase inhibitors, immunosuppressants, carbonic anhydrase inhibitors, antimicrobial agents, antiviral agents, antihistamines, anti-inflammatory agents, prostaglandin analogs or combinations thereof.

[1010] 23. The lipid binding protein-based complex for use according to embodiment 21 or embodiment 22, wherein the one or more ophthalmic drugs include azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5-dimethoxy phenyl)pyrido[2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, val...

Claims

1. A lipid binding protein-based complex for use in a method of treating an eye disease in a subject, wherein the lipid binding protein-based complex (a) is CER-001 and / or (b) is a carrier of one or more ophthalmic drugs, wherein: (a) the subject suffers from cataract or acute anterior uveitis; and / or (b) The lipid binding protein-based complex is administered by suprachoroidal injection, suprascleral injection, implant, iontophoresis or electroporation.

2. The lipid binding protein-based complex for use according to claim 1, wherein the eye disease is a disease associated with lipid accumulation.

3. The lipid binding protein based complex for use according to claim 2, wherein the eye disease is fish eye disease.

4. The lipid binding protein based complex for use according to claim 2, wherein the eye disease is lipid keratopathy, optionally wherein the lipid keratopathy is secondary lipid keratopathy.

5. The lipid binding protein based complex for use according to claim 2, wherein the eye disease is a corneal dystrophy, such as a hereditary corneal dystrophy, anterior or superficial corneal dystrophy, stromal corneal dystrophy or posterior corneal dystrophy.

6. The lipid-binding protein-based complex for use according to any one of claims 1 to 5, wherein the subject suffers from corneal opacity and the method comprises administering an amount of the lipid-binding protein-based complex effective to reduce corneal opacity in the subject.

7. The lipid binding protein based complex for use according to claim 1 or claim 2, wherein the subject suffers from cataracts.

8. A lipid binding protein based complex for use according to claim 1 or claim 2, wherein the eye disease is dry eye, optionally wherein the dry eye disease is (a) associated with meibomian gland dysfunction (MGD), optionally wherein the MGD is obstructive MGD or (b) associated with lacrimal gland dysfunction.

9. The lipid binding protein based complex for use according to claim 1 or claim 2, wherein the eye disease is blepharitis.

10. The lipid binding protein based complex for use according to claim 1 or claim 2, wherein the eye disease is an inflammatory eye disease.

11. A lipid binding protein based complex for use according to claim 1 or claim 2, wherein the eye disease is uveitis, optionally wherein the uveitis is anterior uveitis, intermediate uveitis, posterior uveitis or panuveitis.

12. The lipid binding protein based complex for use according to claim 11, wherein the eye disease is acute anterior uveitis.

13. The lipid binding protein-based complex for use according to claim 1 or claim 2, wherein the eye disease is macular edema, macular degeneration, retinal detachment, eye tumor, fungal infection, viral infection, bacterial infection (e.g., bacterial conjunctivitis or trachoma), multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis or glaucoma.

14. A lipid binding protein based complex for use according to claim 1 or claim 2, wherein the eye disease is dry type macular degeneration.

15. A lipid binding protein based complex for use according to claim 1 or claim 2, wherein the eye disease is wet macular degeneration.

16. A lipid binding protein based complex for use according to claim 1 or claim 2, wherein the eye disease is Stargardt's disease.

17. A lipid binding protein based complex for use according to claim 1 or claim 2, wherein the eye disease is diabetic retinopathy, optionally wherein the subject suffers from diabetic macular edema.

18. The lipid binding protein-based complex for use according to any one of claims 1 to 17, wherein the subject has impaired vision due to an eye disease, and the method comprises administering to the subject an amount of the lipid binding protein-based complex that improves the subject's vision.

19. The lipid binding protein-based complex for use according to any one of claims 1 to 18, wherein the subject suffers from ocular lipid deposition, optionally wherein the ocular lipid deposition comprises corneal lipid deposition, retinal lipid deposition, eyelid lipid deposition or a combination thereof.

20. The lipid binding protein based complex for use according to any one of claims 1 to 19, wherein the lipid binding protein based complex is CER-001.

21. A lipid-binding protein-based complex for use according to any one of claims 1 to 20, wherein the lipid-binding protein-based complex is a carrier of one or more ophthalmic drugs, optionally wherein (I) the lipid-binding protein-based complex is CER-001, CSL-111, CSL-112, ETC-216, CER-522, delipidated HDL, Apomer or Cargomer and / or (II) the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

22. The lipid binding protein-based complex for use according to claim 21, wherein the one or more ophthalmic drugs comprise steroids, kinase inhibitors, angiotensin II receptor antagonists, aldose reductase inhibitors, immunosuppressants, carbonic anhydrase inhibitors, antimicrobial agents, antiviral agents, antihistamines, anti-inflammatory agents, prostaglandin analogs or combinations thereof.

23. The lipid binding protein-based complex for use according to claim 21 or 22, wherein the one or more ophthalmic drugs include dexamethasone palmitate, azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6-(3,5- dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, 2-methyl Sorbitol, sirolimus, cyclosporine, tacrolimus, acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, latanoprost, trovaprost, bimatoprost, or a combination thereof.

24. The lipid binding protein based complex for use according to any one of claims 21 to 23, wherein the one or more ophthalmic drugs comprises dexamethasone palmitate.

25. The lipid binding protein based complex for use according to any one of claims 21 to 23, wherein the one or more ophthalmic drugs comprises dexamethasone.

26. The lipid binding protein based complex for use according to any one of claims 21 to 23, wherein the one or more ophthalmic drugs comprises tacrolimus.

27. The lipid binding protein based complex for use according to any one of claims 1 to 26, wherein the method comprises peripheral administration of the lipid binding protein based complex, optionally by infusion.

28. The lipid binding protein-based complex for use according to claim 27, wherein the method comprises administering the lipid binding protein-based complex according to a dosing regimen comprising: (a) induction regimen; and / or (b) Consolidation programmes; and / or (c) maintenance program, Optionally, wherein the lipid binding protein based complex is CER-001.

29. The lipid binding protein based complex for use according to any one of claims 1 to 26, wherein the method comprises topically administering the lipid binding protein based complex.

30. The lipid binding protein based complex for use according to claim 29, wherein the method comprises intraocular administration of the lipid binding protein based complex.

31. The lipid-binding protein-based complex for use according to claim 30, wherein the method comprises administering the lipid-binding protein-based complex by intraocular injection, optionally wherein the intraocular injection is a suprachoroidal injection, a suprascleral injection, an intravitreal injection, a subconjunctival injection, a juxtabulbar injection, a peribulbar injection or a retrobulbar injection.

32. The lipid binding protein-based complex for use according to claim 29, wherein the method comprises administering the lipid binding protein-based complex via an implant, optionally wherein the implant is a biodegradable or non-biodegradable disc, sheet, plug, rod or pellet.

33. The lipid binding protein based complex for use according to claim 29, wherein the method comprises topically administering the lipid binding protein based complex.

34. The lipid binding protein based complex for use according to claim 33, wherein the lipid binding protein based complex is formulated as eye drops.

35. A composition which is an implant, optionally wherein the implant is a biodegradable or non-biodegradable disc, sheet, plug, rod or pellet, comprising a lipid binding protein-based complex and one or more ophthalmic drugs, optionally wherein (I) the lipid binding protein-based complex is CER-001, CSL-111, CSL-112, ETC-216, CER-522, delipidated HDL, Apomer or Cargomer and / or (II) one or more of the one or more ophthalmic drugs are (i) hydrophobic and / or (ii) poorly water soluble or water insoluble.

36. The composition of claim 35, wherein the lipid binding protein based complex is CER-001.

37. The composition of claim 35 or claim 36, wherein the one or more ophthalmic drugs comprise a steroid, a kinase inhibitor, an angiotensin II receptor antagonist, an aldose reductase inhibitor, an immunosuppressant, a carbonic anhydrase inhibitor, an antimicrobial agent, an antiviral agent, an antihistamine, an anti-inflammatory agent, or a combination thereof.

38. The composition of any one of claims 35 to 37, wherein the one or more ophthalmic drugs comprises dexamethasone palmitate, azithromycin, dexamethasone, difluprednate, estradiol, fluocinolone, fluorometholone, hydrocortisone, loteprednol etabonate, prednisolone, triamcinolone acetonide, rimexolone, spironolactone, axitinib, BMS-794833 (N-(4-((2-amino-3-chloropyridin-4-yl)oxy)-3-fluorophenyl)-5-(4-fluorophenyl)-4-oxo-1,4-dihydropyridine-3-carboxamide), cabozantinib, cediranib, dovitinib, lapatinib, lenvatinib, motesanib, nintedanib, olantinib, PD173074 (N-[2-[[4-(diethylamino)butyl]amino]-6 -(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl]-N'-(1,1-dimethylethyl)urea), pazopanib, regorafenib, sorafenib, tofacitinib, ZM323881 (5-((7-benzyloxyquinazolin-4-yl)amino)-4-fluoro-2-methylphenol), candesartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, 2-methylsorbitol, sirolimus, cyclosporine, tacrolimus, acetazolamide, brinzolamide, dorzolamide, ethoxazolamide, methazolamide, acyclovir, chloramphenicol, chlortetracycline, ciprofloxacin, fusidic acid, ganciclovir, norfloxacin, ofloxacin, tetracycline, zidovudine, levocabastine, bromfenac, diclofenac, indomethacin, nepafenac, or a combination thereof.

39. The composition of claim 38, wherein the one or more ophthalmic drugs comprises dexamethasone palmitate, optionally wherein the concentration of dexamethasone palmitate in the composition is 1 mg / ml.

40. The composition of claim 38, wherein the one or more ophthalmic drugs comprises dexamethasone.

41. The composition of claim 38, wherein the one or more ophthalmic drugs comprises an ophthalmic drug comprising tacrolimus.

42. The composition of any one of claims 35 to 41, which is a pharmaceutical composition further comprising one or more buffers, preservatives, excipients, diluents, or combinations thereof, optionally wherein the pharmaceutical composition is formulated as eye drops.

43. A composition which is an implant comprising a lipid binding protein-based complex, optionally wherein the lipid binding protein-based complex is CER-001, optionally wherein the implant is a biodegradable or non-biodegradable disc, sheet, plug, rod or pellet.

44. The composition of any one of claims 1 to 43 for use in a method of treating an eye disease in a subject.

45. A method of treating an eye disease in a subject, the method comprising: (a) administering to the subject an apolipoprotein AI ("ApoA-I") preparation comprising ApoA-I and one or more lipids, wherein the ApoA-I and the lipids are present in the form of a lipoprotein complex, wherein: (i) the subject suffers from cataract, uveitis, eye disease associated with lipid accumulation, lipid keratopathy, corneal dystrophy, corneal opacity, dry eye disease, blepharitis, inflammatory eye disease, macular edema (e.g., diabetic macular edema), macular degeneration, retinal detachment, eye tumor, fungal infection, viral infection, bacterial infection (e.g., bacterial conjunctivitis or trachoma), multifocal choroiditis, diabetic retinopathy, proliferative vitreoretinopathy (PVR), sympathetic ophthalmia, Vogt Koyanagi-Harada (VKH) syndrome, histoplasmosis, uveal spread, vascular occlusion, endophthalmitis, glaucoma, Stargardt's disease, ocular lipid deposition; and / or (ii) administering the formulation by suprachoroidal injection, suprascleral injection, implant, iontophoresis or electroporation.

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