Combination therapies for eye conditions or disorders

Abstract

Methods of ameliorating macular edema in a subject in need thereof; methods involve administering Edonentan or a form thereof and a VEGF inhibitor. Compositions having both Edonentan or a form thereof and a VEGF inhibitor.

Description

Attorney Docket No.: PFT-020WOCOMBINATION THERAPIES FOR EYE CONDITIONS OR DISORDERS CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Application No.63 / 756,778 (filed February 10, 2025) the contents of which are incorporated by reference herein in their entirety.SEQUENCE LISTING

[0002] The present specification makes reference to a Sequence Listing which has been submitted electronically in XML format and is incorporated by reference herein in its entirety. Said XML file, created on February 10, 2026, is named PFT-020WO_Sequence_Listing.xml and is 19,923 bytes in size.BACKGROUND

[0003] Diseases of the eye have an enormous impact on the quality of human life and yet remain largely elusive to effective treatment. It is estimated that an annual economic burden of over $100 billion results from vision loss, eye diseases, and vision disorders in the United States.

[0004] Macular edema is a condition where fluid accumulates in the macula, the central part of the retina responsible for sharp, central vision. This swelling arises from an imbalance between retinal fluid entry and exit mechanisms and is a nonspecific manifestation of underlying ocular diseases such as diabetic macular edema, age-related macular degeneration, or retinal vein occlusion. Patients affected by macular edema commonly experience symptoms such as metamorphopsia, micropsia, blurred vision, central scotoma, and reduced contrast or color sensitivity. Macular edema can cause blurred vision, distortion, and in severe cases, vision loss.

[0005] There remains a need to more effectively reduce the incidence of, treat, or otherwise ameliorate macular edema.SUMMARY

[0006] The present disclosure addresses this need with methods of ameliorating macular edema.1I PTS / 2OO316166.1Attorney Docket No.: PFT-020WG

[0007] In one aspect, provided are methods for ameliorating macular edema in a subject in need thereof, the method comprising a step of: co-administering to the subject (1) a sustained release bio-erodible device having a total amount of between about 150 pg and about 650 pg of a crystalline form of Edonentan, by implanting the bio-erodible device into the vitreous cavity of the subject’s eye and (2) a bispecific VEGF / angiopoietin 2 antibody, wherein the subject exhibits: (a) improved visual acuity as evidenced by an increase relative to a reference level in the Best Corrected Visual Acuity (BCVA) score by at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, or at least 18 Early Treatment Diabetic Retinopathy (ETDRS) letters, at four weeks after administration of the bispecific VEGF / angiopoietin 2 antibody, (b) a reduction relative to a reference level in at least 20 pm, at least 40 pm, at least 60 pm, at least 80 pm, at least 100 pm, at least 120 pm, at least 140 pm, or at least 150 pm in the Central Subfield Thickness (CST) as determined by optical coherence tomography, at four weeks after administration of the bispecific VEGF / angiopoietin 2 antibody, or (c) a visual field mean deviation (VF MD) slope of at least 0.50 dB / year, at least 0.75 dB / year, at least 1.00 dB / year, at least 1.25 dB / year, at least 1.50 dB / year, at least 1.75 dB / year, at least 2.00 dB / year, at least 2.25 dB / year, at least 2.50 dB / year as determined by a Frequency Doubling Perimetry (FDP) over a period of at least or about three months, at least or about four months, at least or about five months, or at least or about six months after implanting, wherein the reference level is a level determined by the same assay at a timepoint when the subject is determined to be experiencing macular edema.

[0008] In one aspect, provided are methods for ameliorating macular edema in a subject in need thereof, the method comprising co-administering to the subject (1) a first composition comprising Edonentan, a pharmaceutically acceptable salt thereof, a crystalline form thereof, or an amorphous form thereof, and (2) a second composition comprising a VEGF inhibitor.

[0009] In another aspect, provided are improvements to methods of treating a subject in need thereof with a VEGF inhibitor, the improvement comprising co-administering a first composition comprising Edonentan, a pharmaceutically acceptable salt thereof, a crystalline form thereof, or an amorphous form thereof.

[0010] In some embodiments, the subject is a mammal, e.g., a human.

[0011] In some embodiments, the subject is suffering from, at risk of developing, or diagnosed with an eye disease or disorder, such as an eye disease or disorder selected from the group consisting of age-related macular degeneration, diabetic retinopathy, eye tumor.2I PTS / 2OO316166.1Attorney Docket No.: PFT-020WGglaucoma, retinal vein occlusion, retinitis pigmentosa, retinoschisis, uveitis, and vitreomacular traction. In some embodiments, the eye disease or disorder is diabetic retinopathy.

[0012] In some embodiments, the subject has undergone an eye surgery or has experienced an injury to the eye.

[0013] In some embodiments, the subject exhibits a reduction in the central subfield thickness after co-administration, for example, a reduction which comprises a reduction of least 125 m at four weeks after co-administration

[0014] In some embodiments, the subject exhibits an improvement in visual acuity after co-administration. In some embodiments, the improvement in visual acuity comprises an increase of at least 8 Early Treatment Diabetic Retinopathy (ETDRS) letters on a Best Corrected Visual Acuity test at four weeks after co-administration.

[0015] In some embodiments, the subject exhibits reduced retinal neovascularization.

[0016] In some embodiments, co-administration comprises intravitreal administration of the first composition, for example, intravitreal administration by implanting a sustained release, bio-erodible device which comprises the first composition in the vitreous cavity of the eye.

[0017] In some embodiments, the bio-erodible device comprises a poly(lactic-co-glycolic acid) (PLGA) polymer. In some embodiments, the first composition comprises a total dose of about 200 pg, about 400 pg, or about 500 pg of Edonentan.

[0018] In some embodiments, administration comprises administering a composition, e.g., the first composition, at a regular interval, e.g., a regular interval of at least three months, about four months, or about six months.

[0019] In some embodiments, the VEGF inhibitor comprises a VEGF antibody, such as a monospecific antibody or a bispecific antibody, such as a VEGF / angiopoietin 2 antibody, e.g., faricimab. In some embodiments, the VEGF inhibitor comprises a fusion protein comprising a VEGF-R receptor. In some embodiments, the VEGF inhibitor comprises a small molecule.

[0020] In some embodiments, the first composition is administered separately from the second composition or the VEGF inhibitor. In some embodiments, the first composition is administered before the second composition or the VEGF inhibitor is administered.3I PTS / 2OO316166.1Attorney Docket No.: PFT-020WG

[0021] In some embodiments, the second composition or the VEGF inhibitor is administered before the first composition is administered. In some embodiments, wherein the first composition and second composition or VEGF inhibitor are administered at the same time.

[0022] In one aspect, provided are compositions comprising (1) Edonentan, a pharmaceutically acceptable salt thereof, a crystalline form thereof, or an amorphous form thereof, and (2) a VEGF inhibitor, wherein the Edonentan and VEGF inhibitors are present in amounts that are therapeutically effective in combination.BRIEF DESCRIPTION OF THE DRAWINGS

[0023] FIG. 1A depicts the Best Corrected Visual Acuity (BCVA) Early Treatment of Diabetic Retinopathy Study (ETDRS) letter score for a diabetic retinopathy patient at screening, at baseline, and at various timepoints (seven days, two months, three months, four months, five months, and six months) after the beginning of low-dose treatment with Edonentan delivered via Implant 1. The y-axis is marked at 50, 55, 60, 65, 70, 75, 80, and 85. At month three, 6 mg of faricimab-svoa (an anti-VEGF / anti-angiopoietin-2 bispecific antibody) was administered.

[0024] FIG. IB depicts the Central Subfield Thickness (CST) (pm) obtained by optical coherence tomography (OCT) for the same diabetic retinopathy patient at screening, at baseline, and at various timepoints (one month, two months, three months, four months, five months, and six months) after the beginning of low-dose treatment with Edonentan delivered via Implant 1. The y-axis is marked at 0, 50, 100, 150, 200, 250, 300, 350, 400, 450, and 500. At month three, 6 mg of faricimab-svoa (an anti-VEGF / anti-angiopoietin-2 bispecific antibody) was administered.

[0025] FIG. 2 depicts results from frequency Doubling Perimetry (FDP) measurements over time in the same diabetic retinopathy patient. The mean deviation is plotted on the y-axis against years on the x-acis. The y-axis is marked at -15, -10, -5, 0, and 5. The x-axis is marked at 0.0, 0.1, 0.2, 0.3, 0.4, and 0.5 The y-intercept of the line is calculated to be -9.0501, and the slope is calculated to be +2.9301.4I PTS / 2OO316166.1Attorney Docket No.: PFT-020WODETAILED DESCRIPTION OF CERTAIN EMBODIMENTSDefinitions

[0026] As used herein, the terms “about” or “approximately” when used herein in reference to a value, are used interchangeably and refer to a value that is similar to the referenced value. In general, those skilled in the art and familiar with the context will appreciate the relevant degree of variance encompassed by “about” in that context. For example, in some embodiments, the term "about" may encompass a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.

[0027] As used herein, the term “amorphous” refers to a solid material having no long range order in the position of its molecules. Amorphous solids are generally supercooled liquids in which the molecules are arranged in a random manner so that there is no well-defined arrangement, e.g., molecular packing, and no long range order. Amorphous solids are generally isotropic, i.e. exhibit similar properties in all directions and do not have definite melting points. For example, an amorphous material is a solid material having no sharp characteristic crystalline peak(s) in its X-ray power diffraction (XRPD) pattern (i.e., is not crystalline as determined by XRPD). Instead, one or several broad peaks (e.g., halos) appear in its XRPD pattern.

[0028] As used herein, the terms “baseline” refers to a level used for comparison. For example, a baseline level may refer to the level for the same indicator being measured before a treatment is administered, or at approximately the same time as the beginning of a treatment, in the same subject who is being evaluated at later timepoints.

[0029] As used herein, the terms “biodegradable,” and like terms generally describes a device and refers to a biologically assisted degradation process that the substance, e.g., polymer, making-up the device undergoes in a biological environment, such as within the body of a subject. Biodegradation encompasses within the scope the processes of absorption, dissolution, breaking down, degradation, assimilation, or otherwise removal of the device from the body, a biological environment. For example, a biodegradable polymer is liable to hydrolysis under physiological conditions due to the presence of hydrolytically and / or enzymatically susceptible functional groups.5I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO

[0030] As used herein, the term “bio-erodible” in reference to a material, e.g., a device or an implant, refers to a characteristic of the material in that the material erodes mechanically via a biological process, e.g., for polymer-based bio-erodible materials, the biological process may solubilize the polymer an enable absorption of the polymer into surrounding tissue.

[0031] As used herein, a “carrier” refers to a compound that facilitates the incorporation of a compound into cells or tissues. For example, without limitation, dimethyl sulfoxide (DMSO) is a commonly utilized carrier that facilitates the uptake of many organic compounds into cells or tissues of a subject.

[0032] As used herein, the terms “co-administer,” “administer in combination,” “combined administration,” and like terms means that two or more agents are administered to a subject at the same time or within an interval such that there may be an overlap of an effect of each agent on the patient. Thus, two or more agents that are administered in combination need not be administered together. In some embodiments, the two agents are administered within 180 days (e.g., within 180, 170, 160, 150, 140, 130, 120, 110, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, 5, 4, 3, 2, or 1 day(s)), within 28 days (e.g., with 14, 7, 6, 5, 4, 3, 2, or 1 day(s), within 24 hours (e.g., 12, 6, 5, 4, 3, 2, or 1 hour(s), or within about 60, 30, 15, 10, 5, or 1 minute of one another. In some embodiments, the administrations of the agents are spaced sufficiently closely together such that a combinatorial effect is achieved.

[0033] As used herein, a “diluent” refers to an ingredient in a composition that lacks pharmacological activity but may be pharmaceutically necessary or desirable which may be used (1) to increase the bulk of a potent drug whose mass is too small for manufacture and / or administration or (2) for the dissolution of a drug to be administered by injection, ingestion or inhalation. A common form of diluent in the art is a buffered aqueous solution such as, without limitation, phosphate buffered saline that mimics the composition of human blood.

[0034] As used herein, an “excipient” refers to an inert substance that is added to a p\ composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability, retarded dissolution etc., to the composition. A “diluent” is a type of excipient.

[0035] As used herein, the phrase “pharmaceutically acceptable” is used to describe a carrier, diluent, excipient, salt or composition that is safe and effective for its intended use and possesses the desired biological and pharmacological activity.6I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO

[0036] As used herein, the term “pharmaceutical composition” refers to a mixture of a compound or other agent with other chemical components, such as diluents or carriers, and is suitable for pharmaceutical use, e.g., administration to an organism.

[0037] As used herein, the term “polymer” encompasses both homopolymers (polymers having only one type of repeating unit) and copolymers (a polymer having more than one type of repeating unit). The term “biodegradable polymer” as used herein refers to a polymer or polymers, which degrade in vivo, under physiological conditions. The release of a therapeutic agent occurs concurrent with, or subsequent to, the degradation of a biodegradable polymer over time.

[0038] As used herein, the phrases “therapeutically effective amount” and “effective amount” are used interchangeably and refer to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result. A therapeutically effective amount may vary according to factors such as the type of disease e.g., disease state, age, sex, and / or weight of the individual, and the ability of a compound (or a composition thereof) to elicit a desired response in the individual. An effective amount may also be an amount for which any toxic or detrimental effects of a composition (e.g., a composition comprising Edonentan or a form thereof) are outweighed by therapeutically beneficial effects.

[0039] As used herein, the term “reference” describes a standard or control relative to which a comparison is performed, and a “reference level” refers to a level from such a standard or control. For example, in some embodiments, an agent, animal, subject, population, sample, sequence or value of interest is compared with a reference or control agent, animal, subject, population, sample, sequence or value. In some embodiments, a reference or control is tested and / or determined substantially simultaneously with the testing or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and / or comparison to a particular possible reference or control.

[0040] As used herein, the term “subject” refers to an organism, typically a mammal (e.g., a human). In some embodiments, a subject is suffering from or susceptible to a relevant disease, disorder or condition. In some embodiments, a subject displays one or more7I PTS / 2OO316166.1Attorney Docket No.: PFT-020WGsymptoms or characteristics of a disease, disorder or condition. In some embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition. In some embodiments, a subject is a patient. In some embodiments, a subject is a subject to whom diagnosis and / or therapy is and / or has been administered.Methods of ameliorating macular edema

[0041] Methods provided herein generally comprise a step of co-administering to a subject in need thereof (1) a composition comprising a therapeutically effective amount of Edonentan, a pharmaceutically acceptable salt thereof, a crystalline form thereof, or an amorphous form thereof, as described further herein, and (2) a composition comprising a VEGF inhibitor. As mentioned herein, “co-administering” means that the two compositions can be administered separately but close enough in time such that there may be an overlap of effect of each agent in the subject, or together. When the two agents are not administered together, the “timepoint” of co-administration is considered the time at which the latter of the two agents is administered.

[0042] Administration of one or more of the compositions may be achieved, in some embodiments, by use of a delivery device such as a sustained release bio-erodible device comprising a biodegradable polymer (e.g., Poly(lactic-co-glycolic acid) (PLGA)) as further described herein. As an example, when a sustained release formulation is used to administer at least one of the compositions, the other composition may be administered at a timepoint when an active agent is likely to still be released from the sustained release formulation.

[0043] In many embodiments, the subject is a mammal, e.g., a human.

[0044] In certain embodiments, the subject has or is at risk of having macular edema. For example, the subject may be suffering from, at risk of developing, or diagnosed with an eye disease or disorder associated with increased risk for macular edema. Non-limiting examples of such eye disorders condition include of age-related macular degeneration, diabetic retinopathy, eye tumor, glaucoma, retinal vein occlusion, retinitis pigmentosa, retinoschisis, uveitis, and vitreomacular traction. In some embodiments, the eye disease or disorder is diabetic retinopathy. In some embodiments, the subject has undergone a procedure or incident associated with increased risk of macular edema, e.g., an eye surgery or an injury (such as a traumatic injury) to the eye.8I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOAmeliorating a symptom

[0045] Provided methods result, in some embodiments, in amelioration of macular edema, and / or slowing progression of disease condition associated with macular edema.

[0046] “Ameliorating” a symptom may involve improving or relieving symptoms or preventing incidence or severity.

[0047] “Slowing disease progression” may include 1) slowing the rate of deterioration or worsening with respect to at least one aspect of a disease; 2) maintaining a function with respect to at least one aspect of a disease; and / or 3) improving a function or outcome with respect to at least one aspect of a disease. Thus, in some embodiments, slowing disease progression means extending the amount of time a subject with a given state of a disease reaches an advanced (e.g., more deteriorated, more severe, and or more complicated) state of the disease as compared to a reference level or as compared to the level from a comparable subject who has not been administered a composition in accordance with disclosed methods. The amount of time may be extended, in some embodiments, by 10%, by 15%, by 20%, by 25%, by 30%, by 35%, by 40%, by 45%, by 50%, by 55%, by 60%, by 65%, by 70%, by 75%, by 80%, by 85%, by 90%, by 95%, by 100%, by 125%, by 150%, by 175%, by 200%, by 250%, by 300%, by 350%, by 400%, by 450%, by 500%, or by more than 500%.

[0048] In some embodiments, provided methods result in the amelioration of (e.g., slowing or preventing) of vision loss (e.g., central and / or peripheral vision loss), which would normally be expected in a patient over time, the maintenance of vision (e.g., central and / or peripheral vision), or an improvement of vision (e.g., central and / or peripheral vision), over a period of time. For example, in some embodiments, methods result in maintained or improved peripheral vision, maintained or improved contrast sensitivity, maintained or improved low contrast visual acuity, and / or maintained or improved visual acuity over a period of least one, at least two, at least three, at least four, at least five, or at least six months after co-administration.

[0049] The ability to see delicate objects does not guarantee the ability to see large or medium-sized objects of low contrast. Contrast sensitivity (CS) is a measure of visual ability to distinguish an object from its background. Contrast sensitivity function (CSF) acquired with the quick CSF (qCSF) is a metric that incorporates both visual acuity (e.g., spatial resolution) and CS.9I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO

[0050] Contrast sensitivity can be evaluated using the qCSF (quick contrast sensitivity function) method on the AST Platform (Adaptive Sensory Technology, San Diego, CA, USA), a computerized method for evaluating the contrast thresholds over a wide range of contrast (0.002% - 100%) and spatial frequency (approximately 1 to 27 cycles per degree (CPD)). Subjects can be asked to read presented optotypes, and their responses can be registered (as correct, incorrect, or optotype not seen) on a tablet computer by a trained study nurse. All participants were tested monocularly for approximately 2 minutes. Sensitivities at various spatial frequencies (e.g., 1.5, 3.0, 5.0, or 6.0 CPD) can serve as a metric of constrast sensitivity function and used for statistical analyses. Low luminance low contrast visual acuity can be assessed in reduced light conditions, using, e.g., a neutral density filter.

[0051] In some embodiments, provided methods result in a reduction in the central subfield thickness after co-administration, for example, a reduction of at least 125 pm, at least 130 pm, at least 135 pm, at least 140 pm, at least 145 pm, or at least 150 pm at four weeks after co-administration.

[0052] For example, in some embodiments, provided methods result in maintenance or improvement of the visual field, e.g., an improvement represented by an increase of at least 0.25 dB, at least 0.50 dB, at least 0.75 dB, at least 1.00 dB, at least 1.25 dB, at least 1.50 dB, at least 1.75 dB, or at least 2.00 dB in the visual field mean deviation (VF MD) as determined by a Humphrey Field Analyser at or for three months after co-administration. In some embodiments, provided methods result in an improvement represented by an increase of at least 0.25 dB, at least 0.50 dB, at least 0.75 dB, at least 1.00 dB, at least 1.25 dB, at least 1.50 dB, at least 1.75 dB, or at least 2.00 dB in the visual field mean deviation (VF MD) as determined by a Humphrey Field Analyser at or for six months after co-administration.

[0053] For example, in some embodiments, provided methods result in maintenance or improvement of contrast sensitivity, e.g., no change relative to baseline, or an improvement represented by an increase of at least 0.25 dB, at least 0.50 dB, at least 0.75 dB, at least 1.00 dB, at least 1.25 dB, at least 1.50 dB, at least 1.75 dB, or at least 2.00 dB in the quick contrast sensitivity function (qCSF) method on the Adaptive Sensory Technology (AST) Platform at 1.5 CPD, 3.0 CPD, or 6.0 CPD at or for three months, six months, eight weeks, or twenty weeks, after co-administration.

[0054] In some embodiments, provided methods result in amelioration of impaired blood flow to the optic nerve head, e.g., by increasing optic nerve head capillary blood flow. In10I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOsome embodiments, provided methods result in an increase in the Mean Blur Rate (MBR) as determined by laser speckle flowgraphy. For example, the increase in the MBR can be, in some embodiments, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, or at least 30% over a baseline level. By “baseline level,” it is meant the level in the same subject at the time of administration or before administration, or a similar comparable reference level, such as a reference level from a control subject.

[0055] In some embodiments, provided methods result in an increase in retinal nerve fiber layer (RNFL) thickness after co-administration of the composition, e.g., as determined by a technique such as optical coherence tomography. For example, the increase in the RNFL thickness can be, in some embodiments, at least 0.5 pm at least 1.0 pm, at least 1.5 pm, at least 2.0 pm, at least 2.5 pm or at least 3.0 pm from a baseline level at or for three months after co-administration.

[0056] In some embodiments, provided methods result in the slowing or preventing of a vision-threatening complication, such as macular edema.

[0057] In some embodiments, provided methods result in maintenance or improvement of visual acuity, e.g., maintenance of visual acuity over a period of at least one, at least two, at least three, at least four, at least five, or at least six months after co-administration. For example, when visual acuity is assessed by Best Corrected Visual Acuity (BCVA), in some embodiments, provided methods result in a score improvement of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen, at least nineteen, or at least twenty Early Treatment Diabetic Retinopathy (ETDRS) letters on the BCVA at a given timepoint after co-administration, for example, a timepoint of at least one, at least two, at least three, at least four, at least five, or at least six months after co-administration.

[0058] In some embodiments, methods result in reduced retinal neovascularization. For example, reduced neovascularization can be determined by imaging methods.

[0059] In some embodiments, slowing disease progression comprises maintaining or improving a clinical status or score associated with the disease, for example, a clinical status or score associated with severity of macular edema or a disease associated with macular edema.11I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO

[0060] In some embodiments, the clinical score is maintained or improved over a period of time after co-administration, e.g., at least one, at least two, at least three, at least four, at least five, or at least six months after co-administration. In some embodiments, the clinical score is maintained or improved over a period of at least three, at least four, or at least six months after the co-administration.Compositions comprising Edonentan and / or forms thereofEdonentan

[0061] Edonentan is a highly selective and very potent endothelin A receptor antagonist with the chemical name: N-[[2'-[[(4,5-dimethyl-3-isoxazolyl)amino]sulfonyl]-4-(2-oxazolyl)[l,l'-biphenyl]-2-yl]methyl]-N,3,3-trimethylbutanamide. Edonentan has a molecular weight of 536.6 g / mol and the following structure:Edonentan (Compound I)

[0062] Methods of preparing Edonentan are well known to a person of skill in the art.Suitable methods are disclosed, for example, in U.S. Patent No. 6,043,265.

[0063] Compositions used in methods disclosed herein can include therapeutically effective amount of Edonentan or of any form thereof, such as a pharmaceutically acceptable salt for, a crystalline form, an amorphous form, or a combination of any of the foregoing.

[0064] Crystalline forms of Edonentan are substantially more stable compared to the amorphous form of Edonentan. For example, crystalline “Form 4” of Edonentan, disclosed in International Patent Publication WO 2021 / 158663 Al, may be stored under practical and economical storage conditions, while retaining physical properties so that it may be manufactured into a dosage form. A given crystalline form (e.g., Form 1, Form 2, or Form 3 disclosed in WO 2021 / 158663 Al) may be converted to another crystalline form (e.g., Form 4 disclosed in WO 2021 / 158663 Al), as further explained in WO 2021 / 158663 Al.12I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO

[0065] In some embodiments, a given crystalline form, together with any other forms of Edonentan present in the composition, are present in amounts that are therapeutically effective together. In some embodiments, the composition comprises a therapeutically effective amount of a given crystalline form. In some embodiments, the composition comprises crystalline Form 4 of Edonentan (as described in WO 2021 / 158663 Al).

[0066] In some embodiments, the composition comprises a hydrate form of crystalline Edonentan, e.g., Edonentan • (H2O)m, where m is a fractional or whole number between about 0 and about 4 inclusive. For example, anhydrate or monohydrate forms of crystalline Edonentan may be included in the composition. In some embodiments, a crystalline form of Edonentan may have a water level of about 1 to 10% by weight (e.g., 3 to 9% or 5 to 8% by weight).Crystalline forms

[0067] In certain embodiments, compositions comprise a crystal form of the compound of Formula I:

[0068] In some embodiments, the compound of Formula I is in an anhydrous crystalline form (Form 4), having an X-ray powder diffraction pattern as determined by Cu Ka radiation comprising at least three characterization peaks, in terms of 29, selected from peaks at 5.6+0.2°, 11.4+0.2°, 14.4+0.2°, 15.7+0.2°, 16.8+0.2°, 17.7+0.2°, 19.3+0.2°, 21.1+0.2°, 21.9+0.2°, 23.9+0.2°, and 24.6+0.2°.

[0069] In some embodiments, Form 4 has an X-ray powder diffraction pattern as determined by Cu Ka radiation comprising the following peaks expressed in terms of diffraction angles (29): 5.6+0.2°, 11.4+0.2°, 17.7+0.2°, 19.3+0.2°, 21.1+0.2°, and 21.9+0.2°. In some embodiments, Form 4 has an X-ray powder diffraction pattern as determined by Cu Ka radiation comprising the following peaks expressed in terms of diffraction angles (29):5.6+0.2°, 11.4+0.2°, 17.7+0.2°, 19.3+0.2°, and 21.9+0.2°. In some embodiments, Form 4 has 13I PTS / 2OO316166.1Attorney Docket No.: PFT-929WOan X-ray powder diffraction pattern as determined by Cu Ka radiation comprising the following peaks expressed in terms of diffraction angles (29): 11.4+9.2°, 17.7+0.2°, and 19.3+0.2°. In some embodiments, Form 4 has an X-ray powder diffraction pattern as determined by Cu Ka radiation comprising the following peaks expressed in terms of diffraction angles (29): 5.6+9.20, 11.4+0.2°, 14.4+0.2°, 15.7+0.2°, 16.8+0.2°, 17.7+0.2°, 19.3+0.2°, 21.1+0.2°, 21.9+0.2°, 23.9+0.2°, and 24.6+0.2°.

[0070] In some embodiments, Form 4 has a Tmof about 163 °C by DSC analysis.

[0071] In some embodiments, at least 90% by weight (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by weight) of the total amount of the compound of Formula I in the composition is in the form of Form 4.

[0072] In certain embodiments, the compound of Formula I is in an anhydrous crystalline form (Form 1), wherein the anhydrous crystalline Form 1 has an X-ray powder diffraction pattern as determined by Cu Ka radiation comprising at least three characterization peaks, in terms of 29, selected from peaks at 6.3+0.2°, 7.5+0.2°, 11.7+0.2°, 15.1+0.2°, and 17.3+0.2°; and 90% by weight or more of the total amount of the compound of Formula I in the composition is in the form of Form 1.

[0073] In certain embodiments, the compound of Formula I is in a monohydrate crystalline form (Form 2), wherein the monohydrate crystalline Form 2 has an X-ray powder diffraction pattern as determined by Cu Ka radiation comprising at least three characterization peaks, in terms of 29, selected from peaks at 9.6+0.2°, 10.4+0.2°, 19.6+0.2°, 19.7+0.2°, 22.0+0.2°, 22.9+0.2°, and 23.7+0.2°; and 90% by weight or more of the total amount of the compound of Formula I in the composition is in the form of Form 2.

[0074] In certain embodiments, the compound of Formula I is in an anhydrous crystalline (Form 3), wherein the anhydrous crystalline Form 3 has an X-ray powder diffraction pattern as determined by Cu Ka radiation comprising at least three characterization peaks, in terms of 29, selected from peaks at 7.8+0.2°, 9.0+0.2°, 11.6+0.2°, 15.8+0.2°, and 19.1+0.2°; and 90% by weight or more of the total amount of the compound of Formula I in the composition is in the form of Form 3.

[0075] As used herein, the term “amorphous” refers to a solid material having no long range order in the position of its molecules. Amorphous solids are generally supercooled liquids in which the molecules are arranged in a random manner so that there is no well-defined arrangement, e.g., molecular packing, and no long range order. Amorphous solids are14I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOgenerally isotropic, i.e. exhibit similar properties in all directions and do not have definite melting points. For example, an amorphous material is a solid material having no sharp characteristic crystalline peak(s) in its X-ray power diffraction (XRPD) pattern (i.e., is not crystalline as determined by XRPD). Instead, one or several broad peaks (e.g., halos) appear in its XRPD pattern.

[0076] Hydrate forms of crystalline Edonentan are contemplated, e.g., Edonentan • (H2O)m, where m is a fractional or whole number between about 0 and about 4 inclusive. For example, contemplated herein are anhydrate or monohydrate forms of crystalline Edonentan. In an embodiment, a disclosed crystalline form of Edonentan may have a water level of about 1 to 10% by weight (e.g., 3 to 9% or 5 to 8% by weight).Other components

[0077] In some embodiments, compositions comprise a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.

[0078] Compositions for use in accordance with the presently disclosed methods can be administered to a human patient per se, or in compositions where they are mixed with other active ingredients (e.g., as in a combination therapy), or carriers, diluents, excipients or combinations thereof. Proper formulation may depend upon the route of administration chosen. Techniques for formulation and administration of the compounds described herein are known to those skilled in the art.

[0079] Compositions may be manufactured e.g., by means of mixing, dissolving, granulating, levigating, emulsifying, encapsulating or entrapping processes. Compositions for use in methods disclosed herein may also be formulated for particular delivery mechanisms and / or routes of administration, e.g., as further described herein.

[0080] In some embodiments, the composition is an ophthalmic preparation comprising a therapeutically effective amount of Edonentan or a form thereof. As used herein, an “ophthalmic preparation” refers to a specialized dosage form designed to be instilled onto the external surface of the eye (topical), administered inside (intraocular) or adjacent (periocular) to the eye or used in conjunction with an ophthalmic device. In some embodiments, the ophthalmic preparation is in the form of a solution, suspension, or an ointment. In other embodiments, the ophthalmic preparation is in the form of a gel, a gel-forming solution, an15I PTS / 2OO316166.1Attorney Docket No.: PFT-020WGocular insert, a micro / nanoparticle preparations for topical or preferably intravitreal injection, or an implant, e.g., as further described herein.

[0081] In some embodiments, the ophthalmic preparation comprises a preservative.Examples of suitable preservatives include, but are not limited to, cationic wetting agents (e.g., benzalkonium chloride), organic mercurials (e.g., phenylmercuric nitrate, phenylmercuric acetate), organic acids or their esters (e.g., sorbic acid, esters of p-hydroxybenzoic acid such as methyl hydroxybenzoate, propylhydroxybenzoate), and alcohol substitutes (e.g., chlorobutanol, phenylethanol). The preservative can be present in the ophthalmic preparation in an amount in the range of about 0.002 % w / v to about 0.5 % w / v (e.g., 0.01 - 0.25 % w / v). The ophthalmic preparation can further comprise a preservative aid. Examples of suitable preservative aid include, but are not limited to, ethylenediaminetetraacetic acid (EDTA).

[0082] In some embodiments, the ophthalmic preparation comprises one or more additional excipients or agents to impart viscosity or lubrication, stabilize the active ingredients against decomposition, increase solubility of an active or inactive ingredient, adjust tonicity, or act as solvent. Examples of excipients or agents for imparting viscosity or lubrication include, but are not limited to, hypromellose, carbomer 974P, hydroxyethyl cellulose (HEC), polyvinyl alcohol, sodium hyaluronate, sodium carboxymethyl cellulose, Carbopol 940, hydroxypropylmethyl cellulose (HPMC), poloxamer, xyloglucan, alginic acid, sodium alginate, gellan gum, cellulose acetate phthalate, and xantham gum. Examples of excipients or agents as stabilizers include, but are not limited to, sodium bisulfite, sodium metabisulfite, sodium thiosulfate, and sodium sulfate / sulfuric acid, which can act as antioxidants. Examples of excipients or agents as solubilizers include, but are not limited to, providone, glycerol, polyethylene glycol (PEG), polypropylene glycol (PPG), PEG- stearate, poloxamer 407, tyloxapol, polysorbate 80, creatinine, cyclodextrin, and castor oil. Examples of excipients or agents for adjusting tonicity include sodium chloride, potassium chloride, calcium chloride dehydrate, magnesium chloride hexahydrate, sugars (e.g., sucrose, maltose, dextrose, etc.), glycerin, propylene glycol, mannitol, ascorbic acid, and acetylcysteine.

[0083] In some embodiments, the ophthalmic preparation comprises one or more buffers to adjust pH. Examples of buffers for adjusting pH include, but are not limited to, sodium citrate, monobasic sodium phosphate, dibasic sodium phosphate, boric acid, hepatahydrate, sodium acetate trihydrate, sodium citrate dihydrate, histidine, and phosphate buffered saline16I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO(PBS). The resulting composition can have a pH value of 5.0-8.5 (e.g., 5.0-6.0, 5.2-5.8, 6.0-8.0, 6.6-7.8, 6.2-8.2, and 6.2-7.5)

[0084] In some embodiments, the ophthalmic preparation comprises one or more surfactants. Non-limiting examples of surfactants include sorbitan ether esters of oleic acid (e.g., polysorbate or Tween 20 and 80) and tyloxapol.Vascular endothelial growth factor (VEGF) inhibitors and compositions thereof

[0085] In some embodiments, the VEGF inhibitor (an agent that blocks the activity of VEGF) binds to VEGF-A. Any of a variety of VEGF inhibitors may be used, including small molecule inhibitors, antibody-based inhibitors, and / or fusion protein “traps” which contain VEGF-binding elements. For example, antibody-based inhibitors of VEGF include both monospecific antibodies (e.g., bevacizumab, ranibizumab, and brolucizumab (a single chain antibody fragment)) and bispecific antibodies which are capable of binding VEGF (e.g., VEGF-A) and another antigen (e.g., faricimab, which is a VEGF / angiopoietin 2 bispecific antibody). Receptor decoy-based inhibitors include fusion proteins such as, without limitation, aflibercept and conbercept.

[0086] Compositions generally comprises a vascular endothelial growth factor (VEGF) inhibitor (e.g., as described herein) and a pharmaceutically acceptable excipient, as further described herein.

[0087] Characteristic sequences of certain VEGF antibodies and VEGF / angiopoietin 2 bispecific antibodies are known in the art. For example, below are characteristic sequences of an VEGF / angiopoietin 2 bispecific antibody. In some embodiments, the VEGF / angiopoietin 2 bispecific antibodu comprises (1) an anti- VEGF binding region which comprises an immunoglobulin heavy chain variable domain (anti- VEGF Vn) comprising complementaritydetermining regions CDR-H1, CDR-H2, and CDR-H3 having the sequences of SEQ ID NOs: 1, 2, and 3, respectively, and an immunoglobulin light chain variable domain comprising complementarity-determining regions CDR-L1, CDR-L2, and CDR-L3 (anti- VEGF VL) having the sequences of SEQ ID NOs: 4, 5, and 6, respectively, and (2) an anti-angiopoietin 2 binding region which comprises an immunoglobulin heavy chain variable domain comprising complementarity-determining regions CDR-H1, CDR-H2, and CDR-H3 (anti-angiopoietin 2 VH) having the sequences of SEQ ID NOs: 9, 10, and 11, respectively, and an immunoglobulin light chain variable domain (anti-angiopoietin 2 VL) comprising17I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOcomplementarity-determining regions CDR-L1, CDR-L2, and CDR-L3 having the sequences of SEQ ID NOs: 12, 13, and 14, respectively. In some embodiments, the anti-VEGF VH and anti-VEGF Vrare also characterized by amino acid sequences having at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to that of SEQ ID NOs: 9 and 10, respectively. In some embodiments, the anti-angiopoietin 2 VH and anti-angiopoietin 2 VL are also characterized by amino acid sequences having at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to that of SEQ ID NOs: 15 and 16, respectively. In some embodiments, the VEGF / angiopoietin 2 bispecific antibody comprises the aforementioned CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences and is also composed of four polypeptide chains: (1) a first polypeptide chain characterized by an amino acid sequence that is at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to that of SEQ ID NO: 17; (2) a second polypeptide chain characterized by an amino acid sequence that is at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to that of SEQ ID NO: 18; (3) a third polypeptide chain characterized by an amino acid sequence that is at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to that of SEQ ID NO: 19; and (4) a fourth polypeptide chain characterized by an amino acid sequence that is at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to that of SEQ ID NO: 19.Exemplary VEGF / angiopoietin 2 bispecific antibody sequences18I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO19TS / 200316166.1Attorney Docket No.: PFT-020WO

[0088] Appropriate dosages of VEGF inhibitors can be determined by the skilled person in the art.Delivery mechanisms, administration, and dosing

[0089] In some embodiments, administration of one or both compositions comprises local administration, e.g., via a topical ophthalmic formulation and / or via injection of the20I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOcompounds or compositions directly to the ocular tissue, for example, in a depot or sustained release formulation. Administration can be, e.g., intravitreal, suprachoroidal, periocular, topical, via subconjunctival injection of a formulation, via use of an implant technology, or any combination of the foregoing. In some embodiments, administration of one or both compositions comprises intravitreal administration.

[0090] In some embodiments, provided are compositions or delivery devices that contain both agents (Edonentan or a form thereof and a VEGF inhibitor), with the two agents being present in amounts that are therapeutically effective when the two agents act together. The amounts of each agent may be, or may not be, therapeutically effective as a monotherapy.

[0091] In some embodiments, administration of the Edonentan (or form thereof) composition is achieved using a sustained release formulation or device which releases the Edonentan or form thereof over a period of time. In some embodiments, the sustained release formulation or device releases Edonentan or form thereof over a period of at least one month, at least two months, at least three months, at least four months, at least five months, or at least six months. In some embodiments, the sustained release formulation or device releases Edonentan or form thereof over a period of about one month, about two months, about three months, about four months, about five months, or about six months.

[0092] For example, liposomal preparations can be used to slowly release the Edonentan or form thereof, sustaining the desired pharmacological effects. Alternatively or additionally, polyvinyl alcohol nanoparticles can be prepared by well-known methods to afford a sustained or extended release-formulation for topical or intra-ocular applications.

[0093] In some embodiments, administration, e.g., intravitreal administration, is achieved via a sustained release bio-erodible device, such as an intravitreal bio-erodible device as described in International Patent Application No. WO 2022 / 232588 Al. The bio-erodible device (which may be referred to herein as a “bio-erodible implant,” “implant,” or like terms) generally comprises Edonentan or a form thereof within a biodegradable substance, e.g., a biodegradable polymer.

[0094] Suitable polymeric materials or compositions for use in bio-erodible device include those materials which are biocompatible with the eye so as to cause no substantial interference with the functioning or physiology of the eye. Such polymeric materials may be also be biodegradable, e.g., a biodegradable polymer.21I PTS / 2OO316166.1Attorney Docket No.: PFT-020WGBiodegradable polymers

[0095] In some embodiments, the biodegradable polymer is a PLGA (poly(lactic-co-glycolic acid)), a copolymer of polylactic acid (PLA) and polyglycolic acid (PLG). PLGA polymers are known to degrade via backbone hydrolysis (bulk erosion), and the final degradation products are lactic and glycolic acids, which are non-toxic and considered natural metabolic compounds. Lactic and glycolic acids are eliminated safely via the Krebs cycle by conversion to carbon dioxide and water. Biocompatibility of PLGA polymers have been examined in both non-ocular and ocular tissues of animals and humans, and findings indicate that these polymers are well tolerated.

[0096] PLGA is synthesized through random ring-opening co-polymerization of the cyclic dimers of glycolic acid and lactic acid. Successive monomeric units of glycolic or lactic acid are linked together by ester linkages. The ratio of lactide to glycolide can be varied, altering the biodegradation characteristics of the product. By altering the ratio, it is possible to tailor the polymer degradation time. Importantly, drug release characteristics are affected by the rate of biodegradation, molecular weight, and degree of crystallinity in drug delivery systems. By altering and customizing the biodegradable polymer matrix, the drug delivery profile can be changed.

[0097] Examples of suitable PLGA polymers include, but are not limited to, the RESOMER® Product line from Evonik Industries identified as RG502, RG502H, RG503, RG503H, RG504, RG504H, RG505, RG653H, RG750S, RG752H, RG752S, RG753H, RG753S, RG755S, RG756S, RG757S, and RG858S. In some embodiments, the biodegradable polymer comprises a poly(lactic-co-glycolic acid) (PLGA), wherein the PLGA is selected from the group consisting of RG502, RG503, RG752S, RG753S, RG755S, RG756S, and RG858S. Compositions of each of the aforementioned PLGA polymers are known to those skilled in the art.

[0098] In some embodiments, the bio-erodible comprises at least two types of PLGA. In some embodiments, the biodegradable polymer comprises at least three types of PLGA (e.g., three to six types of PLGA, three types of PLGA, four types of PLGA, or five types PLGA).

[0099] In some embodiments, the bio-erodible device comprises at least two PLGA, wherein each PLGA is independently selected from the group consisting of RG502, RG502H, RG503, RG503H, RG504, RG504H, RG505, RG653H, RG750S, RG752H, RG752S, RG753H, RG753S, RG755S, RG756S, RG757S, and RG858S.22I PTS / 2OO316166.1Attorney Docket No.: PFT-020WG

[0100] PLGA copolymers can be an ester- terminated PLGA copolymer, as identified by the terminal ‘S’ in the product name, or an acid-terminated PLGA copolymer, as identified by the terminal ‘H’ in the product name. Inherent viscosities of PLGA copolymers range from approximately 0.14 to approximately 1.7 dL / g when measured at 0.1% w / v in CHCh at 25 °C. with an Ubbelohde size 0c glass capillary viscometer. In some embodiments, polymers have a molar ratio of polyD,L-lactide (PLA) to polyglycolide (PLG) from approximately 50:50 to approximately 85:15, such as, but not limited to, 50:50, 65:35, 75:25, and 85:15. In some embodiments, the PLGA has a molar ratio of PLA and PLG of about 65:35.

[0101] Other examples of PLGA polymers include those produced by Lakeshore Biomaterials identified as, but not limited to, DLG 1A, DLG 3 A, or DLG 4A. Such DLG polymers include both acid (A) and ester (E) terminated polymers with inherent viscosities ranging from approximately 0.0.5 to approximately 1.0 dL / g when measured at 0.1% w / v in CHCh at 25° C. with an Ubbelohde size 0c glass capillary viscometer. Example polymers used in various embodiments of the disclosure may include variation in the mole ratio of D,L-lactide to glycolide from approximately 1:99 to approximately 99:1, such as, but not limited to, 50:50, 65:35, 75:25, and 85:15.

[0102] RESOMERS® identified by an “RG” or “DLG” in the product name, such as RG752S, are poly(D,L-lactide-co-glycolide) (PLGA) having the general structure (Compound II):(Compound II)

[0103] DLG can be synthesized with various D,L-lactide-glycolide ratios. In some embodiments, DLG, such as 1A, with an inherent viscosity of approximately 0.05 to approximately 0.15 dL / g is used. In some embodiment, DLGs, such as 2A, with an inherent viscosity of approximately 0.15 to approximately 0.25 dL / g is used.

[0104] Additional examples of PLGA polymers include the Viatel™ line of products supplied by Ashland (Wilmington, Delaware), such as the Viatel™ DLG5002A (Poly(D,L- 23I PTS / 2OO316166.1Attorney Docket No.: PFT-020WGLactide-co-Glycolide) acid endcap LG 50:50), Viatel™ DLG5003A (Poly(D,L-Lactide-co-Glycolide) acid endcap LG 50:50), Viatel™ DLG5005A (Poly(D,L-Lactide-co-Glycolide) acid endcap LG 50:50), Viatel™ DLG5005E (Poly(D,L-Lactide-co-Glycolide) ester endcap LG 50:50), Viatel™ DLG7505A (Poly(D,L- Lactide-co-Glycolide) acid endcap LG 75:25), Viatel™ DLG8503E (Poly(D,L-Lactide-co-Glycolide) ester endcap LG 85:15), and Viatel™ (Poly(D,L-Lactide-co-Glycolide) ester endcap LG 85:15). (The ratios after “LG” signify the ratios of D,L-lactide to glycolide.)

[0105] Further examples of PLGA polymers include the Expansorb® line of products from SEQUENS (Ecully, France), such as Expansorb® PLGA 45:55, Expansorb® PLGA 50:50, Expansorb® PLGA 50:50 PEG, Expansorb® PLGA 55:45, Expansorb® PLGA 55:45-Glu, Expansorb® PLGA 50:50, Expansorb® PLGA 75:25, Expansorb® PLGA 75:25 PEG, Expansorb® PLGA 85:15, Expansorb® PLGA 90:10, and Expansorb® PLGA 95:5.Volumes

[0106] The volume that can be injected to a human eye at one time is about 50 to about 90 pL through the intravitreal route, up to about 450 pL through a subretinal route, and up to about 200 pL via suprachoroidal routes. The needle used in these routes is typically 27 to 30 G in size. The dose depends on the concentration that can be formulated to fit this volume, potency, target efficacy and pharmacokinetic profile for each indication. Generally, the injections to the eye will not be administered at a frequency greater than once per month per eye. In some embodiments, when a composition as described herein is administered in topical form (e.g., eye drop), the volume administered to a human eye at one time can be about 50 pL (the concentration of the eye drop can be about 5 mg / mL).Dosing

[0107] In some embodiments, a formulation, e.g., an intravitreal formulation will comprise a dose of a composition comprising an Edonentan crystalline form in the range of about 1 pg to about 1 mg. A first exemplary formulation comprises about 1 pg to about 1 mg of a composition comprising an Edonentan crystalline form, about 10 mM histidine HC1, about 10% a,a-trehalose dihydrate, and about 0.01% polysorbate 20. A second exemplary formulation comprises about 1 pg to about 1 mg of a composition comprising an Edonentan24I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOcrystalline form, about 10 mM sodium phosphate, about 40 mM sodium chloride, about 0.03 % polysorbate 20, and about 5% sucrose.

[0108] In some embodiments, a total dose of about 100 ptg, about 150 ptg, about 200 ptg,about 250 ptg, ab out 300 ptg, about 350 ptg, about 400 ptg, about 450 ptg, about 500 ptg, about 550 ptg, about 600 ptg, about 650 ptg, about 700 ptg, about 750 ptg, or about 800 ptgEdonentan is administered to the subject, e.g., via a sustained release formulation, e.g., a bio-erodible device, such as a bio-erodible device for intravitreal administration. In certain embodiments, the total dose com prises a therapeutically effective dose of the Edonentan or form thereof. In some embodiments in which a bio-erodible device is used, the total dose is contained in a single bio-erodible device. In some embodiments in which a bio-erodible device is used, the total dose is contained in multiple bio-erodible device, e.g., two bio-erodible devices, which are delivered together to t he subject at the appropriate location in or around the eye, e.g., into the vitreous cavity.

[0109] In some embodiments, the composition is administered at a regular interval, e.g., at regular intervals of at least one, at least two, at least three, at least four, at least five, at leastsix months. In some embodiments, the regular interval is about one, about two, about three, about four, about five, or about six months. For example, in embodiments in which a bio-erodible device (e.g., an intravitreal bio-erodible device) is implanted into a subject, the subject may be re-implanted with another bio-erodible device at regular intervals.Exemplary formulations of compositions Edonentan (or a form thereof)

[0110] By way of non-limiting example, Tables 1 and 2 provide examples of sustained delivery formulations and implants.Table 1. Examples of Edonentan Containing Sustained Delivery Formulations (1 -7) for the production of film disks25I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOTable 2. Examples of Edonentan Containing Sustained Delivery Formulations for the production of implants26I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOEXAMPLES EXAMPLE 1: Compound Physicochemical and Biochemical Characterization

[0111] Provided in Table 3 below are physicochemical and biochemical data for Edonentan.Table 3. Edonentan physicochemical and biochemical characterization<<aThe data are from the amorphous form.bCalculated property that considers surface charge distributions (mainly O and N).Compounds with a PSA around 90 or below would be predicted to cross the blood-brain barrier.27I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO

[0112] In the above table, the physicochemical data, e.g., solubility, were obtained following standard protocols known in the field (see, e.g., Reis et al., Mini Rev Med Chem., 2010, 10(11): 1071-6; Avdeef et al., Expert Opin Drug Metab Toxicol., 2005, l(2):325-42; Bharate et al., Comb Chem High Throughput Screen., 2016, 19(6):461-9; and Jain et al., J Pharm Biomed Anal., 2013, 86:11-35.); and the biochemical data, i.e., potency for ETA / ETB, were obtained following the protocols known in the field (see, e.g., Kirkby et al., Br J Pharmacol., 2008, 153(6): 1105-19; and Maguire et al., Br J Pharmacol., 2014, 171(24):5555-72.).EXAMPLE 2: Formulation of Edonentan for Intravitreal Use in Rabbit

[0113] An appropriate amount of Edonentan is dissolved in neat PEG400, followed by addition of a 15% CD (HP-|3-cyclodextrin) solution. The final concentration of PEG400 is measured to be 20%. Target concentrations are 5 mg / ml and 0.5 mg / ml based on the amount of Edonentan. The resulting solution is filtered using a 0.25 micron filter.EXAMPLE 3: Effects of Edonentan and Enodthelin 1 (ET-1) in a Rabbit Model

[0114] Adult, male Dutch-belted rabbits were given a 20 pl intravitreal injection (IVT) of 0.5 pg of Endothelin 1 ( ET-1) followed by a 20 pl intravitreal injection of 10-100 pg Edonentan given 30 min after the ET-1 administration. IOP, optical coherence tomography -angiography (OCT-A), and fluorescein angiograms (FA) were performed at pre-specified time points (30, 45, 60, and 75 min) following ET-1 and Edonentan administration to assess retinal blood flow changes induced by ET-1 + / - Edonentan.

[0115] Based in optical coherence tomography - angiography(OCT-A), ET-1 administration effectively induced a clear vasoconstriction in the retinal vascular beds within 45 min. Based on fluorescein angiography, the effect of ET-1 was then reversed with 10 pg of Edonentan administration within 90 min (60 min after Edonentan administration).28I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOEXAMPLE 4: Preparation of an Extended Release Formulation Containing Edonentan

[0116] A concentrated Edonentan dispersion is made by combining Edonentan with water, Vitamin E-TPGS and y-cyclodextrin. These ingredients are mixed to disperse the Edonentan, and then autoclaved. Sodium hyaluronate may be purchased as a sterile powder or sterilized by filtering a dilute solution followed by lyophilization to yield a sterile powder. The sterile sodium hyaluronate is dissolved in water to make an aqueous concentrate. The concentrated Edonentan dispersion is mixed and added as a slurry to the sodium hyaluronate concentrate. Water is added in sufficient quaintly (q.s., as much as suffices, in this case as much as is required to prepare the homogenous mixture, dispersion, gel or suspension) and the mixture is mixed until homogenous. Examples of these compositions are provided in Table 4 below:Table 4. Compositions of extended release formulation containing Edonentan

[0117] These exemplary compositions contain a sufficient concentration of high molecular weight (z.e. polymeric) sodium hyaluronate so as to form a gelatinous plug or drug depot upon intravitreal injection into a human eye. Preferably the average molecular weight of the hyaluronate used is less than 2 million, and more preferably the average molecular weight of the hyaluronate used is between about 1.3 million and 1.6 million. The Edonentan particles are, in effect, trapped or held within this viscous plug of hyaluronate, so that undesirable pluming does not occur upon intravitreal injection of the formulation. Thus, the risk of drug 29I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOparticles disadvantageously settling directly on the retinal tissue is substantially reduced, for example, relative to using a composition with a water-like viscosity, such as Kenalog® 40. Since sodium hyaluronate solutions are subject to dramatic shear thinning, these formulations are easily injected via 25 gauge, 27 gauge or even 30 gauge needles.EXAMPLE 5: Preparation of a Topical Edonentan Formulation

[0118] A topical Edonentan formulation can be prepared following a known method (e.g., WO 2016156639 Al). More specifically, 20 g of Cremophor® RH40 is dissolved in 75 mL of deionized water by magnetic stirring, which is allowed to stir until completely dissolved. Then 1.5 g of trometamol is added to the resulting solution and stirred for 15 minutes, achieving complete dissolution. 0.5 g of Edonentan is added and allowed to stir for 15 minutes, ensuring complete dissolution. Then 2 g of glycine and 1 g of boric acid are added and allowed to stir until completely dissolved. The resulting solution is added 100 mL deionized water in sufficient quantity. The final solution is filtered with filter paper, and a clear, colorless solution with a pH of 8.06 is obtained. The solution in dropper bottles eyedrop with a volume of 5 mL is packed.EXAMPLE 6: Topical Ophthalmic Solution Nanoparticles Containing Edonentan

[0119] Nanoparticles were prepared by solvent evaporation technique. A solution of 120 mg of 50:50 PLGA in 60 mL of ethyl acetate was prepared. To this solution it was incorporated under turboagitation an aqueous solution of 50 ml of water with 12 mg of Edonentan and 0.5 mg of polyvinyl alcohol. The resulting mixture was left under continuous agitation and under vacuum for 2 hours. Then the resulting preparation was ultra-centrifuged and washed with water three times to remove the nanoparticles from the medium. The nanoparticles thus obtained were dried in a vacuum oven and after evaluation, dispersed in an isotonic aqueous solution enough for a concentration of 5 mg / 1 mL of Edonentan.EXAMPLE 7: Combination therapy with Edonentan and a VEGF / angiopoietin 2 inhibitor results in improved vision and central subfield thickness

[0120] A diabetic retinopathy patient was enrolled in a six-month Phase 2a study. At screening, the patient exhibited no signs of macular edema and received a sustained release 30I PTS / 2OO316166.1Attorney Docket No.: PFT-020WObioerodible implant (Edonentan Implant 1) containing approximately 200 pg of Edonentan, delivered to the intravitreal space via an applicator. Edonentan Implant 1 releases Edonentan over a period of six months.

[0121] At screening, administration of Edonentan Implant 1, and at various timepoints thereafter, visual acuity and central subfield thickness (CST) were assessed. The patient was also examined for retinal neovascularization.

[0122] The patient exhibited macular edema at the time of the 3-month visit. A single dose of 6 mg of Vabysmo® (faricimab-svoa; a VEGF / angiopoietin 2 antibody) was administered at that time by intravitreal injection. Visual acuity and CST continued to be assessed at subsequent visits.Materials and MethodsBest Corrected Visual Acuity test.

[0123] Best Corrected Visual Acuity was measured using an Early Treatment of Diabetic Retinopathy Study (ETDRS) chart (5 letters per row, with standardized spacing between letters and rows, having a total of 14 rows (70 letters). The letter score is calculated when 20 or more letters are read correctly at 4.0 meters. The visual acuity letter score is equal to the total number of letters read correctly at 4.0 meters plus 30 (allowing a maximum score of 100). If fewer than 20 letters are read correctly at 4.0 meters, the visual acuity letter score is equal to the total number of letters read correctly at 4.0 meters (number of letters recorded on line 1.0), plus the total number of letters in the first six lines read correctly at 1.0 meter.Optical coherence tomography (OCT) measurements for central subfield thickness

[0124] The patient underwent detailed fundus evaluation using stereoscopic slit lamp biomicroscopy and indirect ophthalmoscopy. Digital fundus photography and fluorescein angiography were performed. The patient underwent macular thickness analysis using the macular cube 512 x 128 feature of SD-OCT. Multiple OCT-derived values were generated for each scan with values corresponding to the average thickness of a macular field. The central subfield thickness (CST) corresponds to the 1 mm diameter center of the fovea and is surrounded by concentric bands of 3 and 6 mm.31I PTS / 2OO316166.1Attorney Docket No.: PFT-020WOFrequency Doubling Perimetry (FDP) measurements.

[0125] The Matrix perimeter (Carl Zeiss Meditec, Dublin, California, USA) was used to measure the FDP 30-2 visual field. The Matrix stimulus is a 0.25 cycles per degree sinusoidal grating which is phase reversed at 18 Hz. The grating appears to have twice as many alternating light and dark bars than are actually present. The minimum contrast threshold of the 5.08-diameter stimulus is measured at each of the 55 test locations. The frequency doubling illusion on which FDP is based is thought to arise in the magnocellular and be ganglion cell dependent.Results.

[0126] The patient appeared to exhibit an increase in visual acuity after administration of Edonentan Implant 1, up until the three-month visit, when visual acuity abruptly dropped. (FIG. 1A). At the same visit, the patient exhibited signs of macular edema (as evidenced by a dramatic increase in central subfield thickness (CST) measured at that visit; FIG. IB) and retinal neovascularization (observed by fluorescein angiography and fundus photography) and was administered Vabysmo®.

[0127] At the four-month visit (4 weeks after administration of Vabysmo®, the patient’s macular edema, decreased visual acuity (FIG. 1A), and retinal neovascularization were all resolved, with the BCVA score increasing 18 letters from the score at the 3-month visit (FIG.1A) and a 152 pm improvement in the CST. For comparison, at 4 weeks after administration of Vabysmo® as a monotherapy, subjects historically exhibit an average of a 6-letter increase on the BCVA score and a 120 pm improvement in the CST.

[0128] In the patient in this study, resolution of symptoms appeared sustained through the six-month timepoint.

[0129] The above-mentioned results are consistent with the analyses of peripheral vision as assessed by FDP, which showed that the patient’s vision was improving before they experienced macular edema, was transiently worse during the time they experienced macular edema, and then improved again after Vabysmo treatment. (FIG. 2)

[0130] As the Vabysmo® was administered at a time when the patient was still receiving Edonentan, these results demonstrate that a combination of Edonentan and an anti-VEGF therapy resulted in an improvement in at least visual acuity and CST at a level greater than 32I PTS / 2OO316166.1Attorney Docket No.: PFT-020WGthat observed with anti-VEGF therapy alone. Of note, no other therapy to date, include anti-VEGF therapy, is known to improve peripheral vision in diabetic retinopathy or macular edema patients. Typically, peripheral vision is lost in all diabetic retinopathy patients over time.OTHER EMBODIMENTS

[0131] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features set forth herein.33I PTS / 2OO316166.1

Claims

Attorney Docket No.: PFT-020WGCLAIMS1. A method for ameliorating macular edema in a subject in need thereof, the method comprising a step of:co-administering to the subject (1) a sustained release bio-erodible device having a total amount of between about 150 pg and about 650 pg of a crystalline form of Edonentan, by implanting the bio-erodible device into the vitreous cavity of the subject’s eye and (2) a bispecific VEGF / angiopoietin 2 antibody, wherein the subject exhibits:a) improved visual acuity as evidenced by an increase relative to a reference level in the Best Corrected Visual Acuity (BCVA) score by at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, or at least 18 Early Treatment Diabetic Retinopathy (ETDRS) letters, at four weeks after administration of the bispecific VEGF / angiopoietin 2 antibody,b) a reduction relative to a reference level in at least 20 pm, at least 40 pm, at least 60 pm, at least 80 pm, at least 100 pm, at least 120 pm, at least 140 pm, or at least 150 pm in the Central Subfield Thickness (CST) as determined by optical coherence tomography, at four weeks after administration of the bispecific VEGF / angiopoietin 2 antibody, orc) a visual field mean deviation (VF MD) slope of at least 0.50 dB / year, at least 0.75 dB / year, at least 1.00 dB / year, at least 1.25 dB / year, at least 1.50 dB / year, at least 1.75 dB / year, at least 2.00 dB / year, at least 2.25 dB / year, at least 2.50 dB / year as determined by a Frequency Doubling Perimetry (FDP) over a period of at least or about three months, at least or about four months, at least or about five months, or at least or about six months after implanting,wherein the reference level is a level determined by the same assay at a timepoint when the subject is determined to be experiencing macular edema.

2. A method for ameliorating macular edema in a subject in need thereof, the method comprising co-administering to the subject (1) a first composition comprising Edonentan, a pharmaceutically acceptable salt thereof, a crystalline form thereof, or an amorphous form thereof, and (2) a second composition comprising a VEGF inhibitor.34I PTS / 2OO316166.1Attorney Docket No.: PFT-020WO3. In a method of treating a subject in need thereof with a VEGF inhibitor, the improvement comprising co-administering a first composition comprising Edonentan, a pharmaceutically acceptable salt thereof, a crystalline form thereof, or an amorphous form thereof.

4. The method of claim 2 or 3, wherein the subject is a mammal.

5. The method of claim 4, wherein the subject is a human.

6. The method of any one of claims 1-5, wherein the subject is suffering from, at risk of developing, or diagnosed with an eye disease or disorder.

7. The method of claim 6, wherein the eye disease or disorder is selected from the group consisting of age-related macular degeneration, diabetic retinopathy, eye tumor, glaucoma, retinal vein occlusion, retinitis pigmentosa, retinoschisis, uveitis, and vitreomacular traction.

8. The method of claim 7, wherein the eye disease or disorder is diabetic retinopathy.

9. The method of any one of claims 1-8, wherein the subject has undergone an eye surgery or has experienced an injury to the eye.

10. The method of any one of claims 2-9, wherein the subject exhibits a reduction in the central subfield thickness after co-administration.

11. The method of claim 10, wherein the reduction comprises a reduction of least 125 m at four weeks after co-administration.

12. The method of any one of claims 2-11, wherein the subject exhibits an improvement in visual acuity after co-administration.

13. The method of claim 12, wherein the improvement in visual acuity comprises an increase of at least 8 Early Treatment Diabetic Retinopathy (ETDRS) letters on a Best Corrected Visual Acuity test at four weeks after co-administration.

14. The method of any one of claims 2-13, wherein the subject exhibits reduced retinal neovascularization.352OO316166.1Attorney Docket No.: PFT-020WG15. The method of any one of claims 2-14, wherein co-administration comprises intravitreal administration of the first composition.

16. The method of claim 15, wherein intravitreal administration comprises implanting a sustained release, bio-erodible device which comprises the first composition in the vitreous cavity of the eye.

17. The method of claim 1 or 16, wherein the bio-erodible device comprises a poly(lactic- co-glycolic acid) (PLGA) polymer.

18. The method of claim 1, 16, or 17, wherein the bio-erodible device comprises a total amount of about 200 ptg, about 400 ptg, or about 500 ptg of Edonentan.

19. The method of any one of claims 1-18, wherein the administration comprises administering the composition at a regular interval.

20. The method of claim 19, wherein the regular interval is at least three months.

21. The method of claim 19, wherein the regular interval is about four months.

22. The method of claim 19, wherein the regular interval is about six months.

23. The method of any one of claims 2-22, wherein the VEGF inhibitor comprises a VEGF antibody.

24. The method of claim 23, wherein the VEGF antibody comprises a bispecific VEGF / angiopoietin 2 antibody.

25. The method of claim 24, wherein the bispecific VEGF / angiopoietin 2 antibody is faricimab.

26. The method of claim 23, wherein the VEGF antibody comprises a monospecific VEGF antibody.

27. The method of any one of claims 2-22, wherein the VEGF inhibitor comprises a fusion protein comprising a VEGF-R receptor.

28. The method of any one of claims 2-22, wherein the VEGF inhibitor comprises a small molecule.36OO316166.1Attorney Docket No.: PFT-020WG29. The method of any one of claims 1-28, wherein the first composition is administered separately from the second composition or the VEGF inhibitor or bispecific VEGF / angiopoietin 2 antibody.

30. The method of claim 29, wherein the first composition is administered before the second composition, VEGF inhibitor, or bispecific VEGF / angiopoietin 2 antibody is administered.

31. The method of claim 29, wherein the second composition, VEGF inhibitor, or bispecific VEGF / angiopoietin 2 antibody is administered before the first composition is administered.

32. The method of any one of claims 1-28, wherein the first composition and the second composition, VEGF inhibitor, or bispecific VEGF / angiopoietin 2 antibody are administered at the same time.

33. A composition comprising (1) Edonentan, a pharmaceutically acceptable salt thereof, a crystalline form thereof, or an amorphous form thereof, and (2) a VEGF inhibitor, wherein the Edonentan and VEGF inhibitors are present in amounts that are therapeutically effective in combination.37OO316166.1

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