Treatment of ocular disorders

Abstract

The present invention relates to treatment of ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels and particularly, although not exclusively, to the treatment of such ocular disorders with a LINK_TSG6 polypeptide.

Description

[0001] 8849846

[0002] Treatment of Ocular Disorders

[0003] This application claims priority from GB2416087.1 filed 31 October 2024, the contents and elements of which are herein incorporated by reference for all purposes.

[0004] Technical Field

[0005] The present invention relates to the treatment of ocular disorders and particularly, although not exclusively, to the treatment of ocular disorders with a Link_TSG6 polypeptide.

[0006] Background

[0007] Leakage of fluid and / or blood from abnormal or damaged blood vessels (e.g. caused by choroidal neovascularisation (CNV)) can result in macula oedema and visual impairment. The primary treatment for ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is intravitreal injection of anti-VEGF agents. However intravitreal injections are associated with low compliance and a risk of complications that increases with the frequence of injection (Herrero-Vanrell and Refojo, Adv Drug Deliv Rev. (2001) 52(1 ):5-16).

[0008] Tumour necrosis factor (TNF)-stimulated gene 6 (TSG-6) is the ~35 kDa secreted protein product of TNF- stimulated gene-6, expressed in response to inflammatory mediators and growth factors. The sites and contexts of TSG-6 expression, its structure and ligand-binding properties, and how these together underpin its diverse biology and therapeutic potential at a molecular level are reviewed in Day & Milner (Matrix Biology (2019) 78-79, 60-83). For example, Leali et al. (Arterioscler Thromb Vase Biol. (2012) 32(3):696-703) showed that TSG-6 has a pro angiogenic function. The Link module of human TSG-6 (Link_TSG6) is described in US2015 / 0057229 for use in inhibiting cartilage degradation and in WO2021 / 013452 for treating dry eye disease.

[0009] Full-length TSG-6 has also been implicated for use in treating chemical and mechanical injury to the eye. For example, injection of recombinant TSG-6 into the anterior chamber of the eye has been shown to reduce inflammatory damage to the cornea after chemical and mechanical injury (Oh et al., (2012) 107(39):16875-80). Furthermore, Chan et al. (Invest Ophthalmol Vis Sci. (2012) 53(14):1228) and Tuo et al. (J Neuroinflammation (2012) 9:59) found that in a mouse model of AMD (Cc / 2'1' ICxScr1' mice), intravitreal injection of recombinant TSG-6 resulted in a slower progression of retinal lesions. Kim et al. (Sci Rep (2015) 5:11872) found that intravitreal TSG-6 inhibits CNV development and VEGF expression in a rat model of CNV induced by laser photocoagulation.

[0010] Like anti-VEGF agents, previous attempts to utilise TSG-6 in the eye suffer from the disadvantages associated with intravitreal injection into the eye. They are also limited by the concentration of recombinant TSG-6 that can be used. For example, the authors of Kim et al. 2015 refer to a maximal dose of recombinant TSG-6 according to its solubility limit. 8849846

[0011] The present invention has been devised in light of the above considerations.

[0012] Summary of the Invention

[0013] The present disclosure relates to Link_TSG6 polypeptide for use in the treatment of ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels.

[0014] As shown in the examples, Link_TSG6 treatment resulted in fewer leaky lesions in a CNV model.

[0015] In a first aspect, the present disclosure provides a Link_TSG6 polypeptide for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels.

[0016] In some aspects, the ocular disorder is wet AMD, macular oedema secondary to retinal vein occlusion (branch RVO or central RVO), diabetic macular oedema (DME) or myopic choroidal neovascularisation (myopic CNV).

[0017] In some aspects, the treatment comprises subcutaneous administration of Link_TSG6. In other words, Link_TSG6 is administered subcutaneously. Optionally, the Link_TSG6 is administered subcutaneously into the abdomen, thigh or upper arm.

[0018] The Link_TSG6 may be administered subcutaneously at a frequency of 1x per week, 1x per month, 1x per 8 weeks, or 1x per 12 weeks. In some aspects, the Link_TSG6 is administered subcutaneously at a dose of 0.5 mg to 200 mg of Link_TSG6 per dose.

[0019] In some aspects, the treatment comprises intravitreal administration of Link_TSG6. In other words, Link_TSG6 is administered intravitreally. Optionally, the intravitreal administration at a frequency of 1x per month.

[0020] In some aspects, the Link_TSG6 is administered intravitreally at a dose of about 0.05 mg to 10 mg Link_TSG6 per dose.

[0021] In some aspects, the treatment comprises topical administration of Link_TSG6 to the eye. In other words, the Link_TSG6 is administered by topical administration to the eye (e.g. eye drops). Optionally, the topical administration is at a frequency of 2x a day.

[0022] In some aspects, the Link_TSG6 is administered topically at a dose of 100 pg to 500 pg per dose.

[0023] In some aspects, the Link_TSG6 polypeptide comprises, consists, or consists essentially of (i) the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9, or (ii) an amino acid sequence having at least 80% 8849846 identity to the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9. In some aspects, the Link_TSG6 polypeptide comprises, consists, or consists essentially of (i) the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9, or (ii) an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9.

[0024] In some aspects, the Link_TSG6 polypeptide is administered in combination with an anti-VEGF therapy. Anti-VEGF agents may be used to provide complementary effects on inhibiting the growth of new blood vessels and reduce vascular permeability.

[0025] In a yet further aspect, the present disclosure provides a Link_TSG6 polypeptide for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein in a laser-induced choroidal neovascularisation model, a reduction in the percentage of leaky lesions of at least 25% is observed following treatment with the Link_TSG6 polypeptide compared to a control treatment.

[0026] The invention includes the combination of the aspects and preferred features described except where such a combination is clearly impermissible or expressly avoided.

[0027] Detailed Description of the Invention

[0028] Aspects and embodiments of the present invention will now be discussed with reference to the accompanying figures. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this text are incorporated herein by reference.

[0029] Ocular Disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels

[0030] The present disclosure relates to the treatment or prevention of ocular disorders, and specially to ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. Disclosed herein is a Link_TSG6 polypeptide for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. That is to say, the ocular disorder may be characterised by leakage of fluids other than blood, leakage of blood, or leakage of both blood and fluids other than blood. The leakage of fluid may be into the retina. That is to say, in methods and uses disclosed herein the ocular disorder may be characterised by leakage of fluid and / or blood into the retina from abnormal or damaged blood vessels.

[0031] The abnormal or damaged blood vessels described herein relate to blood vessels associated with the eye. In some embodiments, the abnormal or damaged blood vessels are retinal blood vessels. In some embodiments, the abnormal or damaged blood vessels originate in the choroid layer of the eye. In some embodiments, the abnormal or damaged blood vessels penetrate the Bruch’s membrane. 8849846

[0032] Leakage of fluid and / or blood from abnormal or damaged blood vessels can result in visual impairment. The visual impairment as described herein may include blurry vision, blurring or distortion of the central vision (which may result in difficulty recognizing familiar faces), floaters (dark spots or lines in central vision), complete loss of central vision, and washed-out colours. Accordingly, also disclosed herein is a Link_TSG6 polypeptide for use in the treatment or prevention of visual impairment resulting from leakage of fluid and / or blood from abnormal or damaged blood vessels. The visual impairment may be blurry vision, blurring or distortion of the central vision (central vision loss), floaters, complete loss of central vision, and / or washed-out colours.

[0033] Leakage of fluid and / or blood from abnormal or damaged blood vessels can be associated with, or caused by, choroidal neovascularisation (CNV). CNV is the growth of new blood vessels that originate in the choroid layer (a vessel-containing layer under the retina) of the eye. These new, abnormal blood vessels can grow through the retinal pigment epithelium (-RPE) into the subretinal space. The RPE is a single cell thick epithelial sheet sitting on top of the Bruch’s membrane. Together the RPE and Bruch’s membrane form the outer blood-retinal barrier (OBRB). The new, leaky vessels often penetrate the OBRB and ingress into the neurosensory retina (retina), resulting in fluid in the subretinal space. The new vessels may also be associated with haemorrhages and hypoxia.

[0034] Leakage of fluid and / or blood from abnormal or damaged blood vessels can result in macular oedema. Macular oedema (also interchangeably called macular oedema) is swelling caused by fluid build-up in the macula. In other words, there is an accumulation of intraretinal or subretinal fluid. The macula is part of the retina at the back of the eye which is responsible for central vision and colour vision. In some embodiments, an individual having ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, has macula oedema. In some embodiments a Link_TSG6 polypeptide is used in the treatment, or prevention of, macular oedema resulting from growth of abnormal blood vessels.

[0035] The Amsler grid is an eye test that can help monitor a person's central visual field and detect early signs of macular oedema. The Amsler grid is a simple square containing a grid of horizontal and vertical lines with a dot in the middle. For an individual experiencing an ocular disorder characterised by a leakage of fluid and / or blood from abnormal or damaged blood vessels, an Amsler grid may appear to have wavy lines or blank spots. For an individual with macular oedema, an individual with CNV, or an individual with macular oedema and CNV, an Amsler grid may appear to have wavy lines or blank spots.

[0036] Ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels (and any resulting macular oedema) can be identified via testable criteria falling within the common general knowledge of the skilled person. In such ocular disorders, visual impairment (e.g. complete or partial central vision loss as identified by a vision test) is identified, in combination with one or more of the following criteria:

[0037] (i) Fluid or bleeding in the retina is observed in an eye examination (e.g. a dilated eye exam), 8849846

[0038] (ii) Blood vessels that are actively leaking within the macula are identified using Fluorescein or Indocyanine green (ICG) angiography,

[0039] (iii) Swelling (oedema) in the macula is identified by using optical coherence tomography (OCT) to measure retinal thickness.

[0040] It will be appreciated that (i), (ii) and (iii) above are all methods known to the skilled person that can be used to detect leakage of fluid and / or blood from abnormal or damaged blood vessels.

[0041] A patient who has been selected as having, or having susceptibility to, an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels may be diagnosed using any of the methods described above. In some embodiments, methods disclosed herein relate to the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels in a subject who has been diagnosed using any of the methods described above, either alone or in combination.

[0042] In some embodiments, methods disclosed herein relate to the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels in a subject who has been diagnosed with macular oedema and / or CNV. For example, by identifying macular oedema, identifying CNV or identifying both macular oedema and CNV, via the methods described above.

[0043] Leakage of fluid and / or blood from abnormal or damaged blood vessels can occur in several retinal diseases, including wet AMD, retinal vein occlusion, diabetic macular oedema and myopic choroidal neovascularisation. In some embodiments, the ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is wet AMD. In some embodiments, the ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is macular oedema secondary to retinal vein occlusion (branch RVO or central RVO). In some embodiments, the ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is diabetic macular oedema (DME). In some embodiments, the ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is myopic choroidal neovascularisation (myopic CNV).

[0044] The mouse laser-induced choroidal neovascularisation (LI-CNV) model has significantly contributed to our current understanding of the etiology of CNV (Salas et al., Biomedicines. (2023) 11 (9):2445). LI-CNV is also an established model for wet AMD as described in Shah et al. (J Vis Exp. (2015) 27:e53502). In this model, a targeted laser injury is administered to the retinal pigment epithelium (RPE) and Bruch's membrane, which in turn induces angiogenesis. The angiogenesis and leaky lesions resulting from laser induction are similar in location and overall appearance to the angiogenesis and oedema observed in human patients with CNV. 8849846

[0045] The primary therapeutic agents used for treating ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels are anti- Vascular Endothelial Growth Factor (VEGF) agents. Anti- VEGF agents are used to inhibit growth of new blood vessels and reduce vascular permeability. Anti- VEGF agents include aflibercept (Eylea®), brolucizumab (Beovu®), ranibizumab (Lucentis®), bevacizumab (Avastin®) and faricimab-svoa (VABYSMO®). These anti-VEGF agents are administered by intravitreal injection i.e. direct injection into the vitreous body of the eye.

[0046] Intravitreal injections are unpleasant, associated with low compliance and have a risk of complications that increases with the frequence of injection (Herrero-Vanrell and Refojo, Adv Drug Deliv Rev. (2001) 52(1 ):5-16) . Risks of intravitreal injections include pain (e.g. due to corneal abrasion), increased sensitivity to light, retinal tear / detachment, infection, endophthalmitis, increased pressure in the eye, uveitis and cataracts (due to inadvertently hitting the lens).

[0047] An association between repeated anti-VEGF injection and risk of retinal nerve fiber layer (RNFL) loss has also been implicated by some studies (Shin et al., Invest Ophthalmol Vis Sci. (2016) 57(4): 1798-806).

[0048] However, despite these disadvantages, there are no approved therapies for treating ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels that use subcutaneous delivery in the context of protein-based therapies (see Thomas et al. Br J Pharmacol. (2022) 179(9):1908-1937). A subcutaneous anti-VEGF treatment is in development (see clinical trial NCT05387837).

[0049] Wet AMD

[0050] The present disclosure also relates to the treatment or prevention of wet AMD using a Link_TSG6 polypeptide. Age-related macular degeneration (AMD) is a progressive eye condition which causes vision loss and primarily affects adults over 50. There are two primary types of AMD; Dry AMD (also called atrophic AMD, geographic atrophy or non-neovascular AMD) and wet AMD (also called exudative or neovascular AMD).

[0051] Wet AMD occurs in about 10-15% of cases. Wet AMD is primarily caused by the growth of abnormal blood vessels (CNV) in the back of the eye beneath the macula which leak fluid and blood, leading to macular damage. Wet AMD can develop from dry AMD and often leads to more significant vision impairment.

[0052] In some embodiments, methods disclosed herein result in a reduction in symptoms of wet AMD. Symptoms of wet AMD include visual impairment such as distorted and reduced (i.e. wavy, fuzzy or blurry) vision, a dark, empty area or blind spot in the centre of vision, washed-out colours or complete loss of central vision. These symptoms can result in difficulty recognizing familiar faces, difficulty reading, difficulty watching TV, and an inability to drive, thus having a huge effect on quality of life. For someone 8849846 with wet AMD, an Amsler grid may appear to have wavy lines or blank spots. Accordingly, methods disclosed herein may result in a reduction in visual impairment such as such as distorted and reduced vision, difficulty recognizing familiar faces, a dark, empty area, or blind spot in the centre of vision, washed-out colours or complete loss of central vision.

[0053] The exact cause of wet AMD is not fully understood, but several factors may contribute (i.e. risk factors). Risk factors include age (being 50 or older), genetics (e.g., having family history of AMD and / or the presence of wet AMD risk loci (see e.g. Fritsche et al. Nat Genet. 2016 48(2): 134-43)), smoking, hypertension and cardiovascular disease. In some embodiments, methods disclosed herein relate to the treatment or prevention of Wet AMD in a subject who has been selected based on risk factors for wet AMD.

[0054] The primary therapeutic agents used for treating wet AMD are anti-VEGF agents (such as ranibizumab, aflibercept, or bevacizumab). These agents are administered directly into the vitreous of the eye to inhibit VEGF activity, reducing new vessel growth and leakage, and preserving vision. Other treatment options for wet AMD include photodynamic therapy (PDT) (Newman, Eye. (2016) 30(2):202-10).). This is a procedure to seal leaking blood vessels or destroy abnormal blood vessel growth in wet AMD.

[0055] Additionally, lifestyle modifications such as quitting smoking, eating a healthy diet rich in fruits and vegetables, maintaining a healthy weight, and protecting eyes from UV light may be beneficial.

[0056] Macular oedema secondary to retinal vein occlusion

[0057] The present disclosure also relates to the treatment or prevention of macular oedema secondary to retinal vein occlusion using a Link_TSG6 polypeptide. Retinal vein occlusion (RVO) is a partial or total blockage in a vein that drains blood from the retina. Blockage of retinal veins increases pressure and causes fluid to leak from retinal blood vessels, leading to macular oedema. Accordingly, RVO is an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels.

[0058] There are two types of RVO; central retinal vein occlusion (CRVO) and branch retinal vein occlusion (BRVO). CRVO occurs when the main retinal vein (central retinal vein) becomes blocked. BRVO is more common and occurs when one of the smaller ‘branch’ veins emptying into the eye’s main vein becomes blocked. In some embodiments, a patient selected for treatment with a Link_TSG6 polypeptide as described herein suffers from CRVO or BRVO. Risk factors for RVO include age, high blood pressure, hardening of the arteries (arteriosclerosis) and diabetes. In some embodiments, methods disclosed herein relate to the treatment or prevention of macular oedema secondary to retinal vein occlusion in a subject who has been selected based on risk factors for RVO.

[0059] In some embodiments, methods disclosed herein result in a reduction in symptoms of RVO as described herein. Symptoms of RVO include distorted and reduced (i.e. wavy, fuzzy or blurry) vision, difficulty recognizing familiar faces, a dark, empty area or blind spot in the centre of vision, floaters, and / or 8849846 complete or temporary loss of central vision. For someone with RVO, an Amsler grid may appear to have wavy lines or blank spots.

[0060] The primary therapeutic agents used for treating RVO are anti-VEGF agents. These are administered as intravitreal injections to help reduce the leakage and swelling in the macula. Drugs like ranibizumab and aflibercept are often used to manage this condition.

[0061] Diabetic macular oedema (DME)

[0062] The present disclosure also relates to the treatment or prevention of diabetic macular oedema (DME) using a Link_TSG6 polypeptide. DME is a complication of diabetes that can cause vision loss or blindness. Chronic high blood sugar levels in diabetes lead to damage and leakage from retinal blood vessels, resulting in fluid accumulation in the macula.

[0063] DME is associated with diabetic retinopathy severity. Diabetic retinopathy is caused by high levels of sugars in the blood, leading to blockages in blood vessels in the retina. The duration of diabetes and a lack of control over blood sugar levels is strongly correlated with prevalence and incidence of DME. Accordingly, diabetic macular oedema and diabetic retinopathy are ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. In some embodiments, a patient selected for treatment with a Link_TSG6 polypeptide as described herein suffers from diabetes, or suffers from diabetic retinopathy. Risk factors for DME include age, a long duration of diabetes, poor control of diabetes, high blood pressure and hyperlipidaemia. In some embodiments, methods disclosed herein relate to the treatment or prevention of DME in a subject who has been selected based on risk factors for DME.

[0064] DME is caused by two main mechanisms; increased vessel permeability (causing leakage of fluid and blood) and increased vessel closure (leading to neovascularisation).

[0065] Broadly speaking, there are two types of diabetic retinopathy. In early diabetic retinopathy (also called non-proliferative diabetic retinopathy (NPDR)), the walls of the blood vessels in the retina weaken. Tiny bulges develop in the blood vessels, which may bleed slightly. As this condition develops from mild NPDR to more severe NPDR (also called pre-proliferative retinopathy), these changes to the blood vessels in the eye become more severe and more widespread, resulting in more significant bleeding into the eye and fluid build-up in the macula (i.e. macular oedema). In advanced diabetic retinopathy (also called proliferative retinopathy), abnormal blood vessels grow in the retina (neovascularisation) due to existing vessels becoming blocked and damaged. These new vessels are weak and bleed easily, resulting in macular oedema.

[0066] In some embodiments, methods disclosed herein result in a reduction in symptoms of DME as described herein. Patients with DME may experience visual impairment including blurry vision, blurring or distortion 8849846 of the central vision (central vision loss), floaters, complete loss of central vision and / or washed-out colours. For someone with DME, an Amsler grid may appear to have wavy lines or blank spots. OCT is very useful in monitoring DME progression and response to treatment.

[0067] The primary therapeutic agents used for treating DME are anti-VEGF agents (e.g. ranibizumab and aflibercept). These are administered as intravitreal injections to reduce vascular permeability and fluid leakage, thereby decreasing macular oedema and improving visual acuity.

[0068] Myopic choroidal neovascularisation (myopic CNV)

[0069] The present disclosure also relates to the treatment or prevention of myopic choroidal neovascularisation (myopic CNV) using a Link_TSG6 polypeptide. Myopia (also called near-sightedness) is an optical condition caused by having a longer than average eyeball. In patients with high myopia, the retina can become thin and prone to certain problems e.g. the development of abnormal blood vessels under the retina, which can leak and cause visual disturbances. Accordingly, myopic CNV is an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels as described herein. In its advanced stage, myopic CNV can appear as a Fuchs’ spot which is a macular scar with pigment clumping and hyperpigmentation in association with retinal atrophy.

[0070] Risk factors for myopic CNV include age, axial length (extent of myopia) and existing CNV in the other eye. In some embodiments, a patient selected for treatment with a Link_TSG6 polypeptide as described herein suffers from high myopia. In some embodiments, methods disclosed herein relate to the treatment or prevention of myopic CNV in a subject who has been selected based on risk factors for myopic CNV.

[0071] In some embodiments, methods disclosed herein result in a reduction in symptoms of myopic CNV as described herein. Patients with myopic CNV may experience visual impairment including blurry vision, blurring or distortion of central vision (central vision loss), sudden vision loss, floaters, complete loss of central vision and / or washed-out colours. For someone with myopic CNV, an Amsler grid may appear to have wavy lines or blank spots. OCT is very useful in monitoring myopic CNV progression and response to treatment.

[0072] The primary therapeutic agents used for treating myopic CNV are anti-VEGF agents (such as ranibizumab and aflibercept). These are administered as intravitreal injections to inhibit the growth of abnormal vessels and reduce leakage, improving vision outcomes.

[0073] TSG-6 (Tumor Necrosis Factor-Stimulated Gene-6)

[0074] Methods disclosed herein relate to a Link_TSG6 polypeptide for use in the treatment or prevention of ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. Link_TSG6 is a short recombinant peptide comprising the Link module of human TSG-6. 8849846

[0075] TSG-6 is a secreted protein composed of two modular domains. TSG-6 is not usually constitutively expressed in adult tissues, rather being induced in response to inflammatory mediators. During inflammation, TSG-6 is an endogenous protector of tissues. Many of the immunomodulatory and tissue- protective effects of by mesenchymal stem / stromal cells (MSCs) are mediated by their secretion of TSG- 6.

[0076] While constitutively expressed in a few tissues, TSG-6 is generally upregulated wherever there is inflammation. For the most part TSG-6 exhibits anti-inflammatory and tissue protective properties but has been implicated as sometimes playing a role in disease pathology, for example, in the lung. This relates to the enzymatic activity of TSG-6, where it catalyses the covalent transfer of heavy chains (HC) from the inter-a-inh ibitor (lai) family of proteoglycans onto the glycosaminoglycan hyaluronan (HA), to form HOHA complexes (see below). Similarly, TSG-6 has also been shown to drive pathology after chemical injury to the eye (see e.g. Verma et al. Ocul Surf. (2024) 32:26-38). While being made by a broad range of cell types, it was the finding that TSG-6 is produced by mesenchymal stem / stromal cells (MSCs) in response to inflammatory signals, and that it mediates many of their immunomodulatory and reparative activities, which has led to a wealth of publications on the therapeutic effects of this intriguing molecule across a wide range of disease models.

[0077] Recombinant full-length TSG-6 protein has been shown to have anti-inflammatory and tissue protective effects in a wide range of disease models, such as atherosclerosis, myocardial infarction, hypertrophic scarring, colitis, autoimmune diabetes, rheumatoid arthritis, traumatic brain injury or acute lung injury. WO2011 / 139357 described the use of adult stem cells / progenitor cells and stem cell proteins for the treatment of eye injuries and diseases. They propose therapy based on the discovery that after a chemical burn to the cornea of a rat, application of MSCs or MSC conditioned medium reduced inflammation and revascularisation. They proposed the use of anti-apoptotic and anti-inflammatory proteins such as STC-1 and TSG-6, which are expressed by mesenchymal stem cells. Corneal surface inflammation was created in rat eyes by ethanol application and mechanical debridement of the corneal and limbal epithelium. Application of recombinant full-length TSG-6 resulted in reduced corneal opacity and neovascularisation as compared to a PBS control, and the authors conclude that proteins produced by MSCs in response to an injury signal can protect the corneal surface from damage by increasing the viability and proliferation of corneal epithelial progenitors and by suppressing inflammation at the corneal surface.

[0078] TSG-6 is a relatively small protein, with a molecular mass of only ~35-38 kDa, being mainly composed of two modular domains (Link and CUB_C). Given TSG-6's size, it has a surprisingly large number of activities, including the modulation of immune and stromal cell function and its contribution to extracellular matrix formation, mechanics and remodelling. It is the ability of TSG-6 to regulate matrix organization, and to control the association of matrix molecules with cell surface receptors and with extracellular signalling factors (e.g. chemokines), that likely underlies its diverse functional repertoire. In this regard, TSG-6 interacts with a large array of ligands, such as glycosaminoglycans (GAGs), proteoglycan (PG) 8849846 core proteins and other matrix components, and binds directly to multiple chemokines and bone morphogenetic proteins (BMPs). One particularly unusual function of TSG-6 is its role as an enzyme that catalyses the covalent modification of the non-sulphated GAG hyaluronan (HA) with so-called heavy chains (HCs) from the inter-a-inh ibitor (lai) family of proteoglycans. This process, mediated by the full- length TSG-6 protein, but not Link_TSG6 polypeptides containing only a fragment of TSG-6, results in the formation of HOHA complexes, and is essential for mammalian ovulation and fertilisation, and also occurs in many other contexts (e.g. inflammation) where HOHAs either confer tissue protection or contribute to pathological processes.

[0079] The sites and contexts of TSG-6 expression, its structure and ligand-binding properties, and how these together underpin its diverse biology and therapeutic potential at a molecular level are reviewed in Day & Milner (Matrix Biology (2019) 78-79, 60-83).

[0080] Link_TSG6

[0081] A Link_TSG6 polypeptide as disclosed herein comprises the Link module of human or mammalian TSG- 6. In some embodiments, the TSG-6 polypeptide comprises or consists essentially of the amino acid sequence according to SEQ ID NO: 2 or SEQ ID NO: 5. The Link module corresponds to residues 37- 128 of SEQ ID NO:s 2 and 5, and is shown in SEQ ID NO: 7.

[0082] Full length TSG-6 is hard to make, insoluble and prone to aggregation. As disclosed herein, these disadvantages are not associated with Link_TSG6, a short recombinant peptide comprising the Link module of human TSG-6. This short polypeptide is easier to make than full length TSG-6, and is highly soluble and stable in solution.

[0083] The Link module is responsible for the hyaluronan (HA) binding activity, chondroitin sulphate binding activity, aggrecan binding activity, inter-a-inhibitor (lai) binding activity, heavy chain (HC) 1 binding activity, HC2 binding activity, HC3 binding activity, bikunin binding activity, versican binding activity, dermatan sulphate binding activity, pentraxin-3 binding activity, thrombospondin-1 binding activity, thrombospondin-2 binding activity, fibronectin binding activity, heparin / heparan sulphate binding activity, RANKL binding activity, bone morphogenetic protein (BMP) -2 binding activity, BMP-4 binding activity, BMP-5 binding activity, BMP-6 binding activity, BMP-7 binding activity, BMP-13 binding activity, BMP-14 binding activity, CXCL4 binding activity, CXCL6 binding activity, CXCL8 binding activity, CXCL11 binding activity, CXCL12 binding activity, CCL2 binding activity, CCL5 binding activity, CCL7 binding activity, CCL19 binding activity, CCL21 binding activity, CCL27 binding activity or fibroblast growth factor-2 (FGF2) binding activity.

[0084] Link_TSG6 may be a fragment of TSG-6 exhibiting one or more of hyaluronan (HA) binding activity, chondroitin sulphate binding activity, aggrecan binding activity, inter-a-inhibitor (lai) binding activity, heavy chain (HC) 1 binding activity, HC2 binding activity, HC3 binding activity, bikunin binding activity, versican 8849846 binding activity, dermatan sulphate binding activity, pentraxin-3 binding activity, thrombospondin-1 binding activity, thrombospondin-2 binding activity, fibronectin binding activity, heparin / heparan sulphate binding activity, RANKL binding activity, bone morphogenetic protein (BMP) -2 binding activity, BMP-4 binding activity, BMP-5 binding activity, BMP-6 binding activity, BMP-7 binding activity, BMP-13 binding activity, BMP-14 binding activity, CXCL4 binding activity, CXCL6 binding activity, CXCL8 binding activity, CXCL11 binding activity, CXCL12 binding activity, CCL2 binding activity, CCL5 binding activity, CCL7 binding activity, CCL19 binding activity, CCL21 binding activity, CCL27 binding activity or fibroblast growth factor- 2 (FGF2) binding activity.

[0085] In some embodiments, the Link_TSG6 polypeptide is a fragment of TSG-6 exhibiting CCL2, CCL5, CXCL6, CXCL8, and / or CXCL12 binding activity. In some preferred embodiments, Link_TSG6 is a fragment of TSG-6 exhibiting CXCL6 binding activity.

[0086] The Link domain of TSG-6 (Link_TSG6) is the region of full-length TSG-6 N-terminal to the CUB_C domain. As such, the Link_TSG6 protein may lack all or part of the CUB_C domain. In preferred aspects, the Link_TSG6 polypeptide may comprise the part of TSG-6 N-terminal to the CUB_C domain and the first 5 amino acids of the CUB_C domain.

[0087] The Link_TSG6 polypeptide may comprise, consist, or consist essentially of (i) the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9, or (ii) an amino acid sequence having at least 80% identity to the amino acid sequence of SEQ ID NO: 7 or 9.

[0088] Link_TSG6 is preferably a polypeptide comprising or consisting of: (i) the amino acid sequence of SEQ ID NO: 7 or 9, or (ii) an amino acid sequence having one of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequence of SEQ ID NO: 7 or 9.

[0089] In some cases, Link_TSG6 is a polypeptide comprising or consisting of: (i) the amino acid sequence of SEQ ID NO: 7 or 9, or (ii) an amino acid sequence up to 100 amino acids in length having one of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequence of SEQ ID NO: 7 or 9.

[0090] In some cases, Link_TSG6 consists of the amino acid sequence of SEQ ID NO:7 and optionally 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19 or 20 additional amino acids at the C-terminus, the N- terminus, or at each of the C-terminus and the N-terminus.

[0091] In some cases, Link_TSG6 consists of the amino acid sequence of SEQ ID NO:9 and optionally 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19 or 20 additional amino acids at the C-terminus, the N- terminus, or at each of the C-terminus and the N-terminus. 8849846

[0092] The Link_TSG6 polypeptide may consist of, or consist essentially of, the sequence shown in SEQ ID NO: 7.

[0093] Accordingly, the Link_TSG6 polypeptide may comprise:

[0094] (a) the amino acid sequence of SEQ ID NO: 7;

[0095] (b) a variant thereof having at least 50% identity to the amino acid sequence of SEQ ID NO: 7 and having the ability to reduce the percentage of leaky lesions in a mouse laser-induced CNV model by at least 25%; or

[0096] (c) a fragment of either (a) or (b) having CXCL4 binding activity, CXCL6 binding activity, CXCL8 binding activity, CXCL11 binding activity, CXCL12 binding activity, CCL2 binding activity, CCL5 binding activity, CCL7 binding activity, CCL19 binding activity, CCL21 binding activity or CCL27 binding activity or FGF2 binding activity.

[0097] SEQ ID NO: 9 shows a recombinant polypeptide which includes the Link module of TSG-6 (Link_TSG6). The Link_TSG6 polypeptide preferably consists of, or consists essentially of, the sequence shown in SEQ ID NO: 9.

[0098] Accordingly, the TSG-6 polypeptide used in the invention preferably comprises:

[0099] (a) the amino acid sequence of SEQ ID NO: 9;

[0100] (b) a variant thereof having at least 50% identity to the amino acid sequence of SEQ ID NO: 9 and having the ability to reduce the percentage of leaky lesions in a mouse laser-induced CNV model by at least 25%; or

[0101] (c) a fragment of either (a) or (b) having CXCL4 binding activity, CXCL6 binding activity, CXCL8 binding activity, CXCL11 binding activity, CXCL12 binding activity, CCL2 binding activity, CCL5 binding activity, CCL7 binding activity, CCL19 binding activity, CCL21 binding activity or CCL27 binding activity or FGF2 binding activity.

[0102] Amino acid identity may be calculated using any suitable algorithm. For example, the UWGCG Package provides the BESTFIT program which can be used to calculate homology (for example used on its default settings) (Devereux et al. (1984) Nucleic Acids Research 12, 387-395). The PILEUP and BLAST algorithms can be used to calculate homology or line up sequences (such as identifying equivalent or corresponding sequences (typically on their default settings), for example as described in Altschul (1993) J. Mol. Evol. 36, 290-300; Altschul et al. (1990) J. Mol. Biol. 215, 403-10.

[0103] Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / ). The BLAST algorithm performs a statistical analysis of the similarity between two sequences; see e.g., Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90, 5873-5787. One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two polynucleotide or amino acid sequences would occur by chance. For example, a sequence is considered 8849846 similar to another sequence if the smallest sum probability in comparison of the first sequence to the second sequence is less than about 1 , preferably less than about 0.1 , more preferably less than about 0.01 , and most preferably less than about 0.001 .

[0104] The variant sequences typically differ by at least 1 , 2, 5, 10, 20, 30, 50 or more mutations (which can be substitutions, deletions or insertions of amino acids). For example, from 1 to 50, 2 to 30, 3 to 20 or 5 to 10 amino acid substitutions, deletions or insertions can be made. The modified polypeptide may generally retain the ability to reduce the percentage of leaky lesions in a mouse laser-induced CNV model by at least 25%, or to retain CXCL4 binding activity, CXCL6 binding activity, CXCL8 binding activity, CXCL11 binding activity, CXCL12 binding activity, CCL2 binding activity, CCL5 binding activity, CCL7 binding activity, CCL19 binding activity, CCL21 binding activity, CCL27 binding activity or fibroblast growth factor- 2 (FGF2) binding activity, preferably in a dose-dependent manner. The substitutions are preferably conservative substitutions, for example according to the following Table. Amino acids in the same block in the second column and preferably in the same line in the third column may be substituted for each other:

[0105] A Link_TSG6 polypeptide used in the invention is typically at least 10, for example at least 15, 20, 25, 30, 40, 50, 60, 70, 80, 90 or more amino acids in length, up to 100 or 150 amino acids in length, as long as it retains the ability to reduce the percentage of leaky lesions in a mouse laser-induced CNV model by at least 25%, or the CXCL4 binding activity, CXCL6 binding activity, CXCL8 binding activity, CXCL11 binding activity, CXCL12 binding activity, CCL2 binding activity, CCL5 binding activity, CCL7 binding activity, CCL19 binding activity, CCL21 binding activity, CCL27 binding activity or fibroblast growth factor- 2 (FGF2) binding activity of TSG-6. Preferably, the polypeptide includes the sequence shown in SEQ ID NO: 7.

[0106] Fragments of the amino acid sequence of SEQ ID NO: 7 may be used in the invention. Such fragments preferably contain the residues Lys-10 and / or Tyr-11 and / or Tyr-58 and / or Phe-69 and / or Tyr-77 of SEQ ID NO: 7. Most preferably, the fragment of SEQ ID NO: 7 contains each of residues Lys-10, Tyr-11 , Tyr- 58, Phe-69 and Tyr-77 of SEQ ID NO: 7.

[0107] Fragments of the amino acid sequence of SEQ ID NO: 9 preferably contain the residues Lys-11 and / or Tyr-12 and / or Tyr-59 and / or Phe-70 and / or Tyr-78 of SEQ ID NO: 9. Most preferably, the fragment of SEQ ID NO: 9 contains each of residues Lys-11 , Tyr-12, Tyr-59, Phe-70 and Tyr-78 of SEQ ID NO: 9. 8849846

[0108] The TSG-6 polypeptides used in the invention may be chemically modified, e.g. post-translationally modified. For example, they may be glycosylated, phosphorylated or comprise modified amino acid residues. They may be modified by the addition of histidine residues to assist their purification or by the addition of a transmembrane sequence to promote insertion into the cell membrane. Such modified polypeptides fall within the scope of the term "polypeptide" used herein.

[0109] SEQ ID NO: 9 is LINK_TSG6, which is a polypeptide that consists of the Link module of TSG-6 (corresponding to SEQ ID NO: 7), and additionally a Glycine (Gly) residue at the N-terminus, and five additional amino acid residues at the C-terminus (Asn-Pro-His-Ala-Lys). The extra amino acids present in SEQ ID NO: 9 do not significantly impact the 3D structure of the polypeptide. The polypeptides encoded by SEQ ID NO: 9 and SEQ ID NO: 7 therefore have similar structure and function.

[0110] Suitable assays for determining the ability of a TSG-6 polypeptide to bind to CXCL4, CXCL6, CXCL8, CXCL11 , CXCL12, CCL2, CCL5, CCL7, CCL19, CCL21 and CCL27 are well-known in the art (see e.g. Dyer et al. (2014) J. Immunol 192, 2177-2185; Dyer et al. (2016) J. Biol. Chem. 291 , 12627-12640; and Day & Milner. (2019) Matrix Biology. 78-79, 60-83).

[0111] TSG-6 polypeptides for use in the invention may be in a substantially isolated form. It will be understood that the polypeptide may be mixed with carriers or diluents which will not interfere with the intended purpose of the polypeptide and still be regarded as substantially isolated. A polypeptide for use in the invention may also be in a substantially purified form, in which case it will generally comprise the polypeptide in a preparation in which more than 50%, e.g. more than 80%, 90%, 95% or 99%, by weight of the polypeptide in the preparation is a polypeptide of the invention.

[0112] Link_TSG6 polypeptides for use in the present invention may be natural or non-naturally occurring polypeptides. Polypeptides may be isolated from any suitable organism that expresses a TSG-6 polypeptide. The TSG-6 polypeptide may be isolated from a human or another suitable mammal, such as primates, rats or mice. Alternatively, TSG-6 polypeptide may be isolated from a fish or an amphibian. Polypeptides for use in the invention may also be prepared as fragments of such isolated polypeptides. Further, the Link_TSG6 polypeptides may also be made synthetically or by recombinant means. For example, a recombinant Link_TSG6 polypeptide may be produced by transfecting cells in culture with an expression vector comprising a nucleotide sequence encoding the polypeptide operably linked to suitable control sequences, culturing the cells, extracting and purifying the Link_TSG6 polypeptide produced by the cells. Methods for the recombinant production of polypeptides are well-known in the art (for example, Sambrook et al., 2001 , Molecular Cloning: a laboratory manual, 3rdedition, Cold Spring Harbor Laboratory Press). Preferably, the Link_TSG6 polypeptide is made in a bacteria, such as E. coll.

[0113] The amino acid sequence of Link_TSG6 polypeptides for use in the invention may be modified to include non-naturally occurring amino acids or to increase the stability of the compound. When the polypeptides 8849846 are produced by synthetic means, such amino acids may be introduced during production. The polypeptides may also be modified following either synthetic or recombinant production.

[0114] Link_TSG6 polypeptides for use in the invention may also be produced using D- amino acids. In such cases the amino acids will be linked in reverse sequence in the C- to N-orientation. This is conventional in the art for producing such polypeptides.

[0115] A number of side chain modifications are known in the art and may be made to the side chains of the Link_TSG6 polypeptides, provided that the polypeptides retain the ability to treat ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels.

[0116] Therapeutic and prophylactic applications

[0117] Disclosed herein are methods of treating or preventing an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels.

[0118] The present disclosure provides a Link_TSG6 polypeptide described herein for use in a method of treating or preventing an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. Also provided is the use of a Link_TSG6 polypeptide described herein in the manufacture of a medicament for treating or preventing an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. Also provided is a method of treating or preventing an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, comprising administering to a subject a therapeutically or prophylactically effective amount of a Link_TSG6 polypeptide described herein.

[0119] The methods disclosed herein may result in reduction of the development or progression of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, alleviation of the symptoms of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels or reduction in the pathology of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. The methods may be effective to prevent progression of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, e.g. to prevent worsening of, or to slow the rate of development of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. In some embodiments, treating an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels means preventing worsening of visual impairment resulting from said ocular disorder. In some embodiments, the methods may lead to an improvement in an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, e.g. a reduction in macular oedema. It will be appreciated that in therapeutic embodiments described herein, Link_TSG6 is the active ingredient ( / .e. is responsible for the therapeutic effect). 8849846

[0120] Prevention may mean that no symptoms of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels emerge, or it may mean that the symptoms of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels develop to a lesser extent than in the absence of treatment.

[0121] In the uses and methods described herein, the ocular disorder may be wet AMD, macular oedema secondary to retinal vein occlusion (branch RVO or central RVO), diabetic macular oedema (DME) or myopic choroidal neovascularisation (myopic CNV).

[0122] The methods may involve the reduction, elimination or reduction in development of one or more signs or symptoms of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. These signs or symptoms may include any of elimination or reduction in development of CNV (e.g. fewer leaky lesions or reduced leaky lesion area), elimination or reduction in development of visual impairment (e.g. blurry vision, blurring or distortion of the central vision (central vision loss), floaters, complete loss of central vision and / or washed-out colours), elimination or reduction in development of swelling (oedema) as measured by retinal thickness, and elimination or reduction in development of distorted vision as measured by an Amsler grid.

[0123] Administration of the Link_TSG6 polypeptide to a patient in need thereof may result in a reduction in CNV. A reduction in CNV may be determined by measuring a reduced leaky area. A reduction in CNV may be determined by a reduced number, or a reduced percentage, of leaky lesions. For example, a reduction in leaky lesions may be measured by fluorescein angiography (FA) or an OCT method, such as spectral domain optical coherence tomography (SD-OCT). Representative images of leaky lesions and non-leaky lesions are shown in Figure 6. In some embodiments is a Link_TSG6 polypeptide for use in a method of treating an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels as disclosed herein, wherein administration of the Link_TSG6 polypeptide to a patient in need thereof results in a reduction in CNV. In some embodiments is a method of treating an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels as disclosed herein, the method comprising administering Link_TSG6 to a patient in need thereof, wherein administration of the Link_TSG6 polypeptide to a patient in need thereof results in a reduction in CNV.

[0124] Administration of the Link_TSG6 polypeptide to a patient in need thereof may result in a reduction in leaky lesions. Leaky lesions are areas of new blood vessel growth (e.g. in response to VEGF) where the vessels are fragile / leaky / fenestrated, allowing egress of fluid and potentially red blood cells into surrounding issue. In some embodiments is a Link_TSG6 polypeptide for use in a method of treating an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels as disclosed herein, wherein a reduction in the percentage of leaky lesions is observed following treatment with the Link_TSG6 polypeptide. 8849846

[0125] As described herein, administration of Link_TSG6 in a laser-induced CNV model of wet AMD reduced the percentage of leaky lesions compared to a control treatment (for example, a vehicle (PBS) control treatment). Accordingly, in some embodiments a reduction in the percentage of leaky lesions of at least 25% is observed following treatment with the Link_TSG6 polypeptide compared to a control treatment in a laser-induced choroidal neovascularisation model. In some embodiments, the reduction in leaky lesions of at least 25% is observed at 4 or 7 days after induction of LI-CNV.

[0126] In some embodiments, a method of treating a patient having an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels comprises:

[0127] (i) evaluating a subject or patient for evidence of, or susceptibility to an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels (e.g. by the methods described herein), or providing a patient that has been so evaluated,

[0128] (ii) selecting a patient having, or having susceptibility to an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels if the patient has visual impairment (e.g. complete or partial central vision loss), and at least one of:

[0129] (a) fluid or bleeding in the retina (e.g. as observed during a dilated eye examination),

[0130] (b) blood vessels that are actively leaking within the macula (e.g. as identified using Fluorescein or Indocyanine green (ICG) angiography), and

[0131] (c) swelling (oedema) in the macula (e.g. as identified by OCT),

[0132] (iii) administering Link_TSG6 to a patient in need thereof, optionally wherein the administration is subcutaneous administration, optionally subcutaneous administration to the abdomen, thigh or upper arm; thereby treating the ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels (e.g. reduced CNV) or reducing symptoms thereof.

[0133] The method may involve repeating steps (i) to (iii).

[0134] In any of the methods and uses disclosed herein, the Link_TSG6 polypeptide may be administered in combination with an anti-VEGF therapy (also called a VEGF inhibitor). In such embodiments, the Link_TSG6 polypeptide and the anti-VEGF therapy are both active agents. In such embodiments, the treatment does not comprise a single molecule comprising an anti-VEGF domain and a Link_TSG6 domain. Put another way, there is no administration of a single molecule that has co-domains (a Link_TSG6 domain and an anti-VEGF domain). In other words, the Link_TSG6 and the anti-VEGF are separate molecules and it will be appreciated that the Link_TSG6 polypeptide as described herein is not linked to another protein or nucleic acid.

[0135] The Link_TSG6 polypeptide may be administered simultaneously with an anti-VEGF therapy (simultaneous co-administration of separate molecules), or the Link_TSG6 polypeptide and the anti- VEGF therapy may be administered sequentially. In some embodiments, the Link_TSG6 polypeptide is administered by subcutaneous administration (e.g. as described herein below) and the anti-VEGF therapy 8849846 is administered by intravitreal injection. In some embodiments, the Link_TSG6 polypeptide is administered by topical administration to the eye (e.g. as described herein below) and the anti-VEGF therapy is administered by intravitreal injection. In some embodiments, the Link_TSG6 polypeptide and the anti-VEGF therapy are both administered by intravitreal injection (e.g. as described herein below).

[0136] In some embodiments, the treatment does not comprise an anti-VEGF treatment. In some such embodiments, the treatment does not comprise administering an additional protein that binds VEGF, e.g. an anti-VEGF antibody. Link_TSG6 binds directly to VEGF, therefore it will be appreciated that the phrase “an additional protein that binds VEGF” refers to a protein which is not the Link_TSG6 polypeptide. In other words, in some embodiments a pharmaceutical composition comprising a Link_TSG6 polypeptide as described herein does not comprise an additional protein that binds VEGF.

[0137] The Link_TSG6 polypeptide as described herein is the active agent and is not linked to another protein or nucleic acid that has a therapeutic effect. In other words, the treatments and methods described herein do not comprise administering a single molecule having (i) a LINK_TSG6 domain, and (ii) a protein or nucleic acid domain (e.g. a protein that binds VEGF, C5, Factor P (properdin), Factor D, EPO, EPOR, IL- 1 p, IL-17 A, IL-10, TNFa, or FGFR2). In some embodiments, the treatment does not comprise administering an additional protein that binds VEGF, VEGFR, C5, Factor P (properdin), Factor D, EPO, EPOR, IL-1 p, IL-17A, IL-10, TNFa, Ang-2, PDGF, FGF-2, FGFR1 or FGFR2. It will be appreciated that the phrase “an additional protein” refers to a protein which is not the Link_TSG6 polypeptide. In some such embodiments, the treatment does not comprise administering an additional protein that binds VEGF, C5, Factor P (properdin), Factor D, EPO, EPOR, IL-1 p, IL-17A, IL-10, TNFa, or FGFR2. In other words, in some embodiments a pharmaceutical composition comprising a Link_TSG6 polypeptide as described herein does not comprise an additional protein that binds VEGF, VEGFR, C5, Factor P (properdin), Factor D, EPO, EPOR, IL-1 , IL-17A, IL-10, TNFa, Ang-2, PDGF, FGF-2, FGFR1 or FGFR2. In some such embodiments a pharmaceutical composition comprising a Link_TSG6 polypeptide as described herein does not comprise an additional protein that binds VEGF, C5, Factor P (properdin), Factor D, EPO, EPOR, IL-1p, IL-17A, IL-10, TNFa, or FGFR2.

[0138] The present disclosure provides a Link_TSG6 polypeptide described herein for use in a method of treating or preventing an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein the treatment does not comprise an anti-VEGF treatment. Also provided is the use of a Link_TSG6 polypeptide described herein in the manufacture of a medicament for treating or preventing an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein the medicament does not comprise an anti-VEGF therapy. Also provided is a method of treating or preventing an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, comprising administering to a subject a therapeutically or prophylactically effective amount of a Link_TSG6 polypeptide described herein, wherein the treatment does not comprise an anti-VEGF treatment. 8849846

[0139] Subjects

[0140] A subject in accordance with the various aspects of the present disclosure may be any animal or human. Therapeutic and prophylactic applications may be in human or animals (veterinary use).

[0141] The subject to be administered with Link_TSG6 (e.g. in accordance with therapeutic or prophylactic intervention) may be a subject in need of such intervention. Therapeutic uses may be in human or animals (veterinary use). The subject is preferably mammalian, more preferably human. The subject may be a non-human mammal, but is more preferably human. The subject may be male or female. The subject may be a patient.

[0142] In accordance with the methods of the invention, the methods may additionally comprise the step of selecting a subject for treatment with a therapeutically effective amount of a polypeptide comprising or consisting of Link_TSG6. Patients may be selected for treatment who exhibit evidence of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. The methods may comprise evaluating a subject or patient for evidence of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. For example, evaluating a subject or patient for evidence of wet AMD, retinal vein occlusion (RVO), diabetic macular oedema (DME) and / or myopic choroidal neovascularisation (myopic CNV). The methods may comprise evaluating a patient for evidence of macular oedema. For example, evaluating a patient suffering from wet AMD, retinal vein occlusion, DME or myopic CNV for evidence of macular oedema.

[0143] In some embodiments, the methods may comprise evaluating a subject or patient for evidence of, or susceptibility to an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, such as:

[0144] (i) visual impairment (e.g. complete or partial central vision loss),

[0145] (ii) fluid or bleeding in the retina (e.g. as observed during a dilated eye exam),

[0146] (iii) blood vessels that are actively leaking within the macula (e.g. as identified using Fluorescein (e.g. Fundus Fluorescein Angiography (FFA)) or Indocyanine green (ICG) angiography),

[0147] (iv) A CNV lesion of any type ( / .e., predominantly classic, minimally classic, or occult [including polypoidal choroidal vasculopathy and retinal angiomatous proliferation]) that exhibits all of the following characteristics: o A total lesion size (including e.g. blood, atrophy, and neovascularisation) of <9 disc areas on FFA o A CNV component area of >50% of the total lesion size on FFA o Active CNV confirmed on FFA (evidence of leakage) o CNV exudation confirmed using optical coherence tomography (OCT) (presence of fluid)

[0148] (v) swelling (oedema) in the macula (e.g. as identified by using OCT to measure retinal thickness), 8849846

[0149] (vi) Subfoveal CNV or juxtafoveal / extrafoveal CNV with a subfoveal component related to the CNV activity identified by FFA or OCT (where CNV activity is defined as showing evidence of subretinal fluid, subretinal hyperreflective material, or leakage),

[0150] (vii) increased retinal thickness,

[0151] (viii) a family history of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels (e.g. a family history of wet AMD), and / or

[0152] (ix) age (the subject being >50 years old), and / or

[0153] (x) Best corrected visual acuity (BCVA) of 78-24 letters, inclusive (20 / 32-20 / 320 approximate Snellen equivalent), using the ETDRS protocol (Ophthalmology. (1991) 98(5 Suppl):741-56.) and assessed at the initial testing distance of 4 m on day 1 .

[0154] For example, methods may comprise evaluating a subject or patient for an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels via vision tests (e.g. to identify complete or partial central vision loss), an eye examination (e.g. a dilated eye exam), fluorescein or Indocyanine green (ICG) angiography, tonometry (to identify increased pressure in the eye) or by using optical coherence tomography (OCT) to measure retinal thickness and therefore assess the amount of swelling (oedema) in the macular. Methods may comprise evaluating a subject or patient using an Amsler grid. A subject may be selected for treatment because they see wavy lines or blank spots on an Amsler grid.

[0155] A subject may have (e.g. may have been diagnosed with) wet AMD, may be suspected of having wet AMD, or may be at risk of developing / contracting wet AMD. A subject may have wet AMD in both eyes. The patient may have previously been treated for wet AMD, for example, the patient may have previously received an anti-VEGF therapy to treat wet AMD.

[0156] A subject may have visual impairment due to macular oedema secondary to retinal vein occlusion (branch RVO or central RVO) (e.g. may have been diagnosed with retinal vein occlusion, may be suspected of having retinal vein occlusion, or may be at risk of developing / contracting retinal vein occlusion. A subject may have retinal vein occlusion in both eyes. The patient may have previously been treated for retinal vein occlusion, for example, the patient may have previously received an anti-VEGF therapy to treat retinal vein occlusion.

[0157] A subject may have visual impairment due to diabetic macular oedema (DME) (e.g. may have been diagnosed with DME), may be suspected of having DME, or may be at risk of developing / contracting DME. A subject may have diabetes. A subject may have DME in both eyes. The patient may have previously been treated for DME, for example, the patient may have previously received an anti-VEGF therapy to treat DME.

[0158] A subject may have visual impairment due to myopic choroidal neovascularisation (myopic CNV) (e.g. may have been diagnosed with myopic CNV), may be suspected of having myopic CNV, or may be at risk 8849846 of developing / contracting myopic CNV. A subject may have myopia. A subject may have myopic CNV in both eyes. The patient may have previously been treated for myopic CNV, for example, the patient may have previously received an anti-VEGF therapy to treat myopic CNV.

[0159] Routes of Administration

[0160] The methods described herein involve administration of a therapeutically effective amount of a Link_TSG6 polypeptide. Medicaments and pharmaceutical compositions according to aspects of the present invention may be formulated for administration by a number of routes, including but not limited to, subcutaneous, topical, intravitreal, intra-articular, parenteral, intravenous, intra-arterial, intramuscular, oral and nasal. In some embodiments, the Link_TSG6 polypeptide described herein for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels are administered subcutaneously, intravitreally or topically to the surface of the eye.

[0161] Evaluation of a subject for macular oedema may occur any point before, during or after administration of a therapeutically effective amount of Link_TSG6.

[0162] The methods and uses described herein may comprise ongoing monitoring. For example, monitoring evidence of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, such as visual impairment, fluid or bleeding in the retina, macular oedema and / or increased retinal thickness. In some embodiments, subsequent doses may be administered following monitoring.

[0163] Subcutaneous Administration

[0164] In some embodiments, the Link_TSG6 polypeptide described herein for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is administered subcutaneously. In subcutaneous (SC) administration (also called hypodermic injection), the treatment is given into the fat between the skin and the muscle. In other words, a device (e.g. a syringe) penetrates the epidermis and dermis (layers of the skin) and deposits the product into the subcutaneous fat (adipose tissue) residing under the skin and over the muscle.

[0165] In some embodiments, disclosed herein is a method of treating an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, the method comprising subcutaneously administering Link_TSG6 to a patient in need thereof. In some embodiments is provided the use of a Link_TSG6 polypeptide in the manufacture of a medicament for the treatment of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein the medicament is formulated for subcutaneous administration. 8849846

[0166] SC administered drugs can work systemically and do not necessarily need to be administered at the site of treatment. For example, in the case of treating an ocular disorder, SC administration may comprise administration into a site which is not in or near the eye, or a site which is not the face or head. Exemplary sites of SC administration are the abdomen, thigh or upper arm. The active substance can then be absorbed to the systemic blood circulation through either blood capillaries or via the lymphatic system.

[0167] Accordingly, the Link_TSG6 polypeptide described herein may be administered subcutaneously into a site which is not at or near the eye. In some embodiments, the Link_TSG6 polypeptide described herein is administered, or is for administration, subcutaneously into the abdomen, thigh or upper arm.

[0168] The Link_TSG6 polypeptide described herein may be administered subcutaneously at a frequency of one time (once) per week. The week is preferably a calendar week (i.e. one administration on average every 7 days). In some cases, the treatment comprises SC administration of Link_TSG6 less frequently than once per week, such as once every two weeks, once every three weeks, once every 4 weeks, once every 8 weeks or once every 12 weeks. In some preferred cases, SC administration may be one time per month (one time per calendar month, one administration on average every 30 days), once per 2 months (one time per 2 calendar months, or one administration on average every 60 days) or once per 3 months (one time per 3 calendar months, or one administration on average every 90 days). As used herein, the term “average” refers to the mathematical mean.

[0169] In some cases, one SC administration is given on average every 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30 or 31 days. In some embodiments, one SC administration is given on average every 7, 14, 21 , 28, 30, 60 or 90 days. Preferably, SC administration is given on average every 30 days, every 60 days or every 90 days.

[0170] Intravitreal Administration

[0171] In some embodiments, the Link_TSG6 polypeptide described herein for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is administered intravitreally, i.e. by injection into the vitreous body of the eye. In other words, the treatment comprises intravitreal administration (IVT) of Link_TSG6 polypeptide.

[0172] Also disclosed herein is a method of treating an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein the method comprises intravitreal administration of Link_TSG6 to a patient in need thereof. In some embodiments is provided the use of a Link_TSG6 polypeptide in the manufacture of a medicament for the treatment of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein the medicament is formulated for intravitreal administration. 8849846

[0173] The Link_TSG6 polypeptide described herein may be administered intravitreally at a frequency of one time (once) per week. The week is preferably a calendar week (i.e. one administration on average every 7 days). In some cases, the treatment comprises intravitreally administering Link_TSG6 less frequently than once per week, such as once every two weeks, once every three weeks, once per month or once every 8 weeks. In some preferred cases, IVT administration may be one time per month (one time per calendar month, or one administration on average every 30 days) or one time per 2 months (one time per 2 calendar months, or one administration on average every 60 days).

[0174] Topical Administration

[0175] Methods described herein may involve the topical administration of Link_TSG6. Link_TSG6 may be topically administered to the eye (ocular delivery), preferably to the cornea, such as the surface of the cornea. In some cases, the Link_TSG6 is administered as an eye drop. In other words, the treatment comprises topical administration of Link_TSG6 polypeptide (e.g. administration of an eye drop comprising Link_TSG6) to the eye. As used herein, the term “eye drop” refers to a topical formulation for administration (e.g. dropwise administration) to the ocular surface.

[0176] The Link_TSG6 polypeptide may be formulated with, or the treatment may involve co-administration with prednisolone, artificial tears, or any combination thereof.

[0177] Provided herein is a Link_TSG6 polypeptide for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein the Link_TSG6 polypeptide is administered topically. In some embodiments, disclosed herein is a method of treating an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, the method comprising topically administering Link_TSG6 to a patient in need thereof. In some embodiments is provided the use of a Link_TSG6 polypeptide in the manufacture of a medicament for the treatment of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein the medicament is formulated for topical administration to the eye, such as an eye drop.

[0178] In some embodiments, the Link_TSG6 polypeptide described herein for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is administered topically at a frequency of 1x, 2x, 3x or 4x per day. Preferably, administration is 2x per day. In some embodiments, the treatment comprises topical administration of the Link_TSG6 polypeptide more than 4 times per day. In some cases, the treatment comprises administering the Link_TSG6 polypeptide less frequently than once per day, such as once per week, once every two weeks, once every three weeks or once per month (one time per calendar month, or one administration on average every 30 days). 8849846

[0179] Doses

[0180] Administration is preferably in a "therapeutically effective amount", this being sufficient to show benefit to the individual. For example, an amount sufficient to result in a reduction in visual impairment or a reduction in CNV.

[0181] The actual amount administered, and rate and time-course of administration, will depend on the nature and severity of the disease being treated. Prescription of treatment, e.g. decisions on dosage etc, is within the responsibility of general practitioners and other medical doctors, and typically takes account of the disorder to be treated, the condition of the individual patient, the site of delivery, the method of administration and other factors known to practitioners. Examples of the techniques and protocols mentioned above can be found in Remington’s Pharmaceutical Sciences, 20th Edition, 2000, pub. Lippincott, Williams & Wilkins.

[0182] As disclosed herein, Link_TSG6 was effective at reducing the number of leaky lesions when compared to a control when administered subcutaneously, intravitreally or by topical administration to the eye in a mouse model of CNV.

[0183] In certain aspects, and particularly for subcutaneous administration, the dosage may be scaled for human administration based on body surface area comparisons for mouse and human. In particular, see Nair and Jacob (Journal of basic and clinical pharmacy. 2016; 7(2), 27-31), the contents of which is incorporated herein in its entirety. Example doses were calculated using a human weight of 60 kg.

[0184] In some embodiments, the Link_TSG6 polypeptide described herein for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is administered at a dose of about 0.05 mg to 200 mg of Link_TSG6 per dose.

[0185] Thus, in some aspects, the treatment involves administration of 0.05 mg to 200 mg per month, 0.1 mg to 200 mg per month, 0.5 mg to 200 mg per month, 1 mg to 150 mg per month, 2 mg to 140 mg per month, 5 mg to 130 mg per month, 10 mg to 120 mg per month or 20 mg to 110 mg per month. In some aspects, the treatment involves administration of about 0.1 mg per month, about 0.5 mg per month, about 1 mg per month, about 2 mg per month, about 10 mg per month, about 20 mg per month, about 30 mg per month, about 40 mg per month, about 50 mg per month, about 60 mg per month, about 70 mg per month, about 80 mg per month, about 90 mg per month or about 100 mg per month of Link_TSG6.

[0186] Subcutaneous Administration

[0187] Appropriate human dosage values for subcutaneous administration can be calculated from effective mouse dosages based on body surface area (Nair and Jacob, 2016). Accordingly, in some embodiments, the treatment involves subcutaneous administration of 0.5 mg to 200 mg of Link_TSG6 per dose. In other words, Link_TSG6 is administered subcutaneously at a dose of 0.5 mg to 200 mg Link_TSG6 per dose. 8849846

[0188] In some embodiments the treatment involves subcutaneous administration of 1 mg to 50 mg of Link_TSG6 per dose.

[0189] In some embodiments, the treatment involves subcutaneous administration of at least 0.5 mg, at least 1 mg, at least 2 mg, at least 10 mg, at least 20 mg, at least 30 mg, at least 40 mg, at least 50 mg, at least 60 mg, at least 70 mg, at least 80 mg, at least 90 mg, at least 100 mg or more than 100 mg per dose.

[0190] In some embodiments, the treatment involves subcutaneous administration of between 0.5 mg to 200 mg, between 1 mg to 150 mg, between 1 mg to 100 mg, 2 mg to 100 mg, 1 mg to 50 mg, 2 mg to 50 mg, between 10 mg to 50 mg, between 20 mg to 50 mg, between 30 mg to 50 mg, or between 20 mg to 40 mg of Link_TSG6 polypeptide per dose. In some embodiments, a high dose is required, and the treatment involves subcutaneous administration of between 50 mg to 200 mg, 100 mg to 200 mg, 120mg to 200 mg, 140 mg to 200 mg, 160 mg to 200 mg, or 180 mg to 200 mg per dose. In some embodiments, the treatment involves subcutaneous administration of about 0.5 mg, about 1 mg, about 2 mg, about 5 mg, about 10 mg, about 20, about 30, about 40, about 50, about 100, about 150 or about 200 mg Link_TSG6 polypeptide per dose.

[0191] In some embodiments, the treatment involves subcutaneous administration of one dose (e.g. a dose as described hereinabove, e.g. a dose of 0.5 mg to 200 mg) of Link_TSG6 polypeptide every once per week, once every two weeks, once every three weeks, once every 4 weeks, once every 8 weeks or once every 12 weeks, preferably every 8 or 12 weeks. By way of example, in some embodiments, the treatment involves subcutaneous administration of between 1 mg to 50 mg Link_TSG6 polypeptide once per week, once every two weeks, once every three weeks, once every 4 weeks, once every 8 weeks or once every 12 weeks, preferably every 8 or 12 weeks. In some embodiments, the treatment involves subcutaneous administration of at least 0.5 mg, at least 1 mg, at least 2 mg, at least 10 mg, at least 20 mg, at least 30 mg, at least 40 mg, at least 50 mg, at least 60 mg, at least 70 mg, at least 80 mg, at least 90 mg or at least 100 mg of Link_TSG6 polypeptide, once per week, once every two weeks, once every three weeks, once every 4 weeks, once every 8 weeks or once every 12 weeks, preferably every 8 or 12 weeks.

[0192] Intravitreal Administration

[0193] Appropriate human dosage values for intravitreal administration can be calculated from effective mouse dosages using a 1000x multiplier based on the relative vitreous volumes of human and mouse (~5 ml and ~0.005 ml). Accordingly, in some embodiments, the treatment involves intravitreal administration of between 0.05 mg to 10 mg Link_TSG6 polypeptide per dose. In other words, Link_TSG6 is administered intravitreally at a dose of 0.05 mg to 10 mg Link_TSG6 per dose. In some embodiments, the treatment involves intravitreal administration of 0.1 mg to 5 mg of Link_TSG6 per dose. 8849846

[0194] In some embodiments, the treatment involves intravitreal administration of at least 0.05 mg, at least 0.06 mg, at least 0.08 mg, at least 0.1 mg, at least 0.12 mg, at least 0.15 mg, at least 0.2 mg, at least 0.5 mg, at least 1 mg, at least 1 .5 mg, at least 2 mg, at least 3 mg, at least 4 mg or more than 4 mg per dose.

[0195] In some embodiments, the treatment involves intravitreal administration of between 0.05 mg to 10 mg, between 0.1 mg to 10 mg, between 0.1 mg to 5 mg, between 0.12 mg to 5 mg, between 0.2 mg to 5 mg per dose, between 0.2 mg to 4 mg per dose, between 0.5 mg to 4 mg or between 1 mg and 4 mg per dose. In some embodiments, the treatment involves intravitreal administration of a high dose of between 1 mg to 10 mg Link_TSG6 polypeptide per dose. In some embodiments, the treatment involves intravitreal administration of about 0.1 mg, about 0.05 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg or about 10 mg Link_TSG6 polypeptide per dose.

[0196] In some embodiments, the treatment involves intravitreal administration of one dose (e.g. a dose as described hereinabove, e.g. a dose of 0.05 mg to 10 mg) of Link_TSG6 polypeptide once per week, once every two weeks, once every three weeks, once every 4 weeks or once every 8 weeks, preferably every 4 or 8 weeks. By way of example, in some embodiments, the treatment involves intravitreal administration of between 0.1 mg to 5 mg Link_TSG6 polypeptide once per week, once every two weeks, once every three weeks, once every 4 weeks or once every 8 weeks, preferably every 4 or 8 weeks. In some embodiments, the treatment involves intravitreal administration of at least 0.05 mg, at least 0.06 mg, at least 0.08 mg, at least 0.1 mg, at least 0.12 mg, at least 0.15 mg, at least 0.2 mg, at least 0.5 mg, at least 1 mg, at least 1 .5 mg, at least 2 mg, at least 3 mg or at least 4 mg of Link_TSG6 polypeptide, once per week, once every two weeks, once every three weeks, once every 4 weeks, or once every 8 weeks, preferably every 4 or 8 weeks.

[0197] Topical Administration

[0198] For topical administration of the Link_TSG6 polypeptide via an eye drop a dose can be considered as 1 eye drop. Each drop may be around 50 pl.

[0199] Appropriate human dosage values can be calculated from effective mouse dosages by 15x scaling based on corneal surface area. Accordingly, in some embodiments, the treatment involves topical administration of between 100 pg to 500 pg Link_TSG6 polypeptide per dose.

[0200] In some embodiments, the treatment involves topical administration of at least 100 pg, at least 150 pg, at least 200 pg, at least 250 pg, at least 300 pg, at least 350 pg, at least 400 pg, at least 450 pg, or more than 450 pg per dose.

[0201] In some embodiments, the treatment involves topical administration of between 100 pg to 500 pg, between 100 pg to 400 pg, between 150 pg to 400 pg, or between 150 pg to 300 pg Link_TSG6 per dose. In some embodiments, the treatment involves topical administration of about 100 pg, about 150 pg, 8849846 about 200 pg, about 250 pg, about 300 pg, about 350 pg, about 400 pg, about 450 or about 500 pg per dose Link_TSG6 polypeptide per dose.

[0202] In some embodiments, the treatment involves topical administration of a dose of Link_TSG6 polypeptide (e.g. an eye drop having a dose for topical administration described hereinabove, e.g. a dose of 100 pg to 500 pg) at a frequency of 1x, 2x, 3x or 4x per day, preferably 2x per day. In some embodiments, the treatment involves topical administration of a dose of Link_TSG6 polypeptide (e.g. an eye drop having a dose for topical administration described hereinabove, e.g. a dose of 100 pg to 500 pg) less frequently than once per day, such as once per week, once every two weeks, once every three weeks or once per month (one time per calendar month, or one administration on average every 30 days). By way of example, in some embodiments, the treatment involves topical administration of between 100 pg to 500 pg Link_TSG6 polypeptide 1x, 2x, 3x, or 4x per day. In some preferred embodiments, the treatment involves topical administration of between 100 pg to 500 pg Link_TSG6 polypeptide 2x per day. In some embodiments, the treatment involves topical administration of at least 100 pg, at least 150 pg, at least 200 pg, at least 250 pg, at least 300 pg, at least 350 pg, at least 400 pg, or more than 450 pg Link_TSG6 polypeptide 1x, 2x, 3x, or 4x per day, preferably 2x per day.

[0203] Formulations / Compositions

[0204] The present disclosure also provides compositions comprising Link_TSG6. The Link_TSG6 described herein may be formulated as pharmaceutical compositions or medicaments for clinical use. Such compositions may comprise a pharmaceutically acceptable carrier, diluent, excipient or adjuvant. In some embodiments, a pharmaceutical composition as described herein consists of a Link_TSG6 polypeptide and a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

[0205] The medicaments and compositions may be formulated in fluid or solid form. Fluid formulations may be formulated for administration by injection to a selected region of the human or animal body. For example, a fluid formulation may be formulated for administration by subcutaneous injection, topical administration to the eye (such as an eye drop), or intravitreal injection. Preferably, medicaments and pharmaceutical compositions according to aspects of the present invention are formulated for subcutaneous administration.

[0206] In some cases, the Link_TSG6 is formulated with one or more other agents known to be useful for the treatment of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels. In some methods disclosed herein, the Link_TSG6 polypeptide is administered in combination with an anti-VEGF therapy. The anti-VEGF therapy may be e.g. aflibercept, brolucizumab, ranibizumab, bevacizumab or faricimab-svoa. Accordingly, formulations of Link_TSG6 may additionally comprise an anti-VEGF therapy. 8849846

[0207] Pharmaceutical compositions may be prepared using a pharmaceutically acceptable “carrier” composed of materials that are considered safe and effective. "Pharmaceutically acceptable" refers to molecular entities and compositions that are "generally regarded as safe", e.g., that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, loss, or change, of taste (ageusia) and the like, when administered to a human. In some embodiments, this term refers to molecular entities and compositions approved by a regulatory agency of the US federal or a state government, as the GRAS list under section 204(s) and 409 of the Federal Food, Drug and Cosmetic Act, that is subject to premarket review and approval by the FDA or similar lists, the U.S. Pharmacopeia or another generally recognised pharmacopeia for use in animals, and more particularly in humans.

[0208] The term “carrier” refers to diluents, binders, lubricants and disintegrants. Those with skill in the art are familiar with such pharmaceutical carriers and methods of compounding pharmaceutical compositions using such carriers.

[0209] The pharmaceutical compositions provided herein may include one or more excipients, e.g., solvents, solubility enhancers, suspending agents, buffering agents, isotonicity agents, antioxidants or antimicrobial preservatives. When used, the excipients of the compositions will not adversely affect the stability, bioavailability, safety, and / or efficacy of the active ingredients, i.e. Link_TSG6 used in the composition. Thus, the skilled person will appreciate that compositions are provided wherein there is no incompatibility between any of the components of the dosage form. Excipients may be selected from the group consisting of buffering agents, solubilising agents, tonicity agents, chelating agents, antioxidants, antimicrobial agents, and preservatives.

[0210] The Link_TSG6 polypeptide may be formulated as an eye drop. Formulations suitable for ocular administration include eye drops wherein the active compound is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active compound.

[0211] Eye drop formulations as disclosed herein may further comprise one or more of a preservative, antioxidant, stabilizer, tonicity modifier, viscosity modifier or buffer. Preferably, the eye drop formulation is a sterile eye drop formulation.

[0212] In some cases, the eye drop formulation contains between 2000 and 10000 pg / ml Link_TSG6. Such formulations are useful for delivering between around 100 pg and around 500 pg of Link_TSG6 per drop. The eye drop formulation may comprise between 2000 pg / ml and 10000 pg / ml, between 2000 pg / ml and 8000 pg / ml, between 3000 pg / ml and 8000 pg / ml, or between 3000 pg / ml and 6000 pg / ml. The eye drop formulation may contain at least 2000 pg / ml, at least 3000 pg / ml, at least 3500 pg / ml, at least 4000 pg / ml, at least 4500 pg / ml, at least 5000 pg / ml, at least 5500 pg / ml, at least 6000 pg / ml, at least 6500 pg / ml, at least 7000 pg / ml, at least 7500 pg / ml, at least 8000 pg / ml or more than 8000 pg / ml Link_TSG6 polypeptide. 8849846

[0213] In some aspects, the eye drop formulation may further comprise artificial tears. Artificial tears are lubricant eye drops. Artificial tears may contain one or more agents selected from carboxymethyl cellulose, polyvinyl alcohol, hydroxypropyl methylcellulose, hydroxypropyl cellulose, hyaluronan, water, salts and polymers, such as polyethylene glycol or polypropylene glycol.

[0214] Accordingly, a further aspect disclosed herein is an eye drop formulation comprising Link_TSG6 polypeptide. The eye drop formulation may comprise between 1500 pg / ml and 3500 pg / ml, between 1500 pg / ml and 3000 pg / ml, between 2000 pg / ml and 3000 pg / ml, or between 2400 pg / ml and 3000 pg / ml. Preferably, the eye drop formulation comprises between about 2400 pg / ml and about 3000 pg / ml. In some cases, the eye drop formulation comprises at least 2200, at least 2300 pg / ml, at least 2400 pg / ml, at least 2500 pg / ml, at least 2600 pg / ml, at least 2700 pg / ml, at least 2800 pg / ml, at least 2900 pg / ml, at least 3000 pg / ml or more than 3000 pg / ml. The eye drop formulation may further comprise prednisolone or artificial tears. The eye drop formulation may comprise a pharmaceutically acceptable carrier.

[0215] Kits

[0216] In some aspects of the present disclosure a kit of parts is provided. In some embodiments, the kit may have at least one container having a predetermined quantity of a Link_TSG6 polypeptide or composition described herein.

[0217] In some embodiments, the kit may comprise materials for producing a Link_TSG6 polypeptide or composition described herein.

[0218] The kit may provide the Link_TSG6 polypeptide or composition together with instructions for administration to a patient in order to treat wet AMD, macular oedema secondary to retinal vein occlusion (branch RVO or central RVO), diabetic macular oedema (DME) or myopic choroidal neovascularisation (myopic CNV).

[0219] In some embodiments the kit may further comprise at least one container having a predetermined quantity of another therapeutic agent (e.g. as described herein). In such embodiments, the kit may also comprise a second medicament or pharmaceutical composition such that the two medicaments or pharmaceutical compositions may be administered simultaneously or separately such that they provide a combined treatment for the specific disease or condition.

[0220] In some embodiments, the second therapeutic agent is an anti-VEGF therapy. Accordingly, in some embodiments the kit may provide the Link_TSG6 polypeptide or composition, an anti-VEGF therapy and instructions for administration to a patient in order to treat wet AMD, macular oedema secondary to retinal vein occlusion (branch RVO or central RVO), diabetic macular oedema (DME) or myopic choroidal neovascularisation (myopic CNV). 8849846

[0221] ***

[0222] The features disclosed in the foregoing description, or in the following claims, or in the accompanying drawings, expressed in their specific forms or in terms of a means for performing the disclosed function, or a method or process for obtaining the disclosed results, as appropriate, may, separately, or in any combination of such features, be utilised for realising the invention in diverse forms thereof.

[0223] While the invention has been described in conjunction with the exemplary embodiments described above, many equivalent modifications and variations will be apparent to those skilled in the art when given this disclosure. Accordingly, the exemplary embodiments of the invention set forth above are considered to be illustrative and not limiting. Various changes to the described embodiments may be made without departing from the spirit and scope of the invention.

[0224] For the avoidance of any doubt, any theoretical explanations provided herein are provided for the purposes of improving the understanding of a reader. The inventors do not wish to be bound by any of these theoretical explanations.

[0225] Any section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0226] Throughout this specification, including the claims which follow, unless the context requires otherwise, the word “comprise” and “include”, and variations such as “comprises”, “comprising”, and “including” will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0227] It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by the use of the antecedent “about,” it will be understood that the particular value forms another embodiment. The term “about” in relation to a numerical value is optional and means for example ± 10%.

[0228] Brief Description of the Figures

[0229] Embodiments and experiments illustrating the principles of the invention will now be discussed with reference to the accompanying figures.

[0230] Figure 1. Body weight of mice at 0-, 4-, 7- and 14-days post treatment with Link_TSG6 Low 0.12 pg (IVT), Link_TSG6 High 1.2 pg (IVT), PBS (IVT), Eylea (Aflibercept) (IVT), rhTSG-6 0.4 pg (IVT), PBS 8849846

[0231] (topical), Link_TSG6 low 1 pg (topical), Link_TSG6 high 10 pg (topical), PBS (SC), Link_TSG6 Low 13.75 pg (SC) and Link_TSG6 High 55 pg (SC).

[0232] Figure 2. Example image of FA scans taken immediately after CNV induction from retinal (upper row) and choroidal (lower) focus levels. Lasered sites that became leaky are outlined in dark grey, grey and white from choroidal level images (animal no. MU4519-007, treatment group Link_TSG6 IVT low 0.12 pg).

[0233] Figure 3. Example image of SD-OCT scans showing lasered sites outlined in dark grey, white and grey (animal no. MU4519-007, treatment group Link_TSG6 IVT low 0.12 pg).

[0234] Figure 4. Qualitative assessment of choroidal leak post CNV induction and (A) intravitreal treatment, (B) topical treatment or (C) subcutaneous treatment. Fisher’s exact test: **follow-up day 4, PBS vs aflibercept group, p=0.0014; *follow-up day 7, PBS vs aflibercept group, p=0.0167, **follow-up day 7, PBS vs Link TSG6 high group, p=0.0075.

[0235] Figure 5. CNV leak area at 4, 7 or 14 days post CNV induction and (A) intravitreal treatment, (B) topical treatment or (C) subcutaneous treatment. Kruskal-Wallis test, *PBS vs aflibercept, p=0.010; #PBS vs Link TSG6 high, p=0.028.

[0236] Figure 6. Representative images of isolectin Sustained choroidal flat-mounts from intravitreally treated groups. Scale bar = 100 pm.

[0237] Figure 7. Representative images of isolectin B4-stained choroidal flat-mounts from topical and subcutaneously treated groups. Scale bar = 100 pm.

[0238] Figure 8. Isolectin B4-immunoreactive choroidal area after (A) intravitreal treatment, (B) topical treatment and (C) subcutaneous treatment with PBS, Link_TSG6 low or Link_TSG6 high. Results are presented as mean ± SD.

[0239] Figure 9. Graphical presentation of the study design, intravitreal (IVT) treatment groups. FA: Fluorescein angiography; SD-OCT: spectral domain optical coherence tomography.

[0240] Figure 10. Graphical presentation of the study design, subcutaneous (SC) treatment groups. FA: Fluorescein angiography; SD-OCT: spectral domain optical coherence tomography.

[0241] Figure 11. Animal body weights during the study after (A) intravitreal treatment and (B) subcutaneous treatment. No statistically significant differences between the groups were found either in the IVT treated groups (time P<0.001 , treatment P = 0.14, time xtreatment P = 0.11) or in the SC treated groups (time 8849846

[0242] P<0.001 , treatment P = 0.59, time x treatment P = 0.55). Data are presented as mean ± sem from 10-12 animals / group.

[0243] Figure 12. Representative images from FA and SD-OCT scans from the IVT treated groups on Day 0 immediately after CNV model induction. CNV lesions are outlined in circles at the 5-minute timepoint after fluorescein administration in FA scans (left panel) and the corresponding lesions are marked in the Day 0 SD-OCT scans (right panel).

[0244] Figure 13. Representative images from FA and SD-OCT scans from the SC treated groups on Day 0 immediately after CNV model induction. CNV lesions are outlined in circles at the 5-minute timepoint after fluorescein administration in FA scans (left panel) and the corresponding lesions are marked in the Day 0 SD-OCT scans (right panel).

[0245] Figure 14. Representative images of leaky and non-leaky lesions on Day 4. Circles in the top panel mark leaky lesions and circles in the bottom panel mark non-leaky lesions. For the leaky lesion, a haze-like signal of the leakage was also seen at the retinal level. Example images are from animal LK13423-064 for the vehicle group (leaky lesions) and LK13423-070 for the aflibercept group (non-leaky ones).

[0246] Figure 15. The percentage of leaky lesions during the study for the IVT groups (A-C) and for the SC groups (D) are presented as mean values. The lesions were graded as being leaky (1) or non-leaky (0) from the FA scans. The data were analyzed by two-way ANOVA (Study Day x Treatment P = 0.07 in A, P = 0.19 in B, P = 0.81 in C and P < 0.001 in D). All doses of Link_TSG6, administered SC, showed significant reduction of the CNV leakage on Day 4 when compared to the vehicle (*, P < 0.05 for 13.75 pg and 55 pg; ***, P < 0.001 for 110 pg and 220 pg). marks significance between Eylea® and vehicle when the number of leaky and non-leaky lesions are analyzed with Chi-square test. N =9-12 mice per group.

[0247] Figure 16. The percentage of leaky and non-leaky lesions per group on Day 4 for the IVT groups (A-C) and for the SC groups (D) are presented. The data were analysed by one-sided Chi-square test. Eylea® significantly reduced the number of leaky lesions when compared to the vehicle (A). Link_TSG6 4 pg and 0.12 pg doses showed significant reduction in the number of leaky lesions compared to the vehicle (B). No differences were seen between the rhTSG-6 groups and the vehicle (C). All the subcutaneously injected Link_TSG6 doses significantly reduced the number of leaky lesions when compared to the vehicle (D). *, P < 0.05; **, P < 0.01 ; ***, P < 0.001 . N =26-36 lesions / group (9-12 mice / group).

[0248] Figure 17. The percentage of leaky and non-leaky lesions per group on Day 7 for the IVT groups (A-C) and for the SC groups (D) are presented. The data were analysed by one-sided Chi-square test. Link_TSG6 0.12 pg and 1 .2 pg doses showed significant reduction in the number of leaky lesions compared to the vehicle and (B). No differences were seen between the rhTSG-6 groups and the vehicle (C). Subcutaneously injected Link_TSG6220 pg and 55 pg doses significantly reduced the number of leaky lesions when compared to the vehicle (D). *, P < 0.05, N =26-36 lesions / group (9-12 mice / group). 8849846

[0249] Figure 18. The areas of the FA leaks at different study timepoints are presented as mean values for the IVT groups in A-C and for the SC groups in D. The lesions were graded as being leaky (1) or non-leaky (0) from the FA scans. The leakage area of the leaky lesions were measured from the FA scans and for the non-leaky lesions the area is considered as 0. The data were analysed with two-way ANOVA or mixed-effects analysis (Study Day x Treatment P = 0.44 in A, P = 0.03 in B, P < 0.01 in C and P < 0.001 D). rhTSG-6 0.4 pg dose had significantly bigger leakage areas when compared to the vehicle group on Day 4 (C). Link_TSG6 110 pg and 220 pg doses had significantly smaller leakage areas at Day 4 when compared to the vehicle (D). N =26-36 lesions / group (9-12 mice / group).

[0250] Figure 19. Sequences relevant to the present disclosure.

[0251] Examples

[0252] EXAMPLE 1 : Proof of Concept study: Link TSG6 in a mouse CNV model

[0253] 1.1 Treatment Groups and study design

[0254] The objective of this study was to define the pharmacologic efficacy of test compounds administered intravitreally (IVT), topically and subcutaneously (SC) in a mouse model of choroidal neovascularisation (CNV).

[0255] Treatment groups are described in Table 1 below.

[0256] Table 1: Treatment groups. IVT: Intravitreally-treated, T: Topically-treated twice daily (BID) starting on day 0 prior to CNV induction until day 14, SC: Subcutaneously-treated, once weekly starting on day 0.

[0257] 1.2 Materials and Methods

[0258] 1.2. 1 Animals

[0259] The body weight of all animals was monitored at baseline prior to CNV induction and at every imaging timepoint, i.e. days 4, 7 and 14 prior to sacrifice. 8849846

[0260] For all the procedures the animals were anesthetized with an injection of a mixture containing ketamine (30 mg / kg) (Ketaminol Vet 50 mg / ml; Intervet, Germany) and medetomidine (0.4 mg / kg) (Cepetor Vet 1 mg / ml; Vetmedic, Finland). Anesthesia was reversed by a2-antagonist for medetomidine (2.5 mg / kg) (Revertor Vet 5 mg / ml; Vetmedic).

[0261] 1.2.2 Treatment Administration

[0262] Intravitreal administration of test compounds

[0263] Single intravitreal injections were performed immediately after the CNV induction on day 0. Two pl were injected unilaterally into the right eye. Chloramphenicol ointment was applied after the intravitreal injection (Oftan Chlora, Santen Oy).

[0264] Intravitreal administration of aflibercept (Eylea®)

[0265] The reference compound aflibercept (Eylea®, Bayer Pharma AG) was purchased as a ready-to-use solution for intravitreal injections at a concentration of 40 mg / ml (formulated in 10 mM sodium phosphate, 40 mM sodium chloride, 0.03% polysorbate 20, and 5% sucrose, pH 6.2).

[0266] Aflibercept was administered intravitreally into the right eye (2 pl at 40 mg / ml) immediately after the CNV induction on day 0. Chloramphenicol ointment was applied after the intravitreal injection (Oftan Chlora, Santen Oy).

[0267] Topical administration of test compounds

[0268] Topical treatments were administered twice per day starting at 7 am and starting at 7 pm. Three pl per eye were applied unilaterally on the right eye by pipetting the compound on top of the cornea. All treatments were started on day 0 prior to the CNV induction and continued for the whole follow-up period up to day 14 when animals were sacrificed.

[0269] Subcutaneous injections

[0270] 100 pl of treatment compounds were injected subcutaneously once weekly starting on day 0.

[0271] 1.2.3 Induction of Choroidal Neovascularisation (CNV)

[0272] The anesthetized animals received a drop of 0.5% tropicamid (Oftan Tropicamid, Santen Oy), to dilate the pupils. A drop of Viscotears (Bausch & Lomb) was applied on the eye and a coverslip used to applanate the cornea. Three laser lesions were executed unilaterally on the right eye around the optic nerve head using a 532 nm diode laser (spot size: 100 pm; power: 100 mW; time: 100 ms; Iridex Corp.). The success on perforating the Bruch’s membrane was verified by fluorescein angiography (FA) and spectral domain optical coherence tomography (SD-OCT) in vivo imaging.

[0273] After the CNV induction the choroidal lesions were monitored by FA and SD-OCT at days 4, 7 and 14 post-CNV. 8849846

[0274] 1.2.4 Fluorescein Angiography (FA)

[0275] Vascular leakage at the choroid level was examined using a Heidelberg Spectralis HRA2 system (Heidelberg Engineering, Germany). Briefly, a drop of 0.5% tropicamid (Oftan Tropicamid, Santen Oy) was administered on the cornea of the anesthetized mouse, in order to dilate the pupils, and the mouse was placed onto the mouse holder. After aligning the optic nerve head at the retina, with the use of the infrared reflectance camera, a solution of 2.5% sodium fluorescein (Sigma-Aldrich, Finland) was administered as a sc injection (30 pl / 10 g). Consecutive fluorescent images were taken every 60 sec from the retinal and choroidal focus level for a period of 5 min after fluorescein administration.

[0276] 1.2.5 Spectral-Domain Optical Coherence Tomography (SD-OCT)

[0277] After the FA imaging the mouse was transferred to the SD-OCT system (Envisu R2200 SD-OCT, Bioptigen Inc. / Leica Microsystems, USA), where a retinal scan was performed. The scanned area covers a 1 .4 x 1 .4 mm2of the retina centered around the optic nerve. Each scan is composed of 100 B Scans each one composed of 1000 A Scans.

[0278] 1.2.6 Qualitative and Quantitative Analysis of Vascular Leakage

[0279] The lasered spots were qualitatively graded from FA images for evidence of vascular leak. OCT scans were used for additional confirmation. Quantitative analysis from the vascular leakage was performed by manually delimiting the leakage area from choroidal FA images by using the FIJI / lmageJ software (Schindelin et al. 2012 Nat Methods 9(7): 676-82).

[0280] 1.2.7 Animal Sacrifice and Tissue Collection

[0281] Mice were killed by anesthesia overdose and were transcardially perfused with 0.9 % NaCI solution (Control Variable Speed Pump, Watson-Marlow Pumps, UK). The eyes were enucleated and post fixed at 4 °C overnight in 4 % paraformaldehyde in 0.1 M phosphate buffer solution, pH 7.4. The next morning the eyes were washed twice with PBS and choroidal flat-mounts were prepared from each treated eye and from one contralateral / untreated eye from each treatment group as negative control.

[0282] Eight mice from each treatment group were processed for histological analysis. The additional animals per group, if any, were sacrificed as described.

[0283] 1.2.8 Histological Staining and Analysis

[0284] Choroidal flat-mounts were stained with fluorescein labeled Griffonia Simplicifolia Lectin I isolectin B4 (FL- 1201 , Vector Laboratories, USA) and mounted with Fluoroshield™ mounting medium (Sigma-Aldrich) on microscopic slides. Thereafter, choroidal samples were imaged using a fluorescence microscope (Leica Microsystems, Germany). The isolectin-positive areas were manually outlined, and the stained area was measured using the image processing software FIJI / lmageJ (Schindelin et al. 2012). 8849846

[0285] 1.2.9 Data Analysis

[0286] Quantitative data was graphed, analysed and presented as mean ± standard deviation (SD), or otherwise stated. Outlier data points were identified using the ROUT’S test with a false discovery rate of 1 %. Data was analysed using the GraphPad Prism software (v8.0.1 GraphPad Software. USA). Differences were considered statistically significant at the P < 0.05 level.

[0287] 1.3 Results

[0288] 1.3.1 Body Weight

[0289] The body weight of all animals remained similar throughout the whole study. However, aflibercept-treated group had significantly lower body weight at the baseline, follow-up day 4 and follow-up day 7 as compared to the mice treated with Link TSG6 low 13.75 .g (mixed-effects model, P<0.019) (Figure 1).

[0290] 1.3.2 Qualitative Analysis of CNV Lesions

[0291] The damage of Bruch’s membrane was qualitatively graded as being leaky or non-leaky from FA scans (Figure 2) and verified from SD-OCT scans (Figure 3).

[0292] The presence of leaky CNVs at different in vivo imaging timepoints is presented in Figure 4 for each route of administration separately: A) intravitreal, B) topical, and C) subcutaneous.

[0293] On the follow-up day 7, eyes that received intravitreal injections of Link_TSG6 at a high dose had significantly lower area of FA leak as compared to PBS injected eyes (P=0.0075) (Figure 4A). Topical and SC Link_TSG6 treatment also showed lower CNV grading at days 7 and 14 compared to PBS (Figures 4B and 4C).

[0294] 1.3.3 Quantitative Assessment of CNV Lesions

[0295] The CNV leak area was estimated from FA scans taken at different in vivo imaging timepoints (Figure 5A- 5C). Similarly as in the CNV grading, aflibercept administered intravitreally had a significantly lower area of FA leak at the follow-up day 4 as compared to PBS injected eyes (Kruskal-Wallis test, P=0.010) (Figure 5A). On the follow-up day 7, eyes that received intravitreal injections of Link_TSG6 at a high dose had significantly lower area of FA leak as compared to PBS injected eyes (P=0.028) (Figure 5A).

[0296] Topical and SC Link_TSG6 treatment also showed less FA leakage area compared to PBS control at days 7 and 14 (Figures 5B and 5C).

[0297] 1.3.4 Histology

[0298] Choroidal flat-mounts were stained using isolectin B4 (in green) to verify the presence of neovascularisation. Representative images from different treatment groups are shown in Figures 6 and 7. 8849846

[0299] Eyes that received PBS intravitreal injections had the highest area of isolectin B4 staining among all treatment groups and different routes of administration. No statistically significant differences were found when groups from different treatment routes were compared (Figure 8).

[0300] 1.4 Conclusions

[0301] Link TSG6 administered intravitreally at a high dose significantly reduced CNV formation and vascular leak on Day 7 post-CNV induction and reduced CNV formation on day 14. Link TSG6 given either topically or systemically diminished CNV pathology on days 7 and 14.

[0302] EXAMPLE 2: IVT or SC Administration of Link TSG6 in a mouse CNV model

[0303] 2.1 Treatment Groups and study design

[0304] The objective of this study was to evaluate the efficacy of test articles administered intravitreally or subcutaneously in the mouse laser-induced choroidal neovascularisation (CNV) model.

[0305] Treatment groups are described in Table 2 below. The study design is represented graphically in Figures

[0306] 9 and 10.

[0307] Table 2: Treatment groups. IVT: intravitreal administration SC: sub-cutaneous administration

[0308] 2.2 Materials and Methods

[0309] 2.2.1 Sample preparation

[0310] Link_TSG6 and rhTSG-6 were stored in -70 °C until the day of use. All treatment solutions were prepared fresh before each administration time point into 2 ml protein low binding Microtubes (Sarstedt AG & Co. KG, cat# 72.695.600) and used within 4 hours. In all the cohorts, half of the treatment groups were administered in the morning, and half in the afternoon. Aflibercept (Eylea®, Bayer Pharma AG) was stored in +4 °C and used undiluted (Group 2). Aqua Sterilista (B. Braun, cat# 105536) sterile water was used in the test article resuspensions. The 2 x PBS was prepared from Oxoid™ Phosphate Buffered 8849846

[0311] Saline Tablets (Thermo Scientific, cat# BR0014G) and sterile filtered. The daily treatment solutions were prepared as follows:

[0312] Intravitreal Treatment Groups:

[0313] Eylea® was aliquoted from the original vial fresh each morning and used undiluted (Group 2). Group 1 (vehicle) treatment solution was prepared by pipetting 30 pl of 2x PBS into a fresh tube. The test article aliquots (Groups 3-9, 10-13) were taken from the freezer, spun down at the speed of 20,000g, and prepared on cold beads under fume hood, using endotoxin free solutions.

[0314] One aliquot labelled ‘Gp 3 rhTSG66.4 pg’ was resuspended in 32 pl water and mixed gently (Group 3, 0.2 pg / pl in 32 pl). One aliquot labelled ‘Gp 4 rhTSG6 19.5’ pg was resuspended in 30 pl water and mixed gently (Group 4, 0.65 pg / pl in 30 pl). One aliquot labelled ‘Gp 5&7 Link_TSG6 18 pg’ was resuspended in 15 pl water and mixed gently, and 1 .5 pl of the solution was pipetted to a fresh vial, diluted to 15 pl of water, and mixed gently. Equal volumes of 2 x PBS were added to both vials to get concentrations of 0.06 pg / pl in 30 pl (Group 5) and 0.6 pg / pl in 27 pl (Group 7). One aliquot labelled ‘Gp 6&8 Link_TSG6 64 pg’ was resuspended in 16 pl water and mixed gently, and 2 pl of the solution was pipetted to a fresh vial, diluted to 20 pl of water, and mixed gently. Equal volumes of 2 x PBS were added to both vials to get concentrations of 0.2 pg / pl in 40 pl (Group 6) and 2 pg / pl in 28 pl (Group 8).

[0315] Subcutaneous treatment groups:

[0316] Group 9 (vehicle) treatment solution was prepared by mixing 330 pl water with 330 pl 2x PBS to get 660 pl of 1x PBS. One aliquot of each tube for Groups 10-13 (labelled ‘Gp 10 Link_TSG6 90.75 pg’; ‘Gp 11 Link_TSG6 363 pg’; ‘Gp 12 Link_TSG6 726 pg’; and ‘Gp 13 Link_TSG6 1452 pg’, respectively) were resuspended in 330 pl of water and mixed gently. Equal volume of 2 x PBS was added to each tube to get 660 pl of solution.

[0317] All solutions were kept on cold beads until injected and used within 4 hours.

[0318] 2.2.2 Treatment Administration

[0319] Figure 9 shows the study design for the intravitreal (IVT) treatment groups (groups 1-8). For these groups, treatments were administered once using blinded codes as 2 pl intravitreal injections unilaterally (oculus dexter; OD) on Day 0, within 10 minutes after CNV induction.

[0320] Anesthetized animals were placed under a stereoscope (Leica Microsystems), and a drop of iodine was applied on the cornea and allowed to spread evenly (Minims Povidione Iodine 5%, Laboratoire Chauvin). A small incision in the sclera / choroid exposing the vitreous chamber was performed using a 30G needle near the limbus. A microsyringe with a 33G needle (Hamilton Bonaduz AG, Bonaduz) attached was used to inject the solutions into the intravitreal space. The solution was injected into the intravitreal space for 10 seconds and the needle was kept in place for an additional 30 seconds before being removed to avoid 8849846 reflux of the solution. Chloramphenicol ointment was applied topically afterthe injection (Oftan Chlora, Santen Oy).

[0321] Figure 10 shows the study design for the subcutaneous (SC) treatment groups (groups 9-13). For these groups, the treatments were administered using blinded codes as subcutaneous injections in the flank (100 pl / mouse / time point). On Day 0, the test articles were administered within 10 minutes afterthe CNV induction. On Day 7, 14, and 21 , the treatments were administered in the morning prior to in vivo imaging.

[0322] All solutions were kept on cold beads until injected and used within 4 hours. The remaining test article solutions were stored at -70 °C.

[0323] 2.2.3 Body Weight

[0324] The body weight of animals was monitored prior to each in vivo procedure on Days 0, 4, 7, 14, 21 , and 28.

[0325] 2.2.4 Medical Care

[0326] Daily animal welfare checks were conducted. Five animals developed wounds due to fighting with cage mates (LK13423-125, -136, -146, -151 , -152, and -160). The wounds were treated with vetericyn hydrogel (Innovacyn, Inc., cat# 10006509) and / or Vetramil (Bfactory Health Products B.V., cat# 1314) when needed until healed.

[0327] Two animals (LK13423-69, and -116) had anesthesia recovery difficulties and were found with slight inactivity and / or weight loss 1 - 4 days after anesthesia. These animals were given subcutaneous hydration (ringer lactate), extra warmth and wet feed (pellets mixed with tap water) and recovered. One animal (LK13423-146) had dental abnormalities, and its teeth were cut regularly, and body weight maintenance followed.

[0328] 2.2.5 Anesthesia and Reversal

[0329] For all the procedures the animals were anesthetized with a subcutaneous injection of a mixture containing ketamine (30 mg / kg; Ketaminol Vet, 50 mg / ml, Intervet International B.V) and medetomidine hydrochloride (0.4 mg / kg; Cepetor Vet, 1 mg / ml, CP-Pharma Handelsgesellschaft mbH). The anesthesia was reversed by the a2-antagonist for medetomidine atipamezole (2.5 mg / kg; Revertor Vet 5 mg / ml, CP- Pharma Handelsgesellschaft mbH). Anesthetic reagents were diluted in physiological saline (0.9% NaCI, B. Braun Melsungen AG) for dosing.

[0330] All procedures were performed on heated surfaces to prevent hypothermia, and animals were placed on heated cages for the anesthesia recovery period. All the animals received 200 pl of sodium lactate solution (Ringer-Lactat Animalcare, Animalcare Limited) subcutaneously under anaesthesia for hydration. 8849846

[0331] 2.2.6 Spectral-Domain Optical Coherence Tomography (SD-OCT)

[0332] SD-OCT was performed using the SD-OCT system Envisu R2200 (Bioptigen Inc. / Leica Microsystems, USA). The scanned area covers a 1 .4 x 1 .4 mm2 of the retina centered around the optic nerve. Each scan is composed of 100 B-Scans, each one composed of 1000 A Scans.

[0333] SD-OCT imaging was performed unilaterally at baseline prior to CNV induction, and on Days 0, 4, 7, 14, 21 , and 28 post-induction.

[0334] 2.2.7 Fluorescein Angiography (FA)

[0335] The vascular leakage at the choroid level was examined using a Heidelberg Spectralis HRA2 system (Heidelberg Engineering). Briefly, a drop of 0.5% tropicamide (Oftan Tropicamid. Santen Pharmaceutical Co., Ltd.) was applied on the cornea of the anesthetized mouse to dilate the pupils. After aligning the optic nerve head at the retina level with the use of an infrared reflectance camera, the animal was injected subcutaneously a solution of 2.5% sodium fluorescein (Sigma-Aldrich) (30 pl / 10 g). Consecutive fluorescent images were taken every 60 seconds from retina and choroid level for 5 min after the fluorescein administration.

[0336] FA was performed unilaterally (OD) after CNV induction on Days 0, 4, 7, 14, 21 , and 28 post-induction.

[0337] 2.2.8 Qualitative and Quantitative Analysis of CNV Lesions

[0338] The lasered spots were qualitatively graded from FA images as following: 1) grade 0 when no leakage was seen from images taken in a 1-min interval until 5 min after fluorescein injection, 2) grade 1 when leakage was detected. Hyperfluorescent areas without fluorescein leakage were graded as grade 0. OCT scans were used for additional confirmation of Bruch’s membrane damage. FA scans were analyzed by a proprietary algorithm, which uses a combination of convolutional neural network (CNN) designed for semantic segmentation and traditional computer vision algorithms. The neural network was trained to recognize and quantify CNV lesions using a transfer learning approach. The results from the model were reviewed by a scientist blinded to the treatments.

[0339] 2.2.9 Animal Sacrifice and Tissue Collection

[0340] At the end of the study period, after the Day 28 imaging, the mice were sacrificed by decapitation under anesthesia. The treated and induced eyes (OD) from all the groups were enucleated, orientation marked and post-fixed in 4% PFA for an hour. 8849846

[0341] 2.2.10 Excluded Animals and Lesions

[0342] A total of 14 mice were excluded during the in vivo study period. In addition, in four animals, the CNV lesions developed larger retinal degeneration, enlarged lesions or merging two lesions, and were excluded from the analysis from all time points.

[0343] In addition to the excluded animals, some lesions were excluded from the analysis.

[0344] During the FA analyses the lesions are reviewed and excluded from the downstream analysis if exclusion criteria is met. The lesion grading was evaluated from the FA scans by a researcher blinded to the treatment groups. A total of 5 lesions were excluded from 5 animals due to unsuccessful Bruch’s membrane perforation. The excluded lesions were from Groups 1 IVT vehicle (LK13423-60 shot 6), Group 3, rhTSG-6, 0.40pg (LK13423-62 shot 9), Group 5, Link_TSG6, 0.12pg (LK13423-48 shot 9), Group 7, Link_TSG6 1 ,20pg (LK13423-40 shot 12), and Group 11 , Link_TSG6, 55pg (LK13423-143 shot 9).

[0345] Some individual lesions from individual animals were excluded from the FA area analysis from days 21 and 28 due to exceptionally large areas. In addition, some individual lesions from individual time points were excluded from the analysis due to errors in Al algorithm, e.g., poorly recognized lesion area. These occurred in animals LK13423-98 and LK13423-114, vehicle SC; LK13423-69, rhTSG-6 1 ,30pg; LK13423- 47, Link_TSG6 4.0pg and LK13423-59 rhTSG-6, 0.40pg.

[0346] 2.3 Results

[0347] The CNV was induced unilaterally into the right eye (OD) by perforating Bruch’s membrane using a diode laser. The test articles were administered either intravitreally (IVT) into the lasered eye (OD) (Groups 1-8) or subcutaneously (SC; Groups 9-13) within 10 minutes after the induction.

[0348] In the IVT animals, the body weights increased steadily and similarly across the study groups (Figure 11 A). All groups followed normal distribution. No statistically significant differences between the groups were found (time P < 0.001 , treatment P = 0.14, time x treatment P = 0.11). Data were analysed by Two- way ANOVA followed by Tukey’s multiple comparisons test.

[0349] In the SC treated animals, the body weights increased steadily and similarly across the study groups (Figure 11 B). One outlier was identified on Day 4 (ROUT method, LK13423-116, Group 12, Weight 18.7g) due to drastic weight drop after previous anesthesia time point, and the value was excluded from the analysis. No statistically significant differences between the groups were found (time P < 0.001 , treatment P = 0.59, time x treatment P = 0.55). Data were analysed by Mixed effects model followed by Tukey’s multiple comparisons test. In addition, there were no signs of adverse reactions to administered treatment as monitored by general observation of the study animals. 8849846

[0350] 2.3.1 Qualitative Analysis of CNV Lesions

[0351] Confirmation of successful model induction and follow-up of the model development was performed using fluorescein angiography (FA) and spectral domain optical coherence tomography (SD-OCT) on Days 0, 4, 7, 14, 21 , and 28. The success of the CNV induction was confirmed on Day 0 using FA and SD-OCT (Figure 12 and Figure 13). The lasered areas were qualitatively graded as being leaky (1) or non-leaky (0) from FA scans. Representative images of leaky and non-leaky lesions on Day 4 are presented in Figure 14.

[0352] The percentage of leaky lesions per eye (animal) was plotted for IVT treated groups (Figure 15A-15C) and for SC treated groups (Figure 15D). The percentage of leaky and non-leaky lesions per group were analysed separately for each time point and the data were analysed by one-sided Chi-square test.

[0353] On Day 4, IVT Eylea® significantly reduced the number of leaky lesions when compared to the vehicle (P < 0.05) (Figure 15A and Figure 16A). IVT Link_TSG6 0.12 pg and 4 pg doses showed significant reduction in the number of leaky lesions compared to the vehicle (P < 0.05) (Figure 15B and Figure 16B). No differences were seen between the IVT rhTSG-6 groups and the vehicle (Figure 15C and Figure 16C). All SC Link_TSG6 doses significantly reduced the percentage of leaky lesions when compared to the vehicle (P < 0.05 for doses 13.75 pg and 55 pg and P < 0.001 for 110 pg and 220 pg) (Figure 16D).

[0354] On Day 7, IVT Link_TSG6 0.12 pg and 1 .2 pg doses showed significant reduction in the number of leaky lesions when compared to the vehicle (P < 0.05) (Figure 17B). No differences were seen between the IVT rhTSG-6 groups and the vehicle (Figure 17C). SC Link_TSG6 55 pg and 220 pg doses significantly reduced the number of leaky lesions when compared to the vehicle (P < 0.05) (Figure 17D).

[0355] 2.3.2 Quantitative Analysis of CNV Lesions

[0356] Leakage areas from each CNV lesion were measured from FA scans (Figure 18A-18D). For all non-leaky lesions the leakage area was set to 0 mm2in all group comparisons.

[0357] On Day 4, subcutaneously administered Link_TSG6 110 pg and 220 pg dose groups had a smaller leakage area when compared to the vehicle (P < 0.001) (Figure 18D).

[0358] 2.4 Conclusions

[0359] Surprisingly, all SC Link_TSG6 doses significantly reduced the number of leaky lesions on Day 4. Only SC Link_TSG6 doses of 55 pg and 220 pg had a significant reduction on Day 7 post-lasering as compared to the vehicle data. In addition, two of the highest doses (110 pg and 220 pg) of SC Link_TSG6 significantly reduced the leakage area on Day 4 post-lasering as compared to the vehicle group. This mouse CNV model resolves overtime (Figure 15A), so these results at the earlier timepoints (e.g. 4 and 7 days) are the most indicative of a therapeutic effect. 8849846

[0360] As expected, IVT Aflibercept (Eylea®) significantly reduced the number of leaky lesions at Day 4 post- lasering but lacked the efficacy at later timepoints of the study, Days 7-28, as compared to the vehicle group.

[0361] IVT Link_TSG6 at the doses of 0.12 pg and 4 pg showed significant reduction in the number of leaky lesions on Day 4 post-lasering, and IVT Link_TSG6 doses of 0.12 pg and 1 .2 pg showed significant reduction on Day 7 as compared to the vehicle data at those timepoints. No significant differences were found between IVT full-length TSG-6 (rhTSG-6) groups and the vehicle at any timepoint analyzed.

[0362] While we have seen effects of Link_TSG6 vs full-length TSG-6 in models of dry-eye disease and osteoarthritis (as described in the WO 2021 / 013452 A1 and WO2022157181), the mechanism of action in ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels is likely to be completely different from these other models.

[0363] In ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels Link_TSG6 is inhibiting choroidal neovascularisation (CNV), i.e., angiogenesis.

[0364] Full-length TSG6 was previously shown to reduce CNV, reduce VEGF and reduce CCL2 and CCR2+ cells (e.g., macrophages) (Kim et al., Sci Rep (2015) 5:11872). However, the effects described herein are surprising because full-length TSG-6 has an enzymatic function, leading to the covalent modification of hyaluronan, whereas Link_TSG6 completely lacks this activity (Getting et al. (2002) J. Biol. Chem. 277:51068-51076).

[0365] However, we unexpectedly found that Link_TSG6 is effective at treating ocular disorders characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, and working better than full- length TSG6. This could be because it’s smaller (11-kDa vs ~35-kDa) and more positively charged (pl of ~10 vs ~7) enabling easier movement from vitreal compartment to the choroidal blood vessels in the back of the eye. Link_TSG6 and full-length TSG6 also differ in their hyaluronan-binding properties (Link_TSG6 binds more weakly and without cooperative interactions) and have a differential effect on the crosslinking of this polysaccharide (Baranova et al., J Biol Chem. 2011 Jul 22; 286(29):25675-86).

[0366] EXAMPLE 3: Link TSG6 treatment of wet AMD in humans

[0367] The objective of this study is to evaluate the efficacy of Link_TSG6 administered subcutaneously in human patients suffering from ocular disorders characterised by macular oedema.

[0368] Patients were selected based on a previous diagnosis of wet AMD, retinal vein occlusion, diabetic macular oedema and myopic choroidal neovascularisation. Alternatively, patients were selected based on evidence of, or susceptibility to macular oedema, such as:

[0369] (i) distorted and reduced (i.e. wavy, fuzzy or blurry) vision, 8849846

[0370] (ii) difficulty recognizing familiar faces,

[0371] (iii) a dark, empty area or blind spot in the centre of vision,

[0372] (iv) complete loss of central vision,

[0373] (v) increased retinal thickness, and / or

[0374] (vi) a family history of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels (e.g. a family history of wet AMD).

[0375] Subjects received either Link_TSG6 or vehicle (PBS) control treatment. A subset of each treatment group received intravitreal administration and a subset received subcutaneous administration.

[0376] Treatments were administered at a frequency of 1x per month, 1x per 8 weeks or 1x per 12 weeks. Patients receiving Link_TSG6 were administered a dose of 0.05 mg to 200 mg of Link_TSG6 per dose.

[0377] Following treatment, patients were monitored for symptoms of macular oedema using eye examinations and OCT.

[0378] References

[0379] A number of publications are cited above in order to more fully describe and disclose the invention and the state of the art to which the invention pertains. Full citations for these references are provided below. The entirety of each of these references is incorporated herein.

[0380] Baranova NS, Nileback E, Haller FM, Briggs DC, Svedhem S, Day AJ, Richter RP. The inflammation- associated protein TSG-6 cross-links hyaluronan via hyaluronan-induced TSG-6 oligomers. J Biol Chem. 2011 Jul 22;286(29):25675-86. doi: 10.1074 / jbc.M111 .247395. Epub 2011 May 19. PMID: 21596748; PMCID: PMC3138277.

[0381] Chi-Chao Chan, Xiaoguang Cao, Defen Shen, Yujuan Wang, Jun Zhang, Joo Y. Oh, Darwin J.

[0382] Prockop, Jingsheng Tuo; Anti-inflammatory Recombinant TSG-6 Stabilizes The Progression Of Focal Retinal Degeneration In A Murine AMD Model. Invest. Ophthalmol. Vis. Sci. 2012;53(14):1228.

[0383] Early Treatment Diabetic Retinopathy Study design and baseline patient characteristics. ETDRS report number 7. Ophthalmology. 1991 May;98(5 Suppl):741-56. doi: 10.1016 / s0161 -6420(13)38009-9. PMID: 2062510.

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Claims

1. 8849846Claims:1 . A Link_TSG6 polypeptide for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels.

2. A method of treating an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, the method comprising administering Link_TSG6 to a patient in need thereof.

3. Use of a Link_TSG6 polypeptide in the manufacture of a medicament for the treatment of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels.

4. The Link_TSG6 polypeptide for use according to claim 1 , the method of claim 2 or the use of claim 3, wherein the ocular disorder is wet AMD, macular oedema secondary to retinal vein occlusion (branch RVO or central RVO), diabetic macular oedema (DME) or myopic choroidal neovascularisation (myopic CNV).

5. The Link_TSG6 polypeptide for use, the method, or the use of claim 4, wherein the ocular disorder is wet AMD.

6. The Link_TSG6 for use according to claim 1 , 4 or 5, the method of claim 2, 4 or 5 or the use of claim 3, 4 or 5, wherein the Link_TSG6 is formulated in a composition for subcutaneous administration.

7. The Link_TSG6 for use according to claim 1 or 4-6, the method of claim 2 or 4-6, or the use of claim 3 or 4-6, wherein the treatment comprises subcutaneous administration of Link_TSG6 polypeptide.

8. The Link_TSG6 for use, the method or the use of claim 6 or 7, wherein the Link_TSG6 is administered subcutaneously into the abdomen, thigh or upper arm.

9. The Link_TSG6 for use, the method or the use of any one of claims 6 to 8, wherein the Link_TSG6 is administered subcutaneously at a frequency of 1x per week.

10. The Link_TSG6 for use, the method or the use of any one of claims 6 to 8, wherein the Link_TSG6 is administered subcutaneously at a frequency of 1x per month.11 . The Link_TSG6 for use, the method or the use of any one of claims 6 to 8, wherein the Link_TSG6 is administered subcutaneously at a frequency of 1x per 8 weeks.884984612. The Link_TSG6 for use, the method or the use of any one of claims 6 to 8, wherein the Link_TSG6 is administered subcutaneously at a frequency of 1x per 12 weeks.

13. The Link_TSG6 for use, the method or the use of any one of claims 6 to 12, wherein the Link_TSG6 is administered subcutaneously at a dose of 0.5 mg to 200 mg Link_TSG6 per dose.

14. The Link_TSG6 for use according to claim 1 , 4 or 5, the method of claim 2, 4 or 5 or the use of claim 3, 4 or 5, wherein the treatment comprises intravitreal administration of LINK_TSG6 polypeptide.

15. The Link_TSG6 for use, the method, or the use of claim 14, wherein the Link_TSG6 is administered intravitreally 1x per month.

16. The Link_TSG6 for use, the method, or the use of claim 14 or claim 15, wherein the Link_TSG6 is administered intravitreally at a dose of about 0.05 mg to 10 mg Link_TSG6 per dose.

17. The Link_TSG6 for use according to claim 1 , 4 or 5, the method of claim 2, 4 or 5 or the use of claim 3, 4 or 5, wherein the treatment comprises topical administration of LINK_TSG6 polypeptide to the eye.

18. The Link_TSG6 for use, the method, or the use of claim 17, wherein the treatment comprises administering LINK_TSG6 polypeptide topically two times per day.

19. The Link_TSG6 for use, the method, or the use of claim 17 or claim 18, wherein the Link_TSG6 is administered at a dose of 100 pg to 500 pg Link_TSG6 per dose.

20. The Link_TSG6 polypeptide for use according to any one of claims 1 and 4 to 19, the method of any one of claims 2 and 4 to 19, or the use of any one of claims 3 to 19, wherein the Link_TSG6 polypeptide comprises, consists, or consists essentially of (i) the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9, or (ii) an amino acid sequence having at least 80% identity to the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9.21 . The Link_TSG6 for use, the method or the use of claim 20, wherein the Link_TSG6 polypeptide comprises, consists, or consists essentially of (i) the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9, or (ii) an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 9.

22. The Link_TSG6 polypeptide for use according to any one of claims 1 and 4 to 21 , the method of any one of claims 2 and 4 to 21 , or the use of any one of claims 3 to 21 , wherein the Link_TSG6 polypeptide is administered in combination with an anti-VEGF therapy.884984623. A Link_TSG6 polypeptide for use in the treatment or prevention of an ocular disorder characterised by leakage of fluid and / or blood from abnormal or damaged blood vessels, wherein treatment with the Link_TSG6 polypeptide results in a reduction in the percentage of leaky lesions.

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