Annexin A5 protein as a combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents for use in the treatment of macular edema or retinal vein occlusion.

Abstract

The present invention relates to novel methods, uses, and compositions of annexin A5 protein as co-therapy with one or more anti-VEGF agents in the prevention or treatment of, for example, macular edema, retinal vein occlusion, and / or one or more diseases, disorders or conditions associated with and / or caused by macular edema or retinal vein occlusion, and / or other conditions suitable for treatment with anti-VEGF agents. In preferred embodiments, annexin A5 is administered systemically, for example, intravenously.

Description

[Technical Field]

[0001] The present invention relates to novel methods, uses, and compositions for enhancing anti-VEGF therapy and / or mitigating complications associated with the administration of anti-VEGF agents. Specific areas of interest include the prevention or treatment of macular edema (hereinafter also referred to herein as macular edema or ME), the prevention or treatment of retinal vein occlusion, and conditions related to either. [Background technology]

[0002] Research on vascular endothelial-derived growth factor (VEGF) has revolutionized our understanding of vascularization and angiogenesis between disease onset and physiological homeostasis (Apte et al, 2019, Cell, 176(6):1248-1264). The central role of VEGF in the pathogenesis of blind eye diseases has also become clear.

[0003] For example, macular edema is a pathological accumulation of fluid in the center of the retina. It is a complication of many retinal diseases, including, among others, diabetic retinopathy (DR), retinal vascular occlusion (RVO), and uveitis (UV) (Haydinger et al, 2023, Frontiers in Medicine, 10:1128811). Macular edema causes decreased vision and, if chronic or refractory, can lead to severe permanent vision impairment and blindness. Combined with the underlying pathology, the edema can progress to irreversible tissue damage with retinal cell death and permanent vision impairment.

[0004] In most cases, macular edema develops due to dysregulation of the blood-retinal barrier (BRB), a set of structures that, in a healthy state, tightly regulate the passage of proteins, salts, metabolites, and other solutes between the blood and retinal tissue. BRB dysregulation allows for the infiltration of retinal tissue by proteins and other solutes that are normally retained in the blood (Haydinger et al, 2023 (see above)). Increased osmotic pressure in the tissue drives fluid accumulation.

[0005] Current treatments target blood-retinal barrier (BRB) dysfunction and include vascular endothelial growth factor (VEGF) blockers, corticosteroids, and nonsteroidal anti-inflammatory drugs (NSAIDs) (Haydinger et al, 2023 (see above); Ip & Hendrick, 2018, Asia-Pacific Journal of Ophthalmology, 7(1):40-45). These treatments target vasoactive and inflammatory mediators that cause disruption of the blood-retinal barrier. Other treatments include targeted laser photocoagulation (Haydinger et al, 2023 (see above), Ip & Hendrick, 2018 (see above)).

[0006] The first-line therapy for macular edema associated with diabetic retinopathy (DR) or retinal vascular occlusion (RVO), as well as the treatment of RVO itself, is anti-VEGF therapy, which may be enhanced with the use of intravitreal corticosteroids and / or retinal laser photocoagulation (Ehlers et al. 2022, Ophthalmology, 129:88-99; Ehlers et al, 2017, Ophthalmology, 124:1412-23, and Yeh et al, 2015, Ophthalmology, 122:769-78). The treatment of macular edema secondary to neovascular AMD also heavily relies on the use of anti-VEGF therapy (Flaxel et al, 2020, Ophthalmology, 127:P1-P65).

[0007] Saconni et al., 2019, F1000Research, Vol. 8, p. 1413, is titled "Emerging therapies in the management of macular edema." This paper mentions that the introduction of anti-vascular endothelial growth factor injections (ranibizumab and aflibercept) and dexamethasone implants into clinical practice has revolutionized the treatment of macular edema (ME) secondary to dr and recurrent vo-occlusive diseases (DRs) and rudimentary vo-occlusive diseases (RVOs). However, it notes that while fluocinolone acetonide implants are approved by the US FDA for the treatment of diabetic ME, they are not approved for RVOs. Therefore, the paper also mentions the need for new drugs in the treatment of resistant ME secondary to DRs and RVOs, and that brolucizumab and avisipalpegol have shown efficacy in preliminary studies and have a chance of being approved in the near future for the treatment of diabetic ME. Furthermore, it mentions that corticosteroids and nonbiological immunomodulators play an important role in ME secondary to uveitis.

[0008] The established standard of treatment for macular edema and / or RVO, and indeed several retinal diseases, is by direct delivery of the therapeutic agent to the eye via intravitreous injection (Haydinger et al, 2023 (see above), Ip & Hendrick, 2018 (see above), Cox et al., 2021, J. Clin. Med., 10(5):981; Hattenbach et al., 2023, Ophthalmol Sci., 3(3):100302). Since the retina is located at the back of the eye and surrounded by several protective layers, including the cornea, iris, and lens, such drugs are injected directly into the eye. These layers can make it difficult for drugs to reach the retina if they are administered by other routes. When a drug is injected into the eye, it is delivered directly to the retina and can act locally to target the cause of the problem. For example, anti-VEGF drugs are injected into the eye to block the action of VEGF-A, a protein that promotes the growth of abnormal blood vessels in the retina. By lowering the levels of VEGF-A, these drugs can help prevent the formation of new blood vessels and reduce inflammation and swelling of the eye. Injections into the eye are typically performed in an ophthalmologist's office or an outpatient surgical center. After the injection, the patient is typically monitored for a short period to ensure there are no adverse effects from the drug.

[0009] Targeting VEGF by intravitreal injection of monoclonal antibodies or high-affinity decoy receptors is an effective means for the treatment of most diseases associated with macular edema, including exudative age-related macular degeneration (Heier et al., 2012, Ophthalmology, 119:2537-48), diabetic macular edema (Nguyen et al., 2012, Ophthalmology, 119:789-801; Korobelnik et al, 2014, Ophthalmology, 121:2247-54), and macular edema caused by central retinal vein occlusion (CRVO) (Brown et al., 2010, Ophthalmology, 117:1124-33), branch retinal vein occlusion (BRVO) (Campochiaro et al., 2010, Ophthalmology, 117:1102-12), and uveitis (Mackensen et al., 2008, Retina, 28:41-5).

[0010] However, anti-VEGF therapy has concerns about ocular and systemic toxicity and requires long-term frequent reinjections (Nguyen et al., 2010, Ophthalmology, 117:2146-51, Campochiaro et al., 2010, Ophthalmology, 117:2387-94, Saint-Geniez et al, 2008, PLoS One., 3:e3554). Potential complications associated with intravitreal injection of anti-VEGF therapy include infectious endophthalmitis, sterile intraocular inflammation, and particularly retinal vasculitis caused by brolucizumab (Cox et al., 2021 (mentioned above)), and transient intraocular pressure (IOP) spikes (Levin et al., 2021, J Glaucoma, 30:1019-26).

[0011] Infectious endophthalmitis remains one of the most serious complications associated with anti-VEGF therapy. The reported incidence after injection varies from 0.008 to 0.092%, and despite its low incidence, a large number of repeated anti-VEGF injections mean that they account for a large proportion of cases of infectious endophthalmitis (Cox et al., 2021 (cited above)).

[0012] Sterile intraocular inflammation (SII), also known as pseudophthalmitis, is characterized by acute-onset intraocular inflammation without infection that resolves without antibiotic treatment. The reported incidence of SII associated with anti-VEGF varies from 0.02 to 0.37% depending on the study, and symptoms usually appear between 24 hours and 7 days after the stimulating injection. Symptoms can consist of blurred vision, floaters, pain, and photophobia. Pain is present in up to 46% of patients and its presence is significantly associated with severe vitreous or anterior chamber (AC) inflammation. Photophobia occurs in 4 - 19% of patients. In addition, visual acuity often drops significantly from its baseline at onset but often returns to pre-injection levels after the inflammation has resolved. Intraocular inflammation is present at the time of examination in virtually all cases. This can consist of vitritis, AC reaction, or most commonly both, and additional less common clinic findings include hypopyon, fibrin, corneal deposits, corneal edema, conjunctival congestion, and conjunctival edema (Cox et al., 2021 (cited above)).

[0013] Continuous anti-VEGF therapy by intravitreal injection is the current standard treatment for the treatment of macular edema (Ip & Hendrick, 2018 (cited above)). Hattenbach et al., 2023 (cited above) reported that although effective, some patients with RVO require frequent long-term injections and monitoring to maintain the recovery achieved during the initial intensive treatment. For example, a well-established approach is to inject anti-VEGF intravitreally monthly, repeated over several months or years (Apte et al., 2019 (cited above)).

[0014] However, Hattenbach et al., 2023 (mentioned above), in contrast to clinical trial results, suggest that data from clinical-setting studies indicate that many patients with RVO (both BRVO and CRVO) do not achieve the recovery observed in clinical trials, at least in part, because the frequency of anti-VEGF injections is below optimal. A recent analysis reported that patients with CRVO from clinical-setting studies were, on average, monitored less frequently, received fewer anti-VEGF injections, and did not achieve the visual recovery observed in patients with CRVO from clinical trials. Such reduced efficacy in clinical settings is likely to reflect the high burden (and resulting reduced adherence) associated with RVO treatment, including the need for regular anti-VEGF injections and appointments, as well as the associated costs.

[0015] Nevertheless, the indications for intravitreal injection are expanding, and it is recognized that an increasing number of patients are likely to be affected by the acute and chronic effects of intravitreal injection (Levin et al., 2021 (see above)). However, current treatments for macular edema or RVO have not demonstrated a reliable method for directly improving perfusion. Rather, current strategies focus on minimizing the impact of RVO outcomes and minimizing vision loss due to macular edema and neovascularization (Ip & Hendrick, 2018 (see above)).

[0016] Apte et al., 2019 (mentioned above), reported high interest and relevance in therapeutic approaches that complement VEGF-based strategies, and that the desired goal of multi-target approaches is to find a combination that can reduce the high therapeutic burden of frequent intraocular injections, improve visual outcomes, and prevent continuous vision loss from atrophic neurodegeneration. However, Apte et al., 2019 (mentioned above), reported that several strategies targeting novel pathways have so far failed to demonstrate efficacy. Similarly, Hattenbach et al., 2023 (mentioned above), reported that the need for more effective and sustainable therapeutic options for RVO to reduce the therapeutic burden without compromising visual outcomes remains unmet.

[0017] This is the background to the need for improved treatments for macular edema and related conditions, including RVO, in this technological field. In particular, there is a need for effective therapies that can be administered to the target patient without requiring direct injection into the eye (especially multiple repeated injections). [Overview of the project]

[0018] Surprisingly, against this background, Annexin A5 has been found to be effective in enhancing the form of anti-VEGF therapy and reducing the burden of repeated anti-VEGF administration, thereby providing an opportunity to reduce complications and / or side effects associated with repeated administration of anti-VEGF therapeutic agents. Furthermore, by reducing the need for repeated anti-VEGF administration, it is possible to reduce the opportunity for subjects to acquire decreased response and / or tolerance to the ongoing anti-VEGF treatment regimen.

[0019] Furthermore, the applicants found that when annexin A5 protein is administered systemically to subjects (e.g., intravenously), it can preferentially localize to the occlusion site in subjects with retinal vein occlusion (RVO). They also found that annexin A5 has biological activity that modulates factors contributing to blood-retinal barrier (BRB) dysfunction in subjects with macular edema and / or RVO, as well as factors contributing to other therapeutic benefits in subjects with macular edema and / or RVO.

[0020] Accordingly, a first aspect of the present invention provides a molecule comprising or consisting of the annexin A5 protein for use in the prevention or treatment of macular edema or related conditions in a subject, wherein this use is in combination with an anti-vascular endothelial growth factor (anti-VEGF) agent. Optionally, the molecule consists of monomeric annexin A5.

[0021] In an alternative embodiment of the first aspect of the present invention, an anti-VEGF agent is provided for use in the prevention or treatment of macular edema or related conditions in a subject, wherein this use is in combination with a molecule containing or comprising the annexin A5 protein. Optionally, this molecule comprises monomeric annexin A5.

[0022] In an alternative embodiment of the first aspect, the present invention provides a method for treating or preventing macular edema or a related condition in a subject, the method comprising administering a prophylactic or therapeutic dose of a molecule containing or comprising annexin A5 protein to the subject, the subject also being treated with a prophylactic or therapeutic dose of an anti-VEGF agent. Optionally, the molecule comprises monomeric annexin A5.

[0023] In an alternative embodiment of the first aspect, the present invention provides a method for treating or preventing macular edema or a related condition in a subject, the method comprising administering a prophylactic or therapeutically effective dose of an anti-VEGF agent to the subject, the subject also being treated with a prophylactic or therapeutically effective dose of a molecule comprising or consisting of annexin A5 protein. Optionally, this molecule consists of monomeric annexin A5.

[0024] In an alternative embodiment of the first aspect, the present invention provides the use of a molecule containing or comprising the annexin A5 protein in the manufacture of a drug for the prevention or treatment of macular edema or related conditions in a subject, wherein the use is in combination with an anti-VEGF agent. Optionally, the molecule comprises monomeric annexin A5.

[0025] In an alternative embodiment of the first aspect, the present invention provides the use of an anti-VEGF agent in the manufacture of a drug for the prevention or treatment of macular edema or related conditions in a subject, the use of which is in combination with a molecule containing or comprising the annexin A5 protein, or with an effective prophylactic or therapeutic dose of an annexin A5 molecule. Optionally, the molecule comprises monomeric annexin A5.

[0026] In some embodiments of the first aspect of the present invention, a molecule containing or comprising the annexin A5 protein, or a composition containing the molecule, is present in the same composition as the anti-VEGF agent.

[0027] In other embodiments of the first aspect of the present invention, a molecule containing or comprising the annexin A5 protein, or a composition containing the molecule, is formulated separately from the anti-VEGF agent. In this case, the anti-VEGF and annexin A5 components can be administered to the subject simultaneously or sequentially.

[0028] If a molecule containing or comprising the annexin A5 protein (or a composition containing that molecule) is formulated separately from the anti-VEGF agent, the anti-VEGF agent may be administered directly to the eye. For example, the anti-VEGF agent may be administered by intravitreal injection.

[0029] In the case of sequential administration of annexin A5 component and anti-VEGF component, in some cases, the molecule containing or consisting of the annexin A5 protein ("annexin A5 component") is administered to the subject as the first round of administration of the annexin A5 component before the initiation of anti-VEGF therapy. In other cases, the molecule containing or consisting of the annexin A5 protein may be administered to the subject as the first round of administration of the annexin A5 component after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. For example, in the case of a form of anti-VEGF therapy (in the absence of annexin A5 co-therapy) requiring multiple rounds of anti-VEGF administration for the prevention or treatment of macular edema or related conditions in the subject, the annexin A5 component may be administered to the subject as the first round of administration of the annexin A5 component before the initiation of anti-VEGF therapy, or after the initiation of anti-VEGF therapy, but before multiple subsequent rounds of administration of one or more of the anti-VEGF components.

[0030] In some embodiments of the first aspect of the present invention, the subject may also receive a second round of administration of annexin A5. In some examples, annexin A5 is administered to the subject as a first round of administration, followed by a second round of administration, before the initiation of anti-VEGF therapy. In other examples, annexin A5 is administered to the subject as a first round of administration, before the initiation of anti-VEGF therapy, and then as a second round of administration, after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other examples, annexin A5 is administered to the subject as a first round of administration, followed by a second round of administration, in which case both the first and second rounds of administration of annexin A5 occur after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In some options, there is no round of anti-VEGF therapy between the first and second rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between the first and second rounds of annexin A5 administration. In other options, there may be two or more rounds of anti-VEGF therapy between the first and second rounds of annexin A5 administration.

[0031] In some embodiments of the first aspect of the present invention, the subject may also receive a third round of administration of annexin A5. In some examples, annexin A5 is administered to the subject as a first round of administration, followed by a second round of administration, and then a third round of administration, all before the initiation of anti-VEGF therapy. In other examples, annexin A5 is administered to the subject as a first round of administration, followed by a second round of administration, both before the initiation of anti-VEGF therapy, and annexin A5 is administered to the subject as a third round of administration after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other cases, annexin A5 is administered to the subject as the first round of annexin A5 administration before the initiation of anti-VEGF therapy, then as the second round of annexin A5 administration, and subsequently as the third round of annexin A5 administration, in which case both the second and third rounds of administration occur after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. In other cases, annexin A5 is administered to the subject as the first round of annexin A5 administration, followed by the second round of annexin A5 administration, and subsequently as the third round of annexin A5 administration, in which case all three rounds of annexin A5 administration occur after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. In some options, there is no round of anti-VEGF therapy between the first and second rounds of annexin A5 administration and / or between the second and third rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between the first and second rounds of annexin A5 administration and / or between the second and third rounds of annexin A5 administration.In other options, there may be two or more rounds of anti-VEGF therapy between the first and second rounds of administration of annexin A5 component and / or between the second and third rounds of administration of annexin A5 component.

[0032] While the administration of one, two, or three rounds of annexin A5 component is described above, it is understood that one or more further rounds of annexin A5 component may be used beyond the third round. In some cases, all rounds of annexin A5 component administration are performed before the initiation of anti-VEGF therapy. In other cases, the first, second, third, or any subsequent round of annexin A5 component administration may be performed before the initiation of anti-VEGF therapy, and one or more subsequent rounds of annexin A5 component administration may be performed after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other cases, the rounds of annexin A5 component administration are performed after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In some options, there are no rounds of anti-VEGF therapy between the first and second rounds of annexin A5 administration, between the second and third rounds of annexin A5 administration, between the third round and subsequent rounds of annexin A5 administration, and / or between any of the multiple subsequent rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between any one or more of the first and second rounds of annexin A5 administration, the second and third rounds of annexin A5 administration, the third round and subsequent rounds of annexin A5 administration, and / or between any of the multiple subsequent rounds of annexin A5 administration. In other options, there may be two or more rounds of anti-VEGF therapy between any one or more of the following: the first and second rounds of administration of annexin A5 component, the second and third rounds of administration of annexin A5 component, the third round and subsequent rounds of administration of annexin A5 component, and / or multiple subsequent rounds of administration of annexin A5 component.

[0033] Subjects to whom co-therapy is applied according to a first aspect of the present invention are preferably observably responsive to the first round and / or any subsequent rounds of administration of the annexin A5 component portion of the co-therapy. That is, subjects preferably demonstrate an observable reduction, stabilization, or improvement of the macular edema condition and / or related conditions as a result of one or more rounds of administration of the annexin A5 component of the co-therapy. Such observable reduction, stabilization, or improvement of the macular edema condition may relate to one, two, three, or four of the following features, for example: i. Visual acuity as evaluated by the Best-Corrected Visual Acuity (BCVA) test, at the discretion of the user. ii. Macular retinal thickness, iii. Macular volume, and iv. Retinal Area of ​​Non-Perfusion (RANP). Typically, an observable reduction, stabilization, or improvement (or each) of an observable deterioration may and / or be observable before the subject receives subsequent administration of anti-VEGF agents.

[0034] As a result of administering the annexin A5 component in the first round and / or any subsequent rounds of administration, it becomes possible to reduce, or even completely avoid, the frequency and / or number of subsequent rounds of administration of the anti-VEGF component following the initial dose to the subject for a specified period, without substantially reducing the prophylactic or therapeutic efficacy of anti-VEGF therapy with respect to macular edema or related conditions. This can be compared, for example, with a suitable control such as a subject or cohort of subjects who have never been treated with a molecule containing or consisting of the annexin A5 protein over the same specified period, but have received multiple doses of the anti-VEGF component (e.g., consecutive doses approximately every 28 days (e.g., ±7, 6, 5, 4, 3, 2, or 1 day)). The specified period may be, for example, approximately, at most, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months, or approximately, at most, or at least 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “approximately” may refer to a period of less than ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or 1% of the stated period. The specified period may be until the end of the therapeutic or preventive need for the subjects. In some embodiments, a cohort may be two or more subjects, approximately, at least, or more than 3, 4, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, or 100 subjects.

[0035] The preventive or therapeutic efficacy of anti-VEGF therapy for macular edema or related conditions can be evaluated with respect to one, two, three, or four of the following characteristics: v. Visual acuity as evaluated by the Best Corrected Visual Acuity (BCVA) test, at the discretion of the user. vi. Macular retinal thickness, vii. Macular volume, and viii. Retinal non-perfused area (RANP).

[0036] In an embodiment of the first aspect of the present invention, in which a molecule containing or comprising the annexin A5 protein is administered to the subject before the initiation of anti-VEGF therapy, the reduction in the frequency of administration of the anti-VEGF component following the initial administration of the annexin A5 component can refer, for example, to the period between the initial administration of the annexin A5 component to the subject and the first administration of the anti-VEGF component to the subject, i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months or more, or a period exceeding 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years, provided that the subject receives at least one administration of the anti-VEGF component during treatment. In this context, the term “about” can refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0037] For example, according to a first aspect of the present invention, after administering the annexin A5 component to the subject in the first round (and / or any subsequent rounds of administration of the annexin A5 component), the subject may subsequently be administered an anti-VEGF component. i. Within a 12-month period from the start of treatment with Annexin A5 molecule or composition, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose, ii. Within a period of 10 months from the start of treatment with Annexin A5 molecules or compositions, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose. iii. Within an 8-month period from the start of treatment with Annexin A5 molecule or composition, 8, 7, 6, 5, 4, 3, 2 or less than 1 dose, and / or iv. Six, five, four, three, two, or less than one dose within a six-month period from the start of treatment with Annexin A5 molecules or compositions.

[0038] Additionally or alternatively, in subjects receiving co-therapy with Annexin A5 according to the first aspect of the present invention, the reduction in the frequency of subsequent rounds of administration of the anti-VEGF component following the initial administration of the anti-VEGF component may refer, for example, to the period between the initial administration of the anti-VEGF component to the subject and the second administration of the anti-VEGF component to the subject, i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months or more, or a period exceeding 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “about” may refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0039] Additionally or alternatively, in subjects receiving co-therapy with Annexin A5 according to the first aspect of the present invention, the reduction in the frequency of subsequent rounds of administration of the anti-VEGF component following the initial administration of the anti-VEGF component may refer, for example, to the period between the second round and / or any subsequent round of administration of the anti-VEGF component and the subsequent round of anti-VEGF administration, i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months or more, or a period exceeding 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “about” may refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0040] For example, according to a first aspect of the present invention, after the initial administration of an anti-VEGF component to a subject, an anti-VEGF agent can be subsequently administered to the subject receiving co-therapy with annexin A5. i. Within a 12-month period from the start of treatment with Annexin A5 molecule or composition, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose, ii. Within a period of 10 months from the start of treatment with Annexin A5 molecule or composition, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose. iii. Within an 8-month period from the start of treatment with Annexin A5 molecule or composition, 8, 7, 6, 5, 4, 3, 2 or less than 1 dose, and / or iv. Six, five, four, three, two, or less than one dose within a six-month period from the start of treatment with Annexin A5 molecules or compositions.

[0041] Preferably, according to a first aspect of the present invention, a molecule containing or comprising annexin A5 protein (such as a molecule comprising monomeric annexin A5) may be administered systemically to a subject, preferably intravenously, subcutaneously, or intramuscularly, or by choroidal injection (as discussed, for example, in Wu et al, 2023, Pharmaceuticals, 16(9):1241, the contents of which are incorporated herein by reference). When administered intravenously, a molecule containing or comprising annexin A5 protein, or a composition comprising this molecule, may be used in the form of a sterile aqueous solution that may contain, for example, other substances, such as a salt or glucose sufficient to make the solution isotonic with blood. The aqueous solution should be preferably buffered as needed (preferably to a pH of 3 to 9). For example, optionally, the aqueous solution is buffered to a pH within the range selected from approximately pH 4 to approximately pH 8, approximately pH 5 to approximately pH 8, approximately pH 6 to approximately pH 8, approximately pH 6.5 to approximately pH 7.9, approximately pH 6.8 to approximately pH 7.7, approximately pH 7.0 to approximately pH 7.5, approximately pH 7.1 to approximately pH 7.4, approximately pH 7.1 to approximately pH 7.4, and optionally, approximately pH 7.1, 7.2, 7.3, or 7.4, most preferably approximately pH 7.2. In this context, the term "approximately" when applied to the nearest integer pH value may refer to a value with a pH unit of ±0.5, 0.4, 0.3, 0.2, or 0.1 of the stated value, while the term "approximately" when applied to a pH value with one decimal place may refer to a value with a pH unit of ±0.05, 0.04, 0.03, 0.02, or 0.01 of the stated value.

[0042] Preferably, according to the first aspect of the present invention, with respect to the administration of the annexin A5 component to the subject in each round, the dose of a molecule containing or consisting of the annexin A5 protein is administered to the subject for two or more consecutive days, such as three, four, five, six, seven or more consecutive days, and / or for at least two, three, four, five, six, or seven days within a period of seven days (one week) out of eight days, such as at least one week, two weeks, three weeks, four weeks or more.

[0043] Optionally, according to a first aspect of the present invention, with respect to each round of administration of the annexin A5 component to a subject, the daily dose of a molecule containing or consisting of annexin A5 protein (such as a molecule consisting of monomeric annexin A5) administered to the subject is a) a total daily dose of about 5 mg, about 4 mg, about 3 mg, about 2 mg or less than about 2 mg, 0.1 to 1.5 mg, or about 1 mg, and / or b) within a range selected from the group consisting of about 50 to 100 μg, about 40 to 80 μg, about 30 to 60 μg, about 20 to 40 μg, 1 μg to 30 μg, or less than about 20 μg per kg of body weight of the subject. As used herein, the term “about” may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0044] In a second aspect of the present invention, a molecule comprising or consisting of annexin A5 protein is provided for use in the prevention or treatment of retinal vein occlusion (RVO) or related conditions in a subject, wherein the use is in combination with an anti-vascular endothelial growth factor (anti-VEGF) agent, preferably, a dose of the molecule comprising or consisting of annexin A5 protein is administered to the subject systemically or by parachoroidal injection, and / or the molecule comprising or consisting of annexin A5 protein is administered to the subject daily for two or more consecutive days. Optionally, the molecule comprises monomeric annexin A5.

[0045] In an alternative embodiment of a second aspect of the present invention, an anti-VEGF agent is provided for use in the prevention or treatment of retinal vein occlusion (RVO) or related conditions in a subject, wherein the use is in combination with a molecule containing or comprising annexin A5 protein, preferably a dose of the molecule containing or comprising annexin A5 protein administered to the subject systemically or by parachoroidal injection, and / or the molecule containing or comprising annexin A5 protein administered to the subject daily for two or more consecutive days. Optionally, the molecule comprises monomeric annexin A5.

[0046] In an alternative embodiment of the second aspect, the present invention provides a method for treating or preventing a RVO or related condition in a subject, the method comprising administering a prophylactic or therapeutic dose of a molecule containing or comprising the annexin A5 protein to the subject, preferably the dose being administered to the subject systemically or by choroidal injection and / or the dose being administered to the subject daily for two or more consecutive days, the subject also being treated with a prophylactic or therapeutic dose of an anti-VEGF agent. Optionally, the molecule comprises monomeric annexin A5.

[0047] In an alternative embodiment of the second aspect, the present invention provides a method for treating or preventing a RVO or related condition in a subject, the method comprising administering a prophylactic or therapeutically effective dose of an anti-VEGF agent to the subject, the subject also being treated with a molecule containing or comprising annexin A5 protein in a prophylactic or therapeutically effective dose. The dose of the molecule containing or comprising annexin A5 protein is preferably administered to the subject systemically or by choroidal injection and / or daily for two or more consecutive days. Optionally, the molecule consists of monomeric annexin A5.

[0048] In an alternative embodiment of the second aspect, the present invention provides the use of a molecule containing or comprising monomeric annexin A5 protein in the manufacture of a drug for the prevention or treatment of RVO or related conditions in a subject, the use being in combination with an anti-VEGF agent. The molecule containing or comprising monomeric annexin A5 protein is preferably administered to the subject systemically or by choroidal injection and / or daily for two or more consecutive days. Optionally, the molecule comprises monomeric annexin A5.

[0049] In an alternative embodiment of the second aspect, the present invention provides the use of an anti-VEGF agent in the manufacture of a drug for the prevention or treatment of RVO or related conditions in a subject, the use of which is in combination with a molecule containing or comprising monomeric annexin A5 protein, or with an effective prophylactic or therapeutic dose of annexin A5 molecule. The molecule containing or comprising monomeric annexin A5 protein is preferably administered to the subject systemically or by parachoroidal injection and / or daily for two or more consecutive days. Optionally, the molecule consists of monomeric annexin A5.

[0050] In some embodiments of a second aspect of the present invention, a molecule containing or comprising the annexin A5 protein, or a composition containing the molecule, is present in the same composition as the anti-VEGF agent.

[0051] In another embodiment of the second aspect of the present invention, a molecule containing or comprising the annexin A5 protein, or a composition containing the molecule, is formulated separately from the anti-VEGF agent. In this case, the anti-VEGF and annexin A5 components can be administered to the subject simultaneously or sequentially.

[0052] If a molecule containing or comprising the annexin A5 protein (or a composition containing that molecule) is formulated separately from the anti-VEGF agent, the anti-VEGF agent may be administered directly to the eye. For example, the anti-VEGF agent may be administered by intravitreal injection.

[0053] In the case of continuous administration, in some cases of annexin A5 and anti-VEGF components, the molecule containing or consisting of the annexin A5 protein ("annexin A5 component") is administered to the subject as the first round of administration of the annexin A5 component before the initiation of anti-VEGF therapy. In other cases, the molecule containing or consisting of the annexin A5 protein may be administered to the subject as the first round of administration of the annexin A5 component after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. For example, in the case of a form of anti-VEGF therapy (in the absence of annexin A5 co-therapy) requiring multiple rounds of anti-VEGF administration for the prevention or treatment of RVO or related conditions in the subject, the annexin A5 component may be administered to the subject as the first round of administration of the annexin A5 component before the initiation of anti-VEGF therapy, or after the initiation of anti-VEGF therapy, but before multiple subsequent rounds of administration of one or more of the anti-VEGF components.

[0054] In some first and second embodiments of the present invention, the subject may also receive a second round of administration of annexin A5 component. In some examples, annexin A5 component is administered to the subject as a first round of administration, followed by a second round of administration, before the initiation of anti-VEGF therapy. In other examples, annexin A5 component is administered to the subject as a first round of administration, before the initiation of anti-VEGF therapy, and then administered as a second round of administration, after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other examples, annexin A5 component is administered to the subject as a first round of administration, followed by a second round of administration, in which case both the first and second rounds of administration of annexin A5 component occur after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In some options, there is no round of anti-VEGF therapy between the first and second rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between the first and second rounds of annexin A5 administration. In other options, there may be two or more rounds of anti-VEGF therapy between the first and second rounds of annexin A5 administration.

[0055] In some embodiments of a second aspect of the present invention, the subject may also receive a third round of administration of annexin A5. In some examples, annexin A5 is administered to the subject as a first round of administration, followed by a second round of administration, and then a third round of administration, all before the initiation of anti-VEGF therapy. In other examples, annexin A5 is administered to the subject as a first round of administration, followed by a second round of administration, both before the initiation of anti-VEGF therapy, and annexin A5 is administered to the subject as a third round of administration after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other cases, annexin A5 is administered to the subject as the first round of annexin A5 administration before the initiation of anti-VEGF therapy, then as the second round of annexin A5 administration, and subsequently as the third round of annexin A5 administration, in which case both the second and third rounds of administration occur after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. In other cases, annexin A5 is administered to the subject as the first round of annexin A5 administration, followed by the second round of annexin A5 administration, and subsequently as the third round of annexin A5 administration, in which case all three rounds of annexin A5 administration occur after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. In some options, there is no round of anti-VEGF therapy between the first and second rounds of annexin A5 administration and / or between the second and third rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between the first and second rounds of annexin A5 administration and / or between the second and third rounds of annexin A5 administration.In other options, there may be two or more rounds of anti-VEGF therapy between the first and second rounds of administration of annexin A5 component and / or between the second and third rounds of administration of annexin A5 component.

[0056] While the administration of one, two, or three rounds of annexin A5 component is described above, it is understood that one or more further rounds of annexin A5 component may be used beyond the third round. In some cases, all rounds of annexin A5 component administration are performed before the initiation of anti-VEGF therapy. In other cases, the first, second, third, or any subsequent round of annexin A5 component administration may be performed before the initiation of anti-VEGF therapy, and one or more subsequent rounds of annexin A5 component administration may be performed after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other cases, the rounds of annexin A5 component administration are performed after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In some options, there are no rounds of anti-VEGF therapy between the first and second rounds of annexin A5 administration, between the second and third rounds of annexin A5 administration, between the third round and subsequent rounds of annexin A5 administration, and / or between any of the multiple subsequent rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between any one or more of the first and second rounds of annexin A5 administration, the second and third rounds of annexin A5 administration, the third round and subsequent rounds of annexin A5 administration, and / or between any of the multiple subsequent rounds of annexin A5 administration. In other options, there may be two or more rounds of anti-VEGF therapy between any one or more of the following: the first and second rounds of administration of annexin A5 component, the second and third rounds of administration of annexin A5 component, the third round and subsequent rounds of administration of annexin A5 component, and / or multiple subsequent rounds of administration of annexin A5 component.

[0057] Subjects to whom co-therapy is applied according to a second aspect of the present invention are preferably observably responsive to the administration of the first round of administration of the annexin A5 component portion of the co-therapy and / or any subsequent rounds of administration. That is, subjects preferably demonstrate an observable reduction, stabilization, or improvement of the RVO status and / or related conditions as a result of any one or more rounds of administration of the annexin A5 component of the co-therapy. Such observable reduction, stabilization, or improvement of deterioration may relate to, for example, one, two, three, or four of the following features: ix. Visual acuity as evaluated by the Best Corrected Visual Acuity (BCVA) test, at the discretion of the user. x. Macular retinal thickness, xi. Macular volume, and xii. Retinal non-perfused area (RANP). Typically, observable improvements (or each) may be observable and / or may be observed before the subject receives subsequent administration of anti-VEGF agents.

[0058] As a result of administering the annexin A5 component in the first round and / or any subsequent rounds, it becomes possible to reduce, or even completely avoid, the frequency and / or number of subsequent rounds of administration of the anti-VEGF component following the initial dose to the subject for a specified period, without substantially reducing the prophylactic or therapeutic efficacy of anti-VEGF therapy with respect to RVO or related conditions. This can be compared, for example, to a suitable control such as a subject or cohort of subjects who have never been treated with a molecule containing or consisting of the annexin A5 protein over the same specified period but have received multiple doses of the anti-VEGF component (e.g., consecutive doses approximately every 28 days (e.g., ±7, 6, 5, 4, 3, 2, or 1 day)). The specified period may be, for example, about, up to, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months, or about, up to, or at least 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “approximately” may refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period. The specified period may be until the end of the therapeutic or preventive need for the subjects. In some embodiments, a cohort may be two or more subjects, approximately, at least, or more than three, four, five, ten, fifteen, twenty, twenty-five, thirty, forty, fifty, sixty, seventy, eighty, ninety, or one hundred subjects.

[0059] The preventive or therapeutic efficacy of anti-VEGF therapy for RVO or related conditions can be evaluated with respect to one, two, three, or four of the following characteristics: xiii. Visual acuity as evaluated by the Best Corrected Visual Acuity (BCVA) test, at the discretion of the user. xiv. Macular retinal thickness, xv. Macular volume, and xvi. Retinal non-perfused area (RANP).

[0060] In an embodiment of a second aspect of the present invention, in which a molecule containing or comprising the annexin A5 protein is administered to the subject before the initiation of anti-VEGF therapy, the reduction in the frequency of administration of the anti-VEGF component following the initial administration of the annexin A5 component can refer, for example, to the period between the initial administration of the annexin A5 component to the subject and the first administration of the anti-VEGF component to the subject, i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months or more, or a period of 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years, provided that the subject receives at least one administration of the anti-VEGF component during treatment. In this context, the term "about" can refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0061] For example, according to a second aspect of the present invention, after the initial round of administration of annexin A5 component to the subject (and / or any subsequent rounds of administration of annexin A5 component), an anti-VEGF component can be subsequently administered to the subject. i. Within a 12-month period from the start of treatment with Annexin A5 molecule or composition, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose, ii. Within a period of 10 months from the start of treatment with Annexin A5 molecules or compositions, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose. iii. Within an 8-month period from the start of treatment with Annexin A5 molecule or composition, 8, 7, 6, 5, 4, 3, 2 or less than 1 dose, and / or iv. Six, five, four, three, two, or less than one dose within a six-month period from the start of treatment with Annexin A5 molecules or compositions.

[0062] Additionally or alternatively, in subjects receiving co-therapy with Annexin A5 according to a second aspect of the present invention, the reduction in the frequency of subsequent rounds of administration of the anti-VEGF component following the initial administration of the anti-VEGF component may refer, for example, to the period between the initial administration of the anti-VEGF component to the subject and the second administration of the anti-VEGF component to the subject, i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months or more, or a period exceeding 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “about” may refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0063] Additionally or alternatively, in subjects receiving co-therapy with Annexin A5 according to a second aspect of the present invention, the reduction in the frequency of subsequent rounds of administration of the anti-VEGF component following the initial administration of the anti-VEGF component may refer, for example, to the period between the second round and / or any subsequent round of administration of the anti-VEGF component and the subsequent round of anti-VEGF administration, i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months or more, or a period exceeding 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “about” may refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0064] For example, according to a second aspect of the present invention, after the initial administration of an anti-VEGF component to a subject, an anti-VEGF agent can be subsequently administered to the subject receiving co-therapy with annexin A5. i. Within a 12-month period from the start of treatment with Annexin A5 molecule or composition, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose, ii. Within a period of 10 months from the start of treatment with Annexin A5 molecules or compositions, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose. iii. Within an 8-month period from the start of treatment with Annexin A5 molecule or composition, 8, 7, 6, 5, 4, 3, 2 or less than 1 dose, and / or iv. Six, five, four, three, two, or less than one dose within a six-month period from the start of treatment with Annexin A5 molecules or compositions.

[0065] Preferably, according to a second aspect of the present invention, the mode of systemic administration of a molecule containing or comprising annexin A5 protein (such as a molecule comprising monomeric annexin A5 protein) may be selected from intravenous, subcutaneous, or intramuscular administration, or administered by choroidal injection (for example, as discussed in Wu et al, 2023, Pharmaceuticals, 16(9):1241 (the contents of which are incorporated herein by reference)). When administered intravenously, a molecule containing or comprising annexin A5 protein, or a composition comprising this molecule, may be used in the form of a sterile aqueous solution that may contain, for example, other substances, such as a salt or glucose sufficient to make the solution isotonic with blood. The aqueous solution should be preferably buffered as needed (preferably to a pH of 3 to 9). For example, optionally, the aqueous solution is buffered to a pH within the range selected from approximately pH 4 to approximately pH 8, approximately pH 5 to approximately pH 8, approximately pH 6 to approximately pH 8, approximately pH 6.5 to approximately pH 7.9, approximately pH 6.8 to approximately pH 7.7, approximately pH 7.0 to approximately pH 7.5, approximately pH 7.1 to approximately pH 7.4, approximately pH 7.1 to approximately pH 7.4, and optionally, approximately pH 7.1, 7.2, 7.3, or 7.4, most preferably approximately pH 7.2. In this context, the term "approximately" when applied to the nearest integer pH value may refer to a value with a pH unit of ±0.5, 0.4, 0.3, 0.2, or 0.1 of the stated value, while the term "approximately" when applied to a pH value with one decimal place may refer to a value with a pH unit of ±0.05, 0.04, 0.03, 0.02, or 0.01 of the stated value.

[0066] More preferably, the dose of a molecule containing or comprising annexin A5 protein is administered to the subject daily for two or more consecutive days, systemically (by intravenous, subcutaneous or intramuscular administration, etc.) or by choroidal injection.

[0067] In a second embodiment of the present invention, with respect to the administration of the annexin A5 component to a subject in each round, the dose of a molecule containing or consisting of the annexin A5 protein is administered to the subject daily for at least two, three, four, five, six, or seven days within each seven-day (one-week) or eight-day period, and / or for at least two, three, four, five, six, or seven days within each eight-day (one-week) period, such as three, four, five, five, six, or seven days within each eight-day (one-week) period.

[0068] Optionally, according to a second aspect of the present invention, with respect to the administration of annexin A5 component to a subject, the daily dose of a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) administered to the subject is within a range selected from the group consisting of a) about 5 mg, about 4 mg, about 3 mg, about 2 mg or less than about 2 mg, 0.1 to 1.5 mg, or about 1 mg total daily dose, and / or b) about 50 to 100 μg, about 40 to 80 μg, about 30 to 60 μg, about 20 to 40 μg, 1 μg to 30 μg, or less than about 20 μg per kg of body weight of the subject. As used herein, the term “about” may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0069] The inventors have also found that the treatment and prevention of macular edema, retinal vein occlusion and / or one or more conditions associated therewith according to first and / or second embodiments of the present invention can be achieved by molecules containing or comprising substantially lower doses of annexin A5 protein (such as monomeric annexin A5 protein).

[0070] For example, with respect to each round of administration of the annexin A5 component to a subject, the subject may be administered a molecule containing or consisting of the annexin A5 protein in doses of approximately 0.01 μg / kg to approximately 10 μg / kg or approximately 0.01 μg / kg to approximately 1 μg / kg, and / or the subject may be administered a molecule in doses of approximately 0.001 mg to approximately 1 mg or 1 μg to approximately 100 μg. Where used herein, the term “approximately” may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0071] Optionally, according to a second aspect of the present invention, a molecule comprising or consisting of annexin A5 protein may be used for use in combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents in the prevention or treatment of a condition in a subject having, suspected of having, or at risk of having retinal vein occlusion (RVO) (e.g., diagnosed with retinal vein occlusion (RVO)), the condition being selected from the group consisting of macular edema, retinal ischemia, iris neovascularization (optionally, iris neovascularization characterized by increased cytokines such as increased VEGF), retinal neovascularization (optionally, retinal neovascularization characterized by increased cytokines such as increased VEGF), neovascular glaucoma, vitreous hemorrhage, rubeosis, and retinal detachment.

[0072] Optionally, according to the first and / or second aspects of the present invention, the retinal vein occlusion may be selected from, for example, central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), or hemi-retinal vein occlusion (HRVO).

[0073] Optionally, according to the first and / or second aspects of the present invention, the retinal vein occlusion may optionally be acute retinal vein occlusion, such as acute central retinal vein occlusion (CRVO), acute branch retinal vein occlusion (BRVO), or acute hemiretinal vein occlusion (HRVO).

[0074] Optionally, according to one embodiment of the first and / or second aspects of the present invention, the use or method may also be for the prevention or treatment of one or more diseases, disorders or conditions related to and / or caused (directly or indirectly) by macular edema and / or retinal vein occlusion.

[0075] Optionally, according to the first and / or second aspects of the present invention, the subject may be characterized by having about 1.0% to about 2% of red blood cells that are annexin A5 positive. In this context, the term “about” when applied to a value stated as the nearest integer may refer to a value that is ±0.5, 0.4, 0.3, 0.2, or 0.1 of the stated value, while the term “about” when applied to a value stated to one decimal place may refer to a value that is ±0.05, 0.04, 0.03, 0.02, or 0.01 of the stated value.

[0076] Optionally, according to first and / or second aspects of the present invention, the subject may be characterized by having an average number of monomeric annexin A5 binding sites of about 400 to about 1000 per red blood cell. In this context, the term “about” may refer to values ​​less than ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated value.

[0077] Optionally, according to first and / or second aspects of the present invention, the subject is typically a human subject, e.g., a human male and / or a human female. The subject may be an adult, or less typically, a young person, a child, or a newborn.

[0078] Optionally, according to the first and / or second aspects of the present invention, the subject is a human adult and / or having a body weight of about 50 kg to about 150 kg. Where used herein, the term “about” may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0079] Optionally, according to the first and / or second aspects of the present invention, the subject may be additionally or alternatively characterized by one or more of the following: a. The target group is people aged approximately 50 and over. b. The subjects are those who do not have or are not suspected of having cardiovascular disease (myocardial infarction, stroke, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis), or diseases related to cardiovascular disease (sickle cell anemia, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (e.g., Behçet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably diabetes mellitus and diabetes mellitus vera). c. The subjects are those who do not have a history of cardiovascular disease (myocardial infarction, stroke, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis) or cardiovascular disease-related diseases (sickle cell anemia, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (e.g., Behçet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably excluding diabetes mellitus and diabetes mellitus vera), d. The subjects are those who do not have a family history of cardiovascular disease (myocardial infarction, stroke, atherosclerosis, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis) or a disease related to cardiovascular disease (sickle cell anemia, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (e.g., Behçet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably excluding diabetes mellitus and diabetes mellitus vera), and / or e. The subject does not have, or is not suspected of having, one or more conditions selected from solid tumors, lymphomas and / or hematological malignancies, such as breast cancer, endometrial cancer, cervical cancer, ovarian cancer, lung cancer, head and / or cervical cancer, brain cancer, thyroid cancer, esophageal cancer, stomach cancer, colorectal and / or rectal cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, bone cancer, lymphoma (such as Hodgkin lymphoma and / or non-Hodgkin lymphoma), and / or hematological malignancies (such as acute lymphoblastic leukemia, acute myeloid leukemia, and / or chronic myeloid leukemia).

[0080] In one embodiment of the first and / or second aspects of the present invention, the dose of a molecule containing or comprising the annexin A5 protein is administered to the subject at least once per day. The dose of the annexin A5 molecule may optionally be administered to the subject at least once per day for at least two, three, four, five, six, or seven days within a period of seven days (one week) out of eight days, such as a period of one, two, three, five, six, or six or more consecutive days, and / or a period of at least one week, two, three, or four weeks or more.

[0081] In one embodiment of the first and / or second aspects of the present invention, the first round and / or any subsequent rounds of administration of a molecule containing or comprising annexin A5 protein may be administered systemically to the target vascular system, for example, by intravenous administration, or by choroidal injection. When administered intravenously, the molecule containing or comprising annexin A5 protein, or a composition comprising this molecule, may be used in the form of a sterile aqueous solution that may contain, for example, other substances, such as salts or glucose sufficient to make the solution isotonic with blood. The aqueous solution should be preferably buffered as needed (preferably to a pH of 3 to 9). For example, optionally, the aqueous solution is buffered to a pH within the range selected from approximately pH 4 to approximately pH 8, approximately pH 5 to approximately pH 8, approximately pH 6 to approximately pH 8, approximately pH 6.5 to approximately pH 7.9, approximately pH 6.8 to approximately pH 7.7, approximately pH 7.0 to approximately pH 7.5, approximately pH 7.1 to approximately pH 7.4, approximately pH 7.1 to approximately pH 7.4, and optionally, approximately pH 7.1, 7.2, 7.3, or 7.4, most preferably approximately pH 7.2. In this context, the term "approximately" when applied to the nearest integer pH value may refer to a value with a pH unit of ±0.5, 0.4, 0.3, 0.2, or 0.1 of the stated value, while the term "approximately" when applied to a pH value with one decimal place may refer to a value with a pH unit of ±0.05, 0.04, 0.03, 0.02, or 0.01 of the stated value.

[0082] Furthermore, with respect to the first and / or second aspects of the present invention (and similarly any other aspects of the present invention), molecules comprising or consisting thereof monomeric annexin A5 protein are disclosed as a therapeutic option herein, to be administered directly to the eye for the first round and / or (if applicable) any or all subsequent rounds of administration of the annexin A5 component of the therapy. For example, the annexin A5 molecule may be administered by choroidal injection or intravitreal injection. Such injections may be in the form of a pharmaceutical formulation or a medical device such as an implantable device. Such formulations or devices may be, for example, slow-release or continuous-release and / or reabsorbable formulations or devices. Slow or continuous drug release may have the ability to achieve therapeutic drug concentrations and therapeutic effects in the affected eye over a period of, for example, about 3 to about 180 days (e.g., about 4, 5 or 6 days, about 1, 2 or 3 weeks, about 1, 2, 3, 4, 5 or 6 months), or even longer. Alternatively, the annexin A5 molecule may be directly administered to the target retinal vein (such as the central retinal vein, retinal branch veins, and / or hemiretinal veins) by, for example, catheterization. However, this may be preferable to avoid.

[0083] The annexin A5 protein for use in the first and / or second embodiments of the present invention may comprise or consist of an annexin A5 protein, and optionally comprises or consists of a monomeric annexin A5 protein. Preferably, the above annexin A5 protein consists of a monomeric annexin A5 protein. Optionally, the above annexin A5 protein is selected from one or more of the following: a. Proteins that contain, are essentially derived from, or consist of the sequence of human annexin A5 protein (human monomeric annexin A5 protein and / or optionally, proteins containing or consisting of the sequence of SEQ ID NO: 1, with or without N-terminal methionine), b. Recombinant human monomer annexin A5 protein, etc. c. Mammalian orthologs of human annexin A5 protein, such as human monomer annexin A5 protein. Alleles or genetic variants of da), b), or c), Functional analogues or variants of human annexin A5 protein (such as monomeric annexin A5 protein) having the sequence defined by SEQ ID NO: 1, either containing or not containing eN-terminal methionine, are proteins that contain or consist of 50%, 60%, 70%, 75%, for example, more than 80%, 85%, 90%, or more more than 95%, or 99% of the amino acid sequence identity of human annexin A5 protein (such as monomeric annexin A5 protein), One of the biologically active fragments from fa) to e).

[0084] In some embodiments, the molecule containing annexin A5 protein for use in the first and / or second embodiments of the present invention may be a conjugate in which the annexin A5 protein (such as monomeric annexin A5 protein) is conjugated to a portion such as a non-proteinoid portion. Non-limitingly, this portion may be, for example, a therapeutic, prophylactic, diagnostic, prognostic, or theragnostic portion. In some embodiments, this portion may be a drug (such as a non-proteinoid drug) and / or a radioactive portion (for example, if the conjugate is suitable for use in radioimmunotherapy ("RIT").

[0085] Furthermore, compositions comprising molecules containing or consisting of the annexin A5 protein described herein, for use according to the first and / or second aspects of the present invention, are also provided herein. Such compositions may, for example, be unit dose compositions. A unit dose composition may contain, for example, a total dose of annexin A5 protein of about 5 mg, about 4 mg, about 3 mg, about 2 mg, or less than about 2 mg, such as about 0.1 to 1.5 mg, about 1 mg or less. In this context, the term “about” when applied to a unit dose value in mg described for the nearest integer may refer to a value that is ±0.5, 0.4, 0.3, 0.2, or 0.1 mg of the described value, while the term “about” when applied to a unit dose value in mg described to one decimal place may refer to a value that is ±0.05, 0.04, 0.03, 0.02, or 0.01 mg of the described value.

[0086] Optionally, the composition is a pharmaceutical composition, and further optionally, the composition further comprises one or more pharmaceutically acceptable carriers. In some embodiments, the composition comprises a molecule containing or comprising the annexin A5 protein and one or more pharmaceutically acceptable carriers. Optionally, in addition to use as a combination therapy with anti-VEGF agents, the composition is intended for use as a combination therapy with one or more other agents. Optionally, the annexin A5 molecule or its composition may be used, with or without one or more other agents, as a first-line treatment for the prevention or treatment of one or more diseases, disorders or conditions associated with and / or caused (directly or indirectly) by macular edema and / or retinal vein occlusion (RVO).

[0087] According to first and / or second aspects of the present invention, in some embodiments, molecules comprising or consisting of the annexin A5 protein or compositions comprising the annexin A5 protein are intended for and / or used as combination therapy with one or more other agents, and the subject is administered the annexin A5 molecule separately from the other agents in the same composition, simultaneously with or sequentially with a separate formulation containing the annexin A5 molecule.

[0088] Optionally, according to the first and / or second aspects of the present invention, a molecule containing or comprising the annexin A5 protein and an anti-VEGF agent are administered together, either simultaneously in separate doses or sequentially in separate doses. In some embodiments, when the administration is separate and sequential, the interval between the sequential administration of the annexin A5 molecule and the anti-VEGF agent is about 28 days or less, such as about 3 weeks or less, about 2 weeks or less, about 1 week or less, about 6, 5, 4, 3, 2 or 1 day or less (e.g., about 24, 28, 12, 6, 5, 4, 3, 2 or 1 hour or less). However, studies reported in this application have demonstrated that administration of annexin A5 can delay the need for anti-VEGF therapy for a much longer period, or even eliminate the need for anti-VEGF therapy entirely. Therefore, in other embodiments, when the administration is separate and sequential, the interval between the sequential administration of the annexin A5 molecule and the anti-VEGF agent is longer than about 28 days. For example, as described above, any given round of administration of annexin A5 may span multiple days and may include multiple repeated administrations to the subject. For example, with respect to any given round of administration of the annexin A5 molecule, the annexin A5 molecule may be administered to the subject for at least two consecutive days, such as three, four, five, six, seven or more consecutive days, and / or for at least two, three, four, five, six, or seven days within a period of seven days (one week) out of eight days, such as at least one week, two weeks, three weeks, four weeks or more. When annexin A5 administration spans multiple days, in some embodiments, the period between consecutive administrations of the annexin A5 molecule and the anti-VEGF agent is calculated relative to the start date of annexin A5 administration. In other embodiments, this period is calculated relative to the end date of annexin A5 administration.

[0089] In a third aspect, the present invention provides a composition (such as a unit dose composition) comprising a molecule comprising or consisting of the annexin A5 protein as defined above with respect to the first and / or second aspects of the present invention, Optionally, the composition may contain annexin A5 molecules (such as molecules consisting of monomeric annexin A5 protein) in amounts of approximately 50 μg / kg to approximately 100 μg / kg, approximately 40 μg / kg to approximately 80 μg / kg, approximately 30 μg / kg to approximately 60 μg / kg, approximately 20 μg / kg to approximately 40 μg / kg, approximately 1 μg / kg to approximately 30 μg / kg, or less than approximately 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to approximately 10 μg / kg or approximately 0.01 μg / kg to approximately 1 μg / kg, and / or The composition may optionally contain annexin A5 molecules (such as molecules consisting of monomeric annexin A5 protein) in a total amount of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg, and Optionally, the composition is intended for use in the prevention or treatment of one or more diseases, disorders or conditions in a subject that are associated with and / or caused (directly or indirectly) macular edema and / or retinal vein occlusion (RVO), and / or macular edema and / or retinal vein occlusion, as a combination therapy with an anti-vascular endothelial growth factor (anti-VEGF) agent. Where used herein, the term “about” may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0090] In an alternative embodiment of the third aspect, the present invention provides a method for treating or preventing one or more diseases, disorders or conditions in a subject (typically a human subject) that are related to and / or caused (directly or indirectly) by macular edema and / or retinal vein occlusion (RVO), the method comprising administering a prophylactic or therapeutic dose of a composition comprising or a molecule comprising annexin A5 protein (such as a molecule consisting of monomeric annexin A5 protein), wherein the subject is also treated with a prophylactic or therapeutic dose of an anti-VEGF agent. Optionally, the composition may contain annexin A5 molecules (such as molecules consisting of monomeric annexin A5 protein) in amounts of approximately 50 μg / kg to approximately 100 μg / kg, approximately 40 μg / kg to approximately 80 μg / kg, approximately 30 μg / kg to approximately 60 μg / kg, approximately 20 μg / kg to approximately 40 μg / kg, approximately 1 μg / kg to approximately 30 μg / kg, or less than approximately 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to approximately 10 μg / kg or approximately 0.01 μg / kg to approximately 1 μg / kg, and / or Optionally, the composition may contain annexin A5 molecules (such as molecules consisting of monomeric annexin A5 protein) in a total amount of about 5 mg, about 4 mg, about 3 mg, about 2 mg, or less than about 2 mg, for example, about 1 mg, about 0.1 to 1.5 mg, 0.001 mg to about 1 mg, or about 0.001 mg to about 0.1 mg. Where used herein, the term “about” may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0091] In one embodiment of a third aspect of the present invention, the composition is a unit dose composition, and optionally, treatment and / or prevention may include, or consist of, administering the complete unit dose, for example, in a single dose.

[0092] A composition used according to a third aspect of the present invention may be a pharmaceutical composition.

[0093] Optionally, a composition according to a third aspect of the present invention further comprises one or more pharmaceutically acceptable carriers.

[0094] For example, the composition may consist of an annexin A5 molecule (such as a molecule consisting of monomeric annexin A5 protein) and one or more pharmaceutically acceptable carriers.

[0095] In a third embodiment of the present invention, an effective prophylactic or therapeutic dose of an anti-VEGF agent is administered directly to the target eye. For example, the anti-VEGF agent may be administered by intravitreal injection.

[0096] In a fourth aspect, the present invention provides an intravitreal composition for sterile injection (such as a unit dose of an intravitreal composition for sterile injection) comprising or a molecule comprising annexin A5 protein (such as a molecule consisting of monomeric annexin A5), Typically, the composition is present in a sterile syringe, more preferably the syringe containing an amount of about 200 μL or less, preferably about 100 μL or less, and more preferably this amount is about 10 to about 90 μL, about 20 to about 80 μL, about 30 to about 70 μL, about 40 to about 60 μL, or about 50 μL (in this context, the term "about" refers to an amount that is ±4, 3, 2, or 1 μL of the stated value). Optionally, the composition may consist of molecules made up of monomeric annexin A5 protein in concentrations of (i) 50 μg / kg to approximately 100 μg / kg, approximately 40 μg / kg to approximately 80 μg / kg, approximately 30 μg / kg to approximately 60 μg / kg, approximately 20 μg / kg to approximately 40 μg / kg, approximately 1 μg / kg to approximately 30 μg / kg, or less than approximately 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to approximately 10 μg / kg, or approximately 0.01 μg / kg to approximately 1 μg / kg. (ii) a total amount of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg, and the term “approximately” as used in this context may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned. Optionally, the composition is intended for use in the prevention or treatment of one or more diseases, disorders or conditions in the subject that are associated with and / or caused (directly or indirectly) macular edema and / or retinal vein occlusion (RVO), and / or macular edema and / or retinal vein occlusion, and such use is intended as combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents.

[0097] A fourth aspect of the present invention may be presented in the form of a kit comprising the composition and one or more additional components.

[0098] For example, a kit may include a composition present in a sterile syringe and one or more sterile needles attached to it or included separately within the kit. Preferably, the needle or each needle may be a gauge selected from gauges 27, 28, 29, or more preferably 30 or 31. The needle or each needle may preferably have a length of about 0.5 to 0.62 inches (12.7 to 15.75 mm). Longer needles may increase the risk of retinal damage if the patient accidentally moves forward during the intravitreal injection procedure.

[0099] In another example, the kit may comprise one or more further compositions, each containing one or more antibiotics, for intravitreal injection and / or subsequent administration to a patient.

[0100] In a fourth embodiment of the present invention, the anti-VEGF agent is administered directly to the target eye. For example, the anti-VEGF agent may be administered by intravitreal injection.

[0101] In a fifth aspect of the present invention, a) Prevention or treatment of VEGF-related diseases, disorders or conditions in the subject, and / or prevention or treatment of diseases, disorders or conditions in the subject, accompanied by one or more administrations of anti-VEGF agents to the subject. b) Enhancement of anti-VEGF therapy, and / or, c) Reduction of complications associated with the administration of anti-VEGF agents. Molecules containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein or a molecule consisting of its conjugate) are provided for use in the above.

[0102] In some embodiments of a fifth aspect of the present invention, the disease, disorder, or condition is selected from macular edema, RVO, or a condition associated with macular edema or RVO. These and other diseases, disorders and / or conditions associated with VEGF are also described in Apte et al., 2019 (above) (the contents of which are incorporated herein by reference), and may include one or more of the following forms of cancer: tumor growth, metastasis and / or other forms (including focal, progressive, metastatic or recurrent colorectal cancer (CRC), metastatic NSCLC, recurrent glioblastoma, cervical cancer, certain recurrent epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer, metastatic renal cell carcinoma (RCC), gastric or gastroesophageal adenocarcinoma, hepatocellular carcinoma, thyroid cancer, pancreatic neuroendocrine tumors, gastrointestinal stromal tumors, soft tissue sarcoma, and medullary thyroid carcinoma), as well as noncancerous conditions such as retinopathy associated with certain blindness diseases, including age-related macular degeneration (AMD), diabetic retinopathy and hypertensive retinopathy.

[0103] In some embodiments, the augmentation of anti-VEGF therapy provided by a fifth aspect of the present invention may result in, for example, a higher or more complete preventive or therapeutic effect, or the achievement of a preventive or therapeutic endpoint, than could be achieved by anti-VEGF therapy alone. This may be, for example, a comparison with a subject receiving the same dose, frequency, and / or duration of anti-VEGF administration but not administration of annexin A5 molecules.

[0104] Additionally or alternatively, the augmentation properties of anti-VEGF therapy provided by the fifth aspect of the present invention may include, for example, reducing the frequency of repeated anti-VEGF administrations during the course of anti-VEGF therapy, reducing the required dose of anti-VEGF therapy, and / or shortening the duration for which anti-VEGF therapy is required, in order to obtain the same preventive or therapeutic endpoint as that achieved by anti-VEGF therapy alone. This may be, for example, a comparison between a subject treated according to the fifth aspect of the present invention and a subject treated with anti-VEGF therapy alone, or a subject receiving anti-VEGF monotherapy who achieved the same preventive or therapeutic endpoint.

[0105] In a fifth embodiment of the present invention, the anti-VEGF agent is administered directly to the target eye. For example, the anti-VEGF agent may be administered by intravitreal injection.

[0106] In cases of macular edema, RVO, macular edema or RVO-related conditions, and other ocular conditions, the preventive or therapeutic effect, or the preventive or therapeutic endpoint, may be determined by evaluating one, two, three, or four of the following characteristics: i. Optional improvement in visual acuity as assessed by the Best Corrected Visual Acuity (BCVA) test. ii. Increased macular retinal thickness, iii. Increase in macular volume, and iv. Increase in retinal non-perfusion areas (RANPs).

[0107] In some embodiments, the reduction of complications associated with the administration of anti-VEGF agents may include reduction or avoidance of ongoing anti-VEGF therapy, decreased response to anti-VEGF therapy, and / or tolerance to anti-VEGF therapy in subjects. For example, such an effect may be a comparison between subjects treated according to a fifth aspect of the present invention and subjects treated with anti-VEGF therapy alone, or between subjects receiving anti-VEGF monotherapy who reached the same preventive or therapeutic endpoint.

[0108] In some embodiments, the reduction of complications associated with the administration of anti-VEGF agents may include the reduction or avoidance of ocular and / or systemic toxicity in anti-VEGF therapy. This may be, for example, a comparison between subjects treated according to a fifth aspect of the present invention and subjects treated with anti-VEGF therapy alone, or a comparison between subjects receiving anti-VEGF monotherapy who achieved the same preventive or therapeutic endpoint.

[0109] In some embodiments, the reduction of complications associated with the administration of anti-VEGF agents may include the reduction or avoidance of complications associated with intravitreal injection of anti-VEGF therapy. Such complications may include, for example, one or more of the following: infective endophthalmitis, aseptic endophthalmitis, retinal vasculitis, and / or transient intraocular pressure (IOP) spikes. This may be a comparison, for example, between subjects treated according to a fifth aspect of the present invention and subjects treated with anti-VEGF therapy alone, or between subjects receiving anti-VEGF monotherapy who achieved the same preventive or therapeutic endpoint.

[0110] Optionally, according to a third, fourth, or fifth aspect of the present invention, the subject is typically a human subject such as a human male and / or human female. The subject may be an adult, or less typically, a young person, a child, or a newborn.

[0111] Optionally, according to a third, fourth, or fifth aspect of the present invention, the subject is a human adult and / or having a body weight of about 50 kg to about 150 kg. Where used herein, the term “about” may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0112] Optionally, according to a third, fourth, or fifth aspect of the present invention, the subject may be additionally or alternatively characterized by one or more of the following: a. The target group is people aged approximately 50 and over. b. The subjects are those who do not have or are not suspected of having cardiovascular disease (myocardial infarction, stroke, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis), or diseases related to cardiovascular disease (for example, sickle cell anemia, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (Behçet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably diabetes mellitus and diabetes mellitus vera). c. The subjects are those who do not have a history of cardiovascular disease (myocardial infarction, stroke, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis) or cardiovascular disease-related diseases (sickle cell anemia, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (e.g., Behçet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably excluding diabetes mellitus and diabetes mellitus vera), d. The subjects are those who do not have a family history of cardiovascular disease (myocardial infarction, stroke, atherosclerosis, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis) or a disease related to cardiovascular disease (e.g., sickle cell disease, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (e.g., Behçet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably excluding diabetes mellitus and diabetes mellitus vera), and / or e. The subject does not have, or is not suspected of having, one or more conditions selected from solid tumors, lymphomas and / or hematological malignancies, such as breast cancer, endometrial cancer, cervical cancer, ovarian cancer, lung cancer, head and / or cervical cancer, brain cancer, thyroid cancer, esophageal cancer, stomach cancer, colorectal and / or rectal cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, bone cancer, lymphoma (such as Hodgkin lymphoma and / or non-Hodgkin lymphoma), and / or hematological malignancies (such as acute lymphoblastic leukemia, acute myeloid leukemia, and / or chronic myeloid leukemia).

[0113] In one embodiment of the third, fourth, or fifth aspect of the present invention, a first dose of a molecule containing or comprising the annexin A5 protein (and / or any subsequent round of administration of the annexin A5 component) is administered to a subject at least once per day. With respect to any given round of administration, the dose of the annexin A5 molecule may be optionally administered to the subject at least once per day for at least two, three, four, five, six, or seven days within a period of seven days (one week) out of eight days, such as a period of one, two, three, four, or six or more consecutive days, and / or a period of at least one week, two, three, or four weeks or more.

[0114] In some embodiments of the third, fourth, or fifth aspects of the present invention, a molecule containing or comprising the annexin A5 protein, or a composition containing this molecule, is present in the same composition as the anti-VEGF agent.

[0115] In other embodiments of the third, fourth, or fifth aspects of the present invention, a molecule containing or comprising the annexin A5 protein, or a composition containing the molecule, is formulated separately from the anti-VEGF agent. In this case, the anti-VEGF and annexin A5 components can be administered to the subject simultaneously or sequentially.

[0116] In the case of sequential administration of annexin A5 component and anti-VEGF component, in some examples of the third, fourth, or fifth aspects of the present invention, the molecule containing or consisting of annexin A5 protein ("annexin A5 component") is administered to the subject as the first round of administration of annexin A5 component before the initiation of anti-VEGF therapy. In other examples, the molecule containing or consisting of annexin A5 protein may be administered to the subject as the first round of administration of annexin A5 component after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. For example, in the case of a form of anti-VEGF therapy (in the absence of annexin A5 co-therapy) that requires multiple rounds of anti-VEGF administration to achieve a prophylactic or therapeutic effect or a prophylactic or therapeutic endpoint in the subject, the annexin A5 component may be administered to the subject as the first round of administration of annexin A5 component before the initiation of anti-VEGF therapy, or after the initiation of anti-VEGF therapy, but before one or more subsequent rounds of administration of any of the anti-VEGF components.

[0117] In some embodiments of the third, fourth, and / or fifth aspects of the present invention, the subject may also receive a second round of administration of annexin A5 component. In some examples, annexin A5 component is administered to the subject as a first round of administration, followed by a second round of administration, before the initiation of anti-VEGF therapy. In other examples, annexin A5 component is administered to the subject as a first round of administration, before the initiation of anti-VEGF therapy, and then administered as a second round of administration, after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other examples, annexin A5 component is administered to the subject as a first round of administration, followed by a second round of administration, in which case both the first and second rounds of administration of annexin A5 component occur after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In some options, there is no round of anti-VEGF therapy between the first and second rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between the first and second rounds of annexin A5 administration. In other options, there may be two or more rounds of anti-VEGF therapy between the first and second rounds of annexin A5 administration.

[0118] In some embodiments of the third, fourth, and / or fifth aspects of the present invention, the subject may also receive a third round of administration of annexin A5 component. In some examples, annexin A5 component is administered to the subject as a first round of administration, followed by a second round of administration, and then a third round of administration, all of which are before the initiation of anti-VEGF therapy. In other examples, annexin A5 component is administered to the subject as a first round of administration, followed by a second round of administration, both of which are before the initiation of anti-VEGF therapy, and annexin A5 component is administered to the subject as a third round of administration, after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other cases, annexin A5 is administered to the subject as the first round of annexin A5 administration before the initiation of anti-VEGF therapy, then as the second round of annexin A5 administration, and subsequently as the third round of annexin A5 administration, in which case both the second and third rounds of administration occur after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. In other cases, annexin A5 is administered to the subject as the first round of annexin A5 administration, followed by the second round of annexin A5 administration, and subsequently as the third round of annexin A5 administration, in which case all three rounds of annexin A5 administration occur after the initiation of anti-VEGF therapy, but typically before the completion of anti-VEGF therapy. In some options, there is no round of anti-VEGF therapy between the first and second rounds of annexin A5 administration and / or between the second and third rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between the first and second rounds of annexin A5 administration and / or between the second and third rounds of annexin A5 administration.In other options, there may be two or more rounds of anti-VEGF therapy between the first and second rounds of administration of annexin A5 component and / or between the second and third rounds of administration of annexin A5 component.

[0119] While the administration of one, two, or three rounds of annexin A5 component is described above, it is understood that one or more further rounds of annexin A5 component administration may be used beyond the third round. In some cases, all rounds of annexin A5 component administration are performed before the initiation of anti-VEGF therapy. In other cases, the first, second, third, or any subsequent round of annexin A5 component administration may be performed before the initiation of anti-VEGF therapy, and subsequent rounds of one or more annexin A5 component administrations may be performed after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In other cases, the rounds of annexin A5 component administration are performed after the initiation of anti-VEGF therapy, but typically before the termination of anti-VEGF therapy. In some options, there are no rounds of anti-VEGF therapy between the first and second rounds of annexin A5 administration, between the second and third rounds of annexin A5 administration, between the third round and subsequent rounds of annexin A5 administration, and / or between any of the multiple subsequent rounds of annexin A5 administration. In other options, there may be a single round of anti-VEGF therapy between any one or more of the first and second rounds of annexin A5 administration, the second and third rounds of annexin A5 administration, the third round and subsequent rounds of annexin A5 administration, and / or between any of the multiple subsequent rounds of annexin A5 administration. In other options, there may be two or more rounds of anti-VEGF therapy between any one or more of the following: the first and second rounds of administration of annexin A5 component, the second and third rounds of administration of annexin A5 component, the third round and subsequent rounds of administration of annexin A5 component, and / or multiple subsequent rounds of administration of annexin A5 component.

[0120] As a result of administering one or more rounds of the annexin A5 component, it may be possible to reduce, or even completely avoid, the frequency and / or number of subsequent rounds of administration of the anti-VEGF component following the initial administration to the subject for a specified period, without substantially reducing the prophylactic or therapeutic efficacy of anti-VEGF therapy. This can be compared, for example, to a suitable control, such as a subject or cohort of subjects, who have never been treated with a molecule containing or consisting of the annexin A5 protein over the same specified period but have received multiple doses of the anti-VEGF component (e.g., consecutive doses approximately every 28 days (e.g., ±7, 6, 5, 4, 3, 2, or 1 day)). The specified period may be, for example, about, up to, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months, or about, up to, or at least 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “approximately” may refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period. The specified period may be until the end of the therapeutic or preventive need for the subjects. In some embodiments, a cohort may be two or more subjects, approximately, at least, or more than three, four, five, ten, fifteen, twenty, twenty-five, thirty, forty, fifty, sixty, seventy, eighty, ninety, or one hundred subjects.

[0121] In embodiments of the third, fourth, or fifth aspects of the present invention, a molecule comprising or consisting of annexin A5 protein may be administered to a subject before the initiation of anti-VEGF therapy in order to reduce the frequency of administration of the anti-VEGF component following the initial administration of the annexin A5 component. This can refer, for example, to the period between the initial administration of the annexin A5 component to the subject and the first administration of the anti-VEGF component to the subject, i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months or more, or a period exceeding 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years, provided that the subject receives at least one administration of the anti-VEGF component during treatment. In this context, the term “about” can refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0122] Additionally or alternatively, in subjects receiving co-therapy with Annexin A5 according to a third, fourth, or fifth aspect of the present invention, the reduction in the frequency of subsequent rounds of administration of the anti-VEGF component following the initial administration of the anti-VEGF component may refer, for example, to the period between the initial administration of the anti-VEGF component to the subject and the second administration of the anti-VEGF component to the subject, i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months or more, or a period exceeding 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “about” may refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0123] Additionally or alternatively, in subjects receiving co-therapy with Annexin A5 according to a third, fourth, or fifth aspect of the present invention, the reduction in the frequency of subsequent rounds of administration of the anti-VEGF component following the initial administration of the anti-VEGF component may refer, for example, to a period between the second round and / or any subsequent round of administration of the anti-VEGF component and the subsequent round of anti-VEGF administration, i.e., a period exceeding 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months, or a period exceeding 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. In this context, the term “about” may refer to a period of ±50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the stated period.

[0124] For example, according to the third, fourth, or fifth aspect of the present invention, after the initial administration of an anti-VEGF component to the subject, the subject receiving co-therapy with annexin A5, i. Within a 12-month period from the start of treatment with Annexin A5 molecule or composition, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose, ii. Within a period of 10 months from the start of treatment with Annexin A5 molecules or compositions, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 dose. iii. Within an 8-month period from the start of treatment with Annexin A5 molecule or composition, 8, 7, 6, 5, 4, 3, 2 or less than 1 dose, and / or iv. Six, five, four, three, two, or less than one dose of an anti-VEGF agent may be administered consecutively within a period of six months from the initiation of treatment with Annexin A5 molecule or composition.

[0125] In one embodiment of the third, fourth, or fifth aspect of the present invention, a molecule containing or comprising annexin A5 protein is administered to the target vascular system as a systemic administration, for example, by intravenous administration. When administered intravenously, the molecule containing or comprising annexin A5 protein, or a composition containing this molecule, may be used in the form of a sterile aqueous solution that can contain, for example, other substances, such as a salt or glucose sufficient to make the solution isotonic with blood. The sterile aqueous solution should be preferably buffered as needed (preferably to a pH of 3 to 9). For example, optionally, the aqueous solution is buffered at a pH within the range selected from approximately pH 4 to approximately pH 8, approximately pH 5 to approximately pH 8, approximately pH 6 to approximately pH 8, approximately pH 6.5 to approximately pH 7.9, approximately pH 6.8 to approximately pH 7.7, approximately pH 7.0 to approximately pH 7.5, approximately pH 7.1 to approximately pH 7.4, approximately pH 7.1 to approximately pH 7.4, and optionally, approximately pH 7.1, 7.2, 7.3, or 7.4, most preferably approximately pH 7.2. In this context, the term "approximately" when applied to the nearest integer pH value may refer to a value with a pH unit of ±0.5, 0.4, 0.3, 0.2, or 0.1 of the stated value, while the term "approximately" when applied to a pH value with one decimal place may refer to a value with a pH unit of ±0.05, 0.04, 0.03, 0.02, or 0.01 of the stated value.

[0126] In relation to third, fourth, or fifth aspects of the present invention, while these are disclosed herein as treatment options, it is not preferable to directly administer molecules containing or consisting of monomeric annexin A5 protein to the eye when treating ocular conditions such as macular edema, RVO, and / or conditions associated with macular edema and / or RVO. For example, the annexin A5 molecule may be administered by intravitreal injection. Alternatively, the annexin A5 molecule may be directly administered to the target retinal vein (such as the central retinal vein, retinal branch veins, and / or hemiretinal veins) by, for example, catheterization. However, this is preferable to avoid.

[0127] The annexin A5 protein used in the third, fourth, or fifth aspect of the present invention may be as described above for the first and / or second aspect of the present invention.

[0128] In some embodiments of the third, fourth, or fifth aspects of the present invention, the molecule comprising annexin A5 protein for use in the first and / or second aspects of the present invention may be a conjugate in which annexin A5 protein (such as monomeric annexin A5 protein) is conjugated to a portion such as a non-proteinoid portion. Non-limitingly, this portion may be, for example, a therapeutic, prophylactic, diagnostic, prognostic, or theragnostic portion. In some embodiments, this portion may be a drug (such as a non-proteinoid drug) and / or a radioactive portion (for example, if the conjugate is suitable for use in radioimmunotherapy ("RIT").

[0129] Furthermore, compositions comprising molecules containing or consisting of the annexin A5 protein described herein, for use according to a third, fourth, or fifth aspect of the present invention, are also provided herein. Such compositions may, for example, be unit dose compositions. A unit dose composition may contain, for example, a total dose of annexin A5 protein of about 5 mg, about 4 mg, about 3 mg, about 2 mg, or less than about 2 mg, such as about 0.1 to 1.5 mg, about 1 mg or less. In this context, the term "about" when applied to a unit dose value in mg described for the nearest integer may refer to a value that is ±0.5, 0.4, 0.3, 0.2, or 0.1 mg of the described value, while the term "about" when applied to a unit dose value in mg described to one decimal place may refer to a value that is ±0.05, 0.04, 0.03, 0.02, or 0.01 mg of the described value.

[0130] Optionally, the composition is a pharmaceutical composition, and further optionally, the composition further comprises one or more pharmaceutically acceptable carriers. In some embodiments, the composition comprises a molecule containing or comprising the annexin A5 protein and one or more pharmaceutically acceptable carriers. Optionally, in addition to use as a combination therapy with anti-VEGF agents, the composition is intended for use as a combination therapy with one or more other agents. Optionally, the annexin A5 molecule or its composition is used as a first-line treatment for a disease, disorder, or condition to be treated, with or without one or more other agents.

[0131] According to third, fourth, or fifth aspects of the present invention, in some embodiments, a molecule comprising or consisting of the annexin A5 protein or a composition comprising the molecule is intended for and / or used as a combination therapy with one or more other agents, and the subject is administered the annexin A5 molecule separately from the other agents in the same composition, simultaneously with or sequentially with a separate formulation containing the same.

[0132] Optionally, according to a third, fourth, or fifth aspect of the present invention, a molecule containing or comprising the annexin A5 protein and an anti-VEGF agent are administered together, either simultaneously in separate doses or sequentially in separate doses. In some embodiments, when the administrations are separate and sequential, the interval between consecutive administrations of the annexin A5 molecule and the anti-VEGF agent is about 28 days or less, such as about 3 weeks or less, about 2 weeks or less, about 1 week or less, about 6, 5, 4, 3, 2, or 1 day or less (e.g., about 24, 28, 12, 6, 5, 4, 3, 2, or 1 hour or less). However, studies reported in this application have demonstrated that administration of annexin A5 can delay the need for anti-VEGF therapy for a considerably longer period. Therefore, in other embodiments, when the administrations are separate and sequential, the interval between consecutive administrations of the annexin A5 molecule and the anti-VEGF agent is longer than about 28 days. For example, as stated above, Annexin A5 administration may be carried out over multiple days and may include multiple repeated administrations to the subject. For example, the Annexin A5 molecule may be administered to the subject for two or more consecutive days, such as three, four, five, six, seven or more consecutive days, and / or for at least two, three, five, five, six, or seven days within a period of seven days (one week) out of eight days, such as at least one week, two weeks, three weeks, four weeks or more. When Annexin A5 administration is carried out over multiple days, in some embodiments, the period between consecutive administrations of the Annexin A5 molecule and the anti-VEGF agent is calculated with respect to the start date of Annexin A5 administration. In other embodiments, the period is calculated with respect to the end date of Annexin A5 administration.

[0133] This application also provides, with reference to the foregoing, molecules comprising or consisting of the annexin A5 protein (such as a molecule comprising monomeric annexin A5) or compositions comprising such molecule, or methods of use or for use substantially described herein.

[0134] definition As used herein, the terms “molecule comprising or consisting of annexin A5 protein” and “molecule consisting of monomeric annexin A5 protein” are further defined in Section B of the following descriptions of embodiments for carrying out the present invention. It optionally further includes the meaning of a molecule comprising, essentially consisting of, or a composition comprising annexin A5 protein (such as a molecule consisting of monomeric annexin A5 protein). Such composition may, for example, be a pharmaceutically acceptable or veterinarily acceptable composition.

[0135] A "pharmaceutically acceptable" composition may be one that is safe for administration to a subject, such as a human subject, by injection, such as intravenous, subcutaneous, or intramuscular injection. Most preferably, the injection is intravenous.

[0136] A "veterinarily acceptable" composition may be one that is safe for administration to an animal subject by injection, such as intravenous, subcutaneous, or intramuscular injection. Most preferably, the injection is intravenous.

[0137] Accordingly, the pharmaceutical or veterinary compositions according to the present invention contain, essentially consist of, or may consist of, a molecule comprising monomeric annexin A5 protein mixed with a pharmaceutically or veterinarily acceptable adjuvant, diluent, and / or carrier, which is typically selected with respect to the intended route of administration and standard pharmacopoeia. The compositions may be in the form of immediate-release, delayed-release, or controlled-release applications. The formulations may be unit doses, e.g., a daily dose or unit of the active ingredient, less than a daily dose, or an appropriate proportion thereof.

[0138] The "intravitreous for sterile injection" composition may be a composition that is safe for administration to subjects, especially human subjects, by direct injection into the vitreous humor of the subject's eye.

[0139] Accordingly, the “intravitreal composition for sterile injection” according to the present invention comprises, essentially comprises, or may comprise a molecule consisting of monomeric annexin A5 protein mixed with an adjuvant, diluent, and / or carrier, selected for its suitability and / or tolerance when directly introduced into the vitreous humor of the eye. The composition may be in the form of immediate-release, delayed-release, or controlled-release applications. The formulation may be a unit dose, for example, a unit dose containing a daily dose or units of the active ingredient.

[0140] The terms “pharmaceutically acceptable,” “pharmacologically acceptable,” and “veterinary acceptable” refer to compositions that, when administered to animals, such as humans, as necessary, do not cause adverse, allergic, or other undesirable reactions. The same considerations apply to sterile intravitreous compositions for injection. The preparation of such compositions is known to those skilled in the art in light of this disclosure, as exemplified by Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990 (incorporated herein by reference). Furthermore, it will be understood that, for animal (e.g., human) administration, preparations should meet standards of sterility, pyrogenicity, general safety, and purity, such as those required by the FDA Office of Biological Standards.

[0141] As used herein, “pharmaceutically acceptable carrier” and “veterinarily acceptable carrier” include, as known to those skilled in the art, all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antimicrobial agents, antifungal agents), isotonic agents, salts, preservatives, drugs, drug stabilizers, excipients, disintegrants, similar substances, and combinations thereof. Unless any conventional carrier is incompatible with the active ingredient, its use in a therapeutic or pharmaceutical composition is intended.

[0142] Molecules containing or consisting of annexin A5 protein (such as molecules consisting of monomeric annexin A5 protein), or compositions containing such molecules (such as in mixture with one or more pharmaceutically acceptable carriers or veterinarily acceptable carriers), may be administered parenterally to a subject, for example, intravenously, intra-arterially, intraperitoneally, intraarachnoidally, intraventricularly, intrasternally, intracranially, intramuscularly or subcutaneously, or by infusion techniques.

[0143] One option is to avoid direct administration to the eye, such as intravitreal injection.

[0144] Alternatively, the molecule comprising monomeric annexin A5 protein may be administered directly to the eye. For example, in one optional and preferred embodiment, the molecule comprising monomeric annexin A5 protein may be administered to one or both of the target eyes by intravitreal injection. According to this embodiment, monomeric annexin A5 protein may be administered to the patient in the form of a sterile intravitreal composition for injection, for example, as defined according to a fourth aspect of the present invention.

[0145] Alternatively, a molecule containing or consisting of annexin A5 protein (such as a molecule consisting of monomeric annexin A5 protein) may be directly administered to one or both retinal veins of the target eye (such as the central retinal vein, retinal branch veins, and / or hemiretinal veins) by, for example, catheterization.

[0146] Systemic administration, such as intravenous administration, is a preferred embodiment of the first, second, third, fourth, and fifth aspects of the present invention. A molecule containing or consisting of annexin A5 protein (such as a molecule consisting of monomeric annexin A5 protein) or a composition containing such a molecule may be used in the form of a sterile aqueous solution that may contain, for example, other substances, such as a salt or glucose sufficient to make the solution isotonic with blood. The aqueous solution should be preferably buffered as needed (preferably to a pH of 3 to 9). For example, optionally, the aqueous solution may be buffered to a pH within the range selected from approximately pH 4 to approximately pH 8, approximately pH 5 to approximately pH 8, approximately pH 6 to approximately pH 8, approximately pH 6 to approximately pH 8, approximately pH 6.5 to approximately pH 7.9, approximately pH 6.8 to approximately pH 7.7, approximately pH 7.0 to approximately pH 7.5, approximately pH 7.1 to approximately pH 7.4, approximately pH 7.1 to approximately pH 7.4, and optionally, approximately pH 7.1, 7.2, 7.3, or 7.4, most preferably approximately pH 7.2. In this context, the term "approximately" when applied to pH values ​​stated as the nearest integer may refer to a value with a pH of ±0.5, 0.4, 0.3, 0.2, or 0.1 of the stated value, while the term "approximately" when applied to pH values ​​stated to one decimal place may refer to a value with a pH of ±0.05, 0.04, 0.03, 0.02, or 0.01 of the stated value. The preparation of suitable parenteral formulations under sterile conditions is readily achieved by standard pharmaceutical techniques well known to those skilled in the art.

[0147] Formulations suitable for parenteral administration typically include aqueous and non-aqueous sterile injection solutions that may contain antioxidants, buffers, bacteriostatic agents, and solutes that areotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions that may contain suspending agents and thickeners. Formulations may be in unit dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in a freeze-dried state requiring only the addition of a sterile liquid carrier, such as water for injection, immediately before use. Immediate injection solutions and suspensions may be prepared from the types of sterile powders, granules, and tablets previously described.

[0148] In one embodiment, a pharmaceutically acceptable or veterinarily acceptable composition (for example, one that may be used in any of the first, second, third, and / or fifth aspects of the present invention), and / or an intravitreous composition for sterile injection (for example, one defined by the fourth aspect of the present invention) may be defined as safe for injection if it does not contain or substantially contains endotoxin. Endotoxin is often used synonymously with the term lipopolysaccharide, which is the main component of the extracellular wall of Gram-negative bacteria. It consists of a polysaccharide (sugar) chain and a lipid portion known as lipid A, which is responsible for the toxic effects observed in endotoxin. The polysaccharide chain is highly variable between different bacteria, determining the serotype of the endotoxin, and the lipid component is also highly variable, so that a single endotoxin sample may contain tens to hundreds of different molecular species. Endotoxins are about 10 kDa in size, but can form large aggregates up to 1000 kDa. Endotoxins are typically harmful and pyrogenic in therapeutic compositions, and regulatory authorities impose strict limits on the permissible levels of endotoxins in pharmaceutical compositions. Therefore, the level of endotoxin in the compositions according to the present invention should be minimized, and may be less than 100 endotoxin units (EU) per dose, e.g., less than 90, 80, 70, 60, 50, 40, 30, 20, 10, 5, 4, 3, 2, or 1 EU / dose. The concentration of endotoxin in the compositions according to the present invention may be less than 200 EU / m3, e.g., 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, 5, 4, 3, 2, or 1 EU / m3. Methods for measuring endotoxin levels, such as the limulus amoebocyte assay (LAL), are well known in the art.

[0149] A composition containing or comprising a molecule comprising annexin A5 protein (such as a molecule comprising monomeric annexin A5 protein) may optionally contain the annexin A5 molecule as the sole activator. In other words, in one embodiment, the composition (or pharmaceutical composition) may consist of a molecule containing or comprising annexin A5 protein (such as a molecule comprising monomeric annexin A5 protein) and one or more pharmaceutically acceptable carriers or one or more veterinarily acceptable carriers. For example, compositions in which annexin A5 is formulated with, conjugated with, or expressed as a fusion protein with one or more further proteins or non-protein agents that can provide an additional therapeutic effect may be optionally excluded. This additional therapeutic effect may be either a condition of macular edema and / or retinal vein occlusion, or a condition of therapeutic effect unrelated to the treatment of macular edema and / or retinal vein occlusion. To avoid misunderstanding, the molecule consisting of monomeric annexin A5 protein is not dimeric annexin A5 (commonly referred to as "dianexin" in the art) nor is it PEGylated annexin A5.

[0150] Other definitions: The term “treatment” is as understood by those skilled in the art of medicine. As used herein, the term “treatment” may include any treatment of a subject, in particular a condition in a human or other mammal (for example, a condition in which the condition is macular edema, retinal vein occlusion, or a condition associated with macular edema or retinal vein occlusion), and the following effects: (i) To inhibit a condition (for example, macular edema or retinal vein occlusion, and / or other diseases, disorders, or conditions related to and / or caused by macular edema or RVO), i.e., to delay, reduce, or cessate the onset of the condition. (ii) To alleviate a condition (for example, macular edema or retinal vein occlusion, and / or other diseases, disorders, or conditions related to and / or caused by macular edema or RVO), that is, to cause regression of the condition in an object having the condition, or (iii) To cure a condition (for example, macular edema or retinal vein occlusion, and / or other diseases, disorders, or conditions related to and / or caused by macular edema or RVO), that is, to restore an object having a condition to a state of health in which the condition is no longer detectable. It must include one or more of the following, at your discretion.

[0151] The term “prevention” is as understood by those skilled in the art of medicine. As used herein, the term “prevention” may include any preventive treatment of a condition in a subject, in particular in humans or other mammals (for example, a condition in which the condition is related to macular edema, retinal vein occlusion, macular edema, or RVO), and the following effects: (i) To prevent a condition (e.g., macular edema or retinal vein occlusion, and / or other diseases, disorders, or conditions associated with and / or caused by macular edema or RVO) from occurring in a subject (e.g., a subject who is predisposed to developing the condition or at risk of developing it, but has not yet been diagnosed as having it), that is, to stop the subject from developing the condition. (ii) Delaying the onset of a condition in the subject (for example, macular edema or retinal vein occlusion, and / or other diseases, disorders, or conditions related to and / or caused by macular edema or RVO), that is, delaying the onset of the condition in the subject until later in the subject's life. (iii) Limiting the occurrence of a condition in the subject (e.g., macular edema or retinal vein occlusion, and / or other diseases, disorders, or conditions related to and / or caused by macular edema or RVO), for example, reducing the degree to which the subject suffers from the condition, or (iv) To prevent one or more symptoms of a condition in the subject (e.g., macular edema or retinal vein occlusion, and / or other diseases, disorders, or conditions related to and / or caused by macular edema or RVO), i.e., to stop the subject from developing one or more symptoms of the condition. It must include one or more of the following, at your discretion.

[0152] For the prevention of other diseases, disorders, or conditions related to and / or caused by macular edema or RVO, subjects may be, on a voluntary basis, individuals who already have, are suspected of having, or have been diagnosed with macular edema or RVO.

[0153] Terms such as “optionally,” “preferably,” “etc.,” and “for example” mean that the features, characteristics, events, or circumstances described thereafter may or may not occur, and that the description includes both cases in which such features, characteristics, events, or circumstances occur and cases in which they do not. To avoid misunderstanding, please understand that in some embodiments of the present invention, any features, characteristics, events, or circumstances described in the context of “optionally,” “preferably,” “etc.,” or “for example” may be excluded from the scope of the claims.

[0154] As used herein, the term “subject” includes any living human or animal to which the treatment of the present invention is applied. Exemplary subjects, though not limited to those described herein, are further defined in Section C of the following descriptions of embodiments for carrying out the invention. More preferably, the subject is a human subject.

[0155] As used herein, the term “elevated” means that a substance (e.g., a cell type such as red blood cells (i.e., erythrocytes), or a cellular marker such as an annexin A5 binding site on red blood cells) is present in a greater amount in subjects with macular edema and / or retinal vein occlusion compared to the amount present on average in subjects or groups of subjects (e.g., at least 10,000 or 1,000 subjects) without macular edema and / or retinal vein occlusion.

[0156] As used herein, the term “distal” means that the administration of a molecule consisting of monomeric annexin A5 protein is not an intraocular or direct administration to the eye diagnosed with macular edema and / or retinal vein occlusion.

[0157] The term "therapeutic effective dose" refers to the amount of a drug, compound, protein (e.g., a molecule containing or comprising annexin A5 protein as defined herein) or pharmaceutical composition sufficient to produce a beneficial effect or desirable outcome, including clinical outcomes related to the treatment or prevention of macular edema, retinal vein occlusion, or diseases, disorders, or conditions associated with and / or caused by macular edema and / or retinal vein occlusion. The effective dose may be administered in one or more doses. For the purposes of the present invention, the effective dose of a drug, compound, protein (e.g., a molecule containing or comprising annexin A5 protein as defined herein), pharmaceutical composition or veterinary composition is sufficient to treat, alleviate, reduce the severity of, and / or prevent macular edema and / or retinal vein occlusion, and / or other diseases, disorders, or conditions associated with and / or caused by macular edema and / or retinal vein occlusion. As understood in a clinical context, an effective dose of a drug, compound, protein (e.g., a molecule containing or comprising the annexin A5 protein as defined herein) or pharmaceutical composition may or may not be achieved when administered in combination (separately, sequentially, or concurrently) with other drugs, compounds, proteins, or pharmaceuticals, such as those described elsewhere in this application. Therefore, “effective dose” may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered given in an effective dose if, in combination with one or more other agents, a desired outcome can be produced or achieved. [Brief explanation of the drawing]

[0158] [Figure 1]This panel shows the results of a phosphatidylserine (PS) assay in RBCs derived from patients with RVO. This flow cytometry in vitro assay uses FITC annexin A5 ("ANXV") and PE anti-CD235a as positive markers for RBCs. Panel A: Ca2+ is absent in the negative control, while Ca2+ is present in all other samples, enabling in vitro binding of FITC annexin A5. Panel B: PS exogenization to RBCs at screening, imaging date, and follow-up time. [Figure 2] This image shows fluorescein angiography (FA) performed using fluorescein administered intravenously prior to systemic administration of a fixed dose of 1 mg of ANXV-800CW. Red arrows indicate occlusion. Dark areas marked by red ellipses correspond to areas of injury with hemorrhage, indicating the presence of ischemia. [Figure 3] The near-infrared (NIR) fluorescence image (Panel B) was created t=5 minutes after a systemic injection of 1.0 mg of ANXV-800CW. The NIR image at t=229 minutes (Panel A) is shown. Red arrows indicate the occlusion site, and yellow arrows mark the vessels anterior to the occlusion site in the injured area where bleeding is present. Light blue arrows indicate large veins within the optic nerve. The dark area enclosed by the red oval corresponds to the injured area where bleeding is present. This oval marks approximately the same anatomical location as the red oval in Figure 2. The settings used for the Heidelberg Spectralis camera system are similar in Panels A and B. [Figure 4](A) This shows a retinal sensitivity map obtained by microperimetry (MMP) of treatment-naïve RVO patients (Target F) treated with ANXV therapy as first-line therapy. These findings demonstrate that in patients who did not receive anti-VEGF therapy during the study period, there was a clear improvement at 15 days post-treatment and complete recovery by 120 days post-treatment. (B) This shows the results of fluorescein angiography (FA) imaging of blood flow in the retinal vascular system for Target F during the same study period, demonstrating that with ANXV therapy, there was a clear improvement at 15 days post-treatment and complete recovery by 29 or 120 days post-treatment. [Figure 5] This shows optical coherence tomography (OCT) measurements of macular retinal thickness in treatment-naive RVO patients (Control A) who were followed over a 16-month study period, treated with ANXV therapy as first-line therapy at the start of the study, and received a single anti-VEGF injection around day 32 of the study. At 16 months, despite receiving only a single anti-VEGF treatment during the study period, visual acuity remained stable at 20 / 60. [Figure 6] The application of TNFα resulted in a decrease in cell viability compared to control cells without TNFα (Column 1) (Column 2), whereas the application of annexin A5 in addition to TNFα inhibited cell death (Column 3) and resulted in cell viability at a level comparable to that observed in the control group. [Figure 7]Treatment with annexin A5 protein increases cell viability in a dose-dependent manner. The application of IL1β resulted in decreased cell viability. This is shown in the figure as the flow cytometry curve for samples treated with IL1β alone (marked in the figure) is shifted to be the curve furthest from the y-axis, while the control cells not treated with IL1β are shown as being in the center of the graph. This specific position of these curves indicates that cell death occurs at a higher level when IL1β is applied. Since the control curve and the annexin A5 protein curves for all three doses are in the same position, it is also shown that applying annexin A5 protein alone at all three doses does not cause increased cell death compared to the control sample. Finally, the application of annexin A5 in addition to IL1β inhibits cell death in a dose-dependent manner, with doses of 20 μg and 5 μg showing the same levels of cell viability observed in the control. This is shown by the fact that the flow cytometry curves for annexin A5 at doses of 20 μg and 5 μg are in the center of the graph (marked in the figure), coinciding with the control curve and the annexin A5 protein alone curve. The 100 ng dose also suppresses cell death, but to a lesser extent, which can be seen from the fact that the 100 ng flow cytometry curve (marked in the figure) is between the center of the graph and the IL1-β alone curve. [Figure 8] This study demonstrates that ICAM1 expression decreases with treatment with Annexin A5. Experiments were performed on cell cultures using flow cytometry. TNFα application increased ICAM1 surface expression compared to control cells without TNFα application (Column 1) (Column 2). Application of Annexin A5 in addition to TNFα decreased ICAM1 surface expression (Column 3). [Figure 9]This study demonstrates that IL-6 expression decreases in a dose-dependent manner with treatment using Annexin A5. Application of IL-1β resulted in increased IL-6 expression compared to a control sample without IL-1β (Column 1) (Column 5). Application of Annexin A5 protein alone at all doses did not increase IL-6 expression (Columns 1-4). Application of Annexin A5 in addition to IL-1β resulted in a dose-dependent decrease in IL-6 gene expression (Columns 5-8). [Modes for carrying out the invention]

[0159] The established standard of treatment for macular edema and / or RVO and several other retinal diseases is by direct delivery of therapeutic agents (such as anti-VEGF agents) to the eye, via intravitreal injection (Haydinger et al, 2023 (cited above), Ip & Hendrick, 2018 (cited above), Cox et al., 2021, J. Clin. Med., 10(5):981). Since the retina is located at the back of the eye and surrounded by several protective layers, including the cornea, iris, and lens, such drugs are injected directly into the eye. These layers can make it difficult for drugs to reach the retina if administered by other routes. When a drug is injected into the eye, it is delivered directly to the retina and can act locally to target the cause of the problem. In part, the present invention is based on the remarkable finding that when annexin A5 protein is administered systemically (such as intravenously) to a subject, it can preferentially localize to the occlusion site in subjects with retinal vein occlusion (RVO), thereby providing a therapeutic effect in the treatment of RVO.

[0160] Furthermore, annexin A5 was found to possess biological activity that modulates various factors identified as contributing to dysregulation of the blood-retinal barrier (BRB) in subjects with macular edema and / or RVO, and to other therapeutic benefits in subjects with macular edema and / or RVO. The inventors also found that prevention and / or treatment of macular edema, retinal vein occlusion, and / or other diseases, disorders, or conditions associated with and / or caused by macular edema and / or retinal vein occlusion can be achieved by annexin A5 (including, but not limited to, monomeric annexin A5 protein) within a range of doses.

[0161] Even more remarkable is the finding that Annexin A5 therapy can substantially reduce the burden of downstream anti-VEGF therapy. Anti-VEGF therapy is well-established in the treatment of various ocular conditions, including macular edema, RVO, and related conditions.

[0162] Therefore, the present invention provides for the use of annexin A5 protein in combination therapy with anti-VEGF in the context of preventing or treating conditions such as macular edema, RVO, and conditions associated with each of these.

[0163] A. Macular edema Macular edema (also known as macular edema) is a pathological accumulation of fluid in the macula, the central region of the retina essential for high visual acuity. It is a common complication of numerous eye diseases, including diabetic retinopathy, retinal vascular occlusion, uveitis, post-cataract surgery inflammation (pseudophakic macular edema or Erving-Gas syndrome), retinal dystrophy, drug reactions, intraocular tumors, serous central chorioretinopathy, radiation retinopathy, and other retinal vascular abnormalities, including retinal microaneurysms and retinal telangiectasia (Tranos et al., 2004, Surv Ophthalmol, 49:470-90). Subretinal neovascularization observed in conditions such as age-related macular degeneration (AMD) is also associated with intraretinal and subretinal fluid accumulation (Trichonas et al., 2014, Br J Ophthalmol. 98:ii24).

[0164] Symptoms of macular edema include metamorphopsia, micropsia, blurred vision, central scotoma, and decreased contrast or color sensitivity (Haydinger et al., 2023).

[0165] Optionally, treatment of macular edema or related conditions is initiated, according to various embodiments of the present invention, within 10, 20, 30, 40, 50, or 60 minutes, within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or within 1, 2, 3, 4, 5, 6, or 7 days of the onset of one or more symptoms of macular edema. For example, combination therapy may be initiated by the first administration of the annexin A5 component of the cotherapy, or by the first administration of the anti-VEGF component of the cotherapy, or by the simultaneous first administration of both components, either as separate compositions or in combination in the same formulation.

[0166] The clinical diagnosis of macular edema can be difficult in mild cases or when it is impaired by pupillary dilation, cataracts, and other intermediate transparent media that impair fundus visibility. Therefore, fluorescein angiography and optical coherence tomography (OCT) are commonly used for diagnosis. The latter is preferred due to its non-invasive nature, accurate measurement of retinal thickness, and ability to identify other structural abnormalities such as epiretinal membranes and vitreomacular traction, and is the standard method for evaluating macular edema in clinical trials.

[0167] Therefore, in a preferred embodiment, a subject diagnosed with macular edema is a subject in whom macular edema is diagnosed by fluorescein angiography and / or optical coherence tomography.

[0168] Optionally, according to the present invention, treatment of macular edema or a related condition in a subject is initiated within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, or 60 minutes of the diagnosis of macular edema, within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or within 1, 2, 3, 4, 5, 6, or 7 days. For example, combination therapy may be initiated by the first administration of the annexin A5 component of the cotherapy, or by the first administration of the anti-VEGF component of the cotherapy, or by the simultaneous first administration of both components, either as separate compositions or in combination in the same formulation.

[0169] Retinal thickness is generally measured between the internal limiting membrane and Bruch's membrane. According to current spectral domain OCT technology, the mean macular thickness and central (foveal) macular thickness in a normal adult eye are 334 and 226 μm, respectively, and are thicker in males than in females. Three patterns of macular edema, cystoid macular edema, diffuse macular edema, and subretinal detachment, have been described using OCT. Diffuse retinal thickening is the main pattern in diabetic macular edema, while subretinal fluid accumulation is common in retinal vascular occlusion. In uveitis, cystoid macular edema and diffuse macular edema are typical forms of macular edema, while subretinal detachment is usually found in less than one-third of cases, often in combination with cystoid or diffuse edema. In some embodiments of the present invention, macular edema is selected from the group consisting of one or more of cystoid macular edema, diffuse macular edema, and subretinal detachment.

[0170] Distortion of retinal structures caused by fluid accumulation in the macula can lead to vision loss, and if the condition becomes chronic, it can result in irreversible visual impairment due to scarring. Macular edema is a leading cause of vision loss in patients with diabetes mellitus, retinal vein occlusion, and uveitis. Population studies have shown that the mean prevalence of diabetic macular edema is about 6% in type 1 diabetes and slightly higher in type 2 diabetes. Macular edema is extremely common in central retinal vein occlusion, but in branch retinal vein occlusion, macular edema occurs in 30% of cases. About one-third of patients with uveitis develop macular edema, more frequently in intermediate uveitis and panuveitis, and less common in anterior uveitis. The burden of macular edema is related not only to the reduced quality of life experienced by patients, but also to the economic costs associated with the chronicity of the condition and frequent visits to treatment and medical centers. In particular, in uveitis-induced macular edema, the most affected group is the working-age population, and the socioeconomic impact of this condition is increasing.

[0171] In some embodiments of the present invention, subjects who have, have been diagnosed with, or are suspected of having macular edema may be subjects who have, have been diagnosed with, or are suspected of having one or more conditions selected from diabetes mellitus, diabetic retinopathy, retinal vein occlusion (e.g., CRVO, BRVO and / or HRVO), and / or uveitis (e.g., intermediate uveitis and / or panuveitis).

[0172] In some embodiments of the present invention, subjects who have, have been diagnosed with, or are suspected of having macular edema may be subjects who have received, are receiving, or are scheduled to receive a treatment selected from the group consisting of macular edema-related therapies, e.g., anti-VEGF therapy (e.g., anti-VEGF therapy delivered to the eye by intravitreal injection), corticosteroids, NSAIDs, transsquamative vitrectomy, retinal laser photocoagulation, or anti-VEGF therapy enhanced with intravitreal corticosteroids and / or retinal laser photocoagulation.

[0173] An exemplary anti-VEGF therapy includes one of the following: • Ranibizumab (an FDA-approved anti-VEGF drug, which is a recombinant humanized antibody fragment against VEGF), • Aflibercept (another FDA-approved anti-VEGF drug, a recombinant protein containing ligand-binding domains of VEGF receptors 1 and 2 fused to the Fc portion of immunoglobulin G1), • Bevacizumab (a humanized monoclonal antibody against VEGF, widely used off-label) (Haydinger et al, 2023 (see above), Ip & Hendrick, 2018 (see above)), and / or Brolucizumab and / or falisimab (a newer anti-VEGF drug).

[0174] Corticosteroids are commonly used to manage macular edema, particularly in patients with non-infectious uveitis. Exemplary corticosteroids include difluprednate, triamcinolone acetonide, and dexamethasone, which can be used as monotherapy or in combination with systemic immunomodulatory therapy to achieve more rapid resolution of edema. The most common complications of topical corticosteroid treatment are elevated intraocular pressure and cataracts. Systemic use of corticosteroids is primarily indicated for bilateral macular edema and should be used for limited periods due to the wide range of side effects (Foster et al., 2016, Surv Ophthalmol, 61:1-17).

[0175] Non-steroidal anti-inflammatory drugs (NSAIDs) play an adjunct role in treatment and are primarily used for pseudophakic macular edema. Acetazolamide, a carbonic anhydrase (CA) inhibitor, may be used when corticosteroid therapy fails, but it is also sometimes used as a first-line treatment for pseudophakic macular edema or macular edema associated with retinal dystrophy.

[0176] Transsquamative vitrectomy is primarily used to treat organic factors contributing to macular edema, such as epiretinal membranes and vitreoretinal traction. It is a last resort for treating macular edema without organic causes, due to its inherent risks, including retinal damage and detachment, vitreous hemorrhage, cataracts, glaucoma, and ocular hypotension, as well as limited data on outcomes.

[0177] The fluid can accumulate in the center of the retina, or within cysts usually localized in the inner granular layer or the layer of Henle, or scattered throughout the subretinal space, potentially destroying and distorting retinal structures and impairing vision. Combined with the underlying pathology, edema can progress to irreversible tissue damage involving retinal cell death and permanent vision impairment. A common feature of most diseases that cause macular edema is dysfunction of the blood-retinal barrier (BRB), a set of structures that, in a healthy state, tightly regulate the passage of proteins, salts, metabolites, and other solutes between the blood and retinal tissue. Vasoactive growth factors and inflammatory cytokines, including vascular endothelial growth factor-A (VEGF) and tumor necrosis factor (TNF)-α, are upregulated in the ocular fluid in macular edema and are involved in BRB disruption. Dysregulation of solute inflow and accumulation in the retina disrupts the osmotic and hydrostatic balance, leading to fluid inflow when mechanisms maintaining fluid homeostasis are breached. Current treatments target proteins and processes involved in angiogenesis, inflammation, and blood-retinal barrier (BRB) dysfunction. This review provides an overview of the clinical aspects of macular edema and discusses the cellular and molecular mechanisms targeted by current treatments.

[0178] Conditions associated with macular edema include, but are not limited to, one or more of the following: dysregulation of the blood-retinal barrier (BRB), e.g., dysregulation of the medial and / or lateral BRB; dysfunction of the barrier between the vitreous and ciliary blood vessels; macular edema-related eye diseases, e.g., diabetic retinopathy, retinal vascular occlusion, uveitis, post-cataract surgery inflammation (pseudophakic macular edema or Irving-Gas syndrome); retinal dystrophy; drug reactions; intraocular tumors; serous central chorioretinopathy; radiation retinopathy; and other retinal vascular abnormalities, including retinal microaneurysms and retinal telangiectasia; changes in the subretinal neovascular membrane observed in conditions such as age-related macular degeneration (AMD); metamorphopsia; micropsia; blurred vision; central scotoma; and decreased contrast or color sensitivity.

[0179] Mechanism of macular edema Macular edema is a result of a failure of the mechanisms that maintain fluid homeostasis in the macular region (Haydinger et al, 2023 (see above)). The movement of fluid through capillaries throughout the body is classically explained by Stirling forces. The hydrostatic pressure gradient between the intravascular space and the extracellular space of the tissue tends to drive fluid into the tissue, which is counteracted by the colloidal osmotic pressure gradient that tends to guide fluid from the extracellular space into the blood vessels. A recent review by Cunha-Vaz discusses macular edema formation in relation to classical Stirling forces (Ophthalmologica, 2017, 237:1-10). The relative increase in hydrostatic pressure of blood vessels relative to retinal tissue, and the relative increase in colloidal osmotic pressure in retinal tissue relative to blood vessels are two forces that drive edema. Of these, macular edema in most diseases is thought to be caused by an increase in tissue colloid osmotic pressure resulting from the abnormal accumulation of solutes in the extracellular space of the retina (Haydinger et al, 2023 (see above)). The docile nature of retinal tissue, which can expand without obstruction into the vitreous humor, makes it particularly susceptible to edema, as an increase in tissue colloid osmotic pressure easily leads to an increase in tissue volume.

[0180] Dysregulation of solute inflow into the retina results from disruption of the blood-retinal barrier (BRB), which has been the main focus of much of the effort to understand the mechanism of this condition (Haydinger et al, 2023 (see above)).

[0181] The blood-retinal barrier (BRB) is a group of intraocular structures that separate blood from the neuroretina and extracellular space of the retina. It consists of two components: the medial BRB and the lateral BRB. The medial BRB is formed primarily by retinal endothelial cells (RECs) that surround the retinal blood vessels, while the lateral BRB is formed at the level of retinal pigment epithelial (RPE) cells. Since non-selective bidirectional diffusion of solutes can exist between the extracellular space of the retina and the vitreous humor, the barrier process between the vitreous humor and blood vessels in the ciliary body may also be considered to form part of the BRB; however, this region is not classified as either the medial or lateral BRB.

[0182] The walls of retinal vessels form an internal BRB. The luminal surface of retinal vessels is continuously covered by a squamous epithelial layer of RECs, and the membranes of adjacent RECs are held together by a continuous network of tight junctions. RECs are not fenestrated and contain few vesicles. RECs and their junctions form a barrier that facilitates the controlled passage of solutes between the bloodstream and the retina (Cunha-Vaz et al., 1966, Br J Ophthalmol., 50:441-53). The basement membrane completely covers the anti-luminal surface of RECs. Evidence collected in vivo indicates that the basement membrane does not directly constitute a barrier, but some evidence suggests that it may interfere with the diffusion of molecular tracers in vitro. Larger vessels have additional layers of smooth muscle, basement membrane, and adventitia, which vary in structure depending on the size and type of vessel.

[0183] The lateral BRB is not located in the vascular wall, but rather is formed by a monolayer of RPE cells located behind the retinal pigment epithelium, photoreceptors, and just opposite Bruch's membrane, which separates it from the vascular bed of the choroidal capillary plate (Haydinger et al, 2023 (see above)). In contrast to the lateral BRB, the choroidal vessels have thin, highly fenestrated endothelial cells and do not form a selective barrier. Thus, the epithelium is involved in maintaining the controlled environment necessary for retinal function.

[0184] Dysfunction of the blood-receptor ridge (BRB) underlies the development of macular edema (Haydinger et al, 2023 (see above)). Loss of selectivity for molecular exchange between blood and retinal tissue at either the medial or lateral BRB level allows for the entry of proteins and other solutes into the retinal tissue. Leakage is usually evident well before macular edema formation. Edema is likely to occur when the severity of leakage exceeds the solute and fluid clearance mechanisms. The clearance rate of solutes from the subretinal space is inversely proportional to their size. Injection of various sizes of tagged dextran tracers into patients with uveitis macular edema showed a leakage rate approximately proportional to the tracer size. Tracers up to 20 kDa leaked into the macula, while 150 kDa did not (Atkinson et al., 1991, Eye, 5:440-6). These data suggest that while greater solute infiltration is likely to present a greater challenge to clearance, complete BRB degradation, which allows for non-selective invasion of the maximum solute, is not necessarily required for edema formation.

[0185] This evidence suggests that several inflammatory cytokines act to disrupt BRB integrity (Haydinger et al, 2023 (see above), Park et al. 2019, J Diabetes Res, 2019:813741, Noma et al., 2020, J. Clin. Med, 9(1):3457).

[0186] VEGF plays a major role in driving the progression of retinal diseases and macular edema, a fact undeniably demonstrated by the widespread efficacy of anti-VEGF therapy (Haydinger et al., 2023 (see above), Park et al. 2019 (see above), Noma et al., 2020 (see above)).

[0187] Intravitreal levels of VEGF and IL-6 correlate with the severity of macular edema and the size of the non-perfusion area (Noma et al., 2009, Ophthalmology, 116:87-93; Noma et al, 2006, Graefes Arch Clin Exp Ophthalmol., 244:309-15). Evidence indicates that IL-6 is involved in diseases that lead to macular edema (Haydinger et al, 2023 (see above)). IL-6 is elevated in patients with uveitis (van Kooij et al., 2006, Am J Ophthalmol. 142:192-4). IL-6 levels are elevated in diabetic macular edema (Funatsu et al., 2002, Am J Ophthalmol. 133:70-7) and correlate with macular thickness (Oh et al., 2010, Curr Eye Res. 35:1116-27). IL-6 is also upregulated in patients with central retinal vein occlusion, particularly those with ischemia (Noma et al., 2009, Ophthalmology. 116:87-93). Its expression often correlates with VEGF expression (Haydinger et al, 2023 (see above)). Furthermore, it has been suggested that VEGF increases vascular permeability and further increases the expression of inflammatory cytokines, including MCP-1 and ICAM-1, which destroy BRBs, thereby causing the onset and progression of macular edema (Noma et al., 2020 (see above)).

[0188] Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) are also involved in the mechanism of BRB dysfunction in macular edema (Haydinger et al, 2023 (see above)).

[0189] TNF-α is a major inflammatory cytokine that has been shown to increase barrier permeability of the REC monolayer in vitro (Haydinger et al, 2023 (see above), Aveleira et al., 2010, Diabetes. 59:2872-82). When injected into the vitreous humor of rabbits, TNF-α increased the percentage of REC tight junctions that appeared open, and albumin was detected and leaked within these junctions (Luna et al., 1997, J Neurosci Res. 49:268-80). Larger vesicles also stained positively for albumin in the REC of treated eyes, but not in controls. These results indicate disruption of both transcellular and paracellular pathways. Interestingly, 24 hours after injection, the TNF-α-induced tight junction changes reversed, which is a promising sign for a potential therapeutic target. When injected into the vitreous humor of rats, it also induced barrier leakage, and the effect of the injection was recovered after 7 days (Bamforth et al., 1996, Acta Neuropathol. 91:624-32). Huang et al. evaluated the pathological role of TNF-α using two mouse models of diabetes (Ophthalmol Vis Sci. (2011) 52:1336-44), and found that vascular leakage in the later stages of the disease was completely prevented by TNF-α knockout.

[0190] Along with TNF-α, another major inflammatory cytokine of growing interest is IL-1β. Similar to TNF-α, when injected into the vitreous humor of rabbits, IL-1β increases the number of open, leaky tight junctions (140). IL-1β also increases the permeability of the REC monolayer in vitro (Aveleira et al., 2010 (see above)). In streptozotocin-diabetic rats, retinal IL-1β expression increased between 1.5 and 4.5 months of diabetes, and IL-1β mRNA was induced in vitro in bovine REC by high glucose (Liu et al., 2012, PLoS One.7:e36949), which then induces its own expression in vitro in REC, Müller cells, and brain astrocytic cells, which is inhibited by inhibition of protein kinase C.

[0191] B. Retinal vein occlusion Types of retinal vein occlusion Aspects of the present invention relate to the prevention and treatment of one or more diseases, disorders, or conditions related to and / or caused by retinal vein occlusion.

[0192] Retinal vein occlusion (sometimes called eye stroke) is a common retinal vascular disorder and one of the most common causes of vision loss worldwide. Generally, retinal vein occlusion is thought to be caused by the occlusion (i.e., blockage) of the retinal venous system due to thrombus formation or other factors. Other possible causes of retinal vein occlusion are thought to be vein occlusion caused by external pressure on the retinal veins, or diseases of the retinal vein wall due to vasculitis or other factors (see "Retinal Vein Occlusion (RVO) Guidelines" above).

[0193] Red blood cells can coagulate and form thrombi, a process known to be mediated by phosphatidylserine exposed on the surface of red blood cells. While not bound by theory, the applicant has characterized the role of annexin A5 in masking (or, in other words, covering or shielding) these phosphatidylserine on the surface of red blood cells as being able to avoid and treat thrombus formation in retinal vein occlusion. By characterizing the role of exposed phosphatidylserine in thrombus formation in retinal vein occlusion, the applicant has confirmed that it is possible to treat patients with substantially less monomeric annexin A5 than previously considered effective in the art.

[0194] In one embodiment, retinal vein occlusion is caused by occlusion of a blood vessel. For example, retinal vein occlusion may be associated with one or more of the following: the formation of one or more thrombi, observably enhanced hemagglutination and / or blood hyperviscosity, external pressure on the vein, and disease of the vein wall (e.g., vasculitis). Most preferably, retinal vein occlusion is associated with the formation of thrombi.

[0195] "Related" includes the retinal vein occlusion being caused by a particular mechanism (such as thrombus formation), and / or being suspected to be caused by a particular mechanism, and / or being diagnosed as being caused by a particular mechanism. As discussed herein, methods for identifying whether retinal vein occlusion is related to a particular mechanism will be known to those skilled in the art of medicine.

[0196] As is well known to those skilled in the art of medicine, thrombosis is generally considered to be a blood clot formed within a blood vessel (such as a vein) by the aggregation of platelets and / or red blood cells, which obstructs blood flow. Therefore, "thrombus" includes blood clots formed within blood vessels (such as veins) that obstruct blood flow.

[0197] Subjects may be classified as having observably enhanced hemagglutination and / or blood hyperviscosity by any appropriate method. This enhancement is compared to healthy subjects or to the mean for a group of healthy subjects (e.g., a group of 10, 50, or 100 healthy subjects). For example, the level of hemagglutination and / or blood hyperviscosity may be measured by determining the level of red blood cell adhesion to endothelial cells. A preferred methodology is described, for example, in Wauter et al, 2011, J. Thrombosis and Hemostasis, 9:1049-1055, which is incorporated herein by reference for the purpose of describing the red blood cell adhesion method.

[0198] More specifically, in one option, the level of erythrocyte adhesion to endothelial cells can be determined under static conditions. This may involve using radiometric techniques, such as those described, for example, in Kuypers et al, 1996, Blood, 87:1179-87 (the contents of which are incorporated herein by reference). Human microvascular endothelial cells (HMEC-1) may be cultured, and washed erythrocytes may be resuspended (e.g., to 25% hematocrit in Hanks Balanced Salt Solution, HBSS), preferably labeled (e.g., 51 Red blood cells were incubated with HMEC (e.g., 37°C for 30 minutes) after adding 0.5% human serum albumin (with chromium). Non-adherent red blood cells were removed by washing, and the remaining adhered red blood cells could be detected. Such detection may include, for example, lysing the remaining adhered red blood cells (e.g., with distilled water) and measuring the radioactivity in the lysate. Using such techniques, adhesion can be measured, for example, 1 mm. 2 This can be calculated as the number of red blood cells attached to the target area.

[0199] Under such static conditions, for example, in a healthy individual, typically 1 mm 2 It may show an attachment of 3700±300 red blood cells per 1 mm², but more generally, patients with CRVO have 1 mm².2 It may show 13,500 ± 700 red blood cells per unit. The subject to be treated according to the present invention is optionally determined, for example by the technique described above, and when evaluated for adhesion to endothelial cells under static conditions, 1 mm 2 A blood cell can be classified as having more than 4,000 red blood cells per 1 mm, for example, approximately 4,500, 5,000, 5,500, 6,000, 6,500, 7,000, 7,500, 8,000, 8,500, 9,000, 9,500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, or more. In this context, the term "approximately" refers to the value per 1 mm. 2 This refers to the number of red blood cells within ±500, 400, 300, 200, 100, or 50 or fewer per unit range.

[0200] Alternatively, the level of erythrocyte adhesion to endothelial cells can be determined under flowing conditions. This may involve erythrocyte adhesion to HMECs under flowing conditions, as determined, for example, according to Wautier et al, 2007, Blood, 110:894-901 (the contents of which are incorporated herein by reference). For example, HMECs may be cultured (e.g., in a glass capillary coated with gelatin (2%) for 24 hours), then a microslide containing confluent HMECs is placed on a video microscope stage, and an erythrocyte suspension (e.g., hematocrit 0.5%) is perfused through the microslide at a suitable flow rate (e.g., a rate corresponding to a wall shear stress of 0.03 Pa) for a suitable time (e.g., 10 minutes), followed by washing away unadhered cells (e.g., for 10 minutes). The wall shear stress may then be increased stepwise (e.g., 0.03–0.3 Pa every 5 minutes). In each step, the attached red blood cells are recorded (for example, video imaging with 10 consecutive fields of view, each field of view being 0.1 mm). 2measure it), count it (e.g., using a computerized image analysis system), and express the result as the number of red blood cells per 1 mm 2 It can be expressed as the number of red blood cells per 1 mm

[0201] Under such flow conditions, for example, when analyzing adhesion at 0.03 Pa and washout at 0.07 Pa, in the case of a healthy individual, typically, it can show adhesion of 30 ± 4 red blood cells per 1 mm 2 However, more generally, patients with CRVO can show 190 ± 8 red blood cells per 1 mm 2 Subjects treated according to the present invention, optionally, when evaluated for adhesion to endothelial cells under flow conditions, for example, as determined by techniques such as those described above, may have more than 35 red blood cells per 1 mm 2 such as about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155, about 160, about 165, about 170, about 175, about 180, about 185, about 190, about 195, about 200, or more red blood cells, and can be classified as such. The term "about" in this context refers to a value of ±5, 4, 3, 2, or 1 red blood cell per 1 mm 2 of the stated value

[0202] Under such flow conditions, for example, when analyzing adhesion at 0.03 Pa and washout at 0.3 Pa, in the case of a healthy individual, typically, it can show adhesion of less than 1 red blood cell per 1 mm 2 However, more generally, patients with CRVO can show 20 ± 8 red blood cells per 1 mm 2 Subjects treated according to the present invention, optionally, when evaluated for adhesion to endothelial cells under such flow conditions, for example, as determined by techniques such as those described above, may have 1 mm 2A cell may be classified as having more than one red blood cell per unit, for example, approximately 2, 3, 4, 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 more red blood cells. In this context, the term "approximately" refers to the value in 1 mm. 2 This refers to values ​​where the percentage of red blood cells is ±50%, 40%, 30%, 20%, or 10% or less.

[0203] The retina typically contains three veins associated with retinal vein occlusion: the central retinal vein, the semi-central retinal vein, or the retinal branch veins ("Retinal Vein Occlusion (RVO) Guidelines," The Royal College of Ophthalmologists, July 2015).

[0204] The terms central vein, semi-central vein, and branched central vein provide names for subtypes of retinal vein occlusion, which are central retinal vein occlusion (CRVO), semi-retinal central vein occlusion (HRVO), and branched retinal vein occlusion (BRVO).

[0205] Central retinal vein occlusion is caused by occlusion of the central retinal vein, particularly the portion passing through the lamina cribriformis. Hemicentral retinal vein occlusion is caused by occlusion of the retinal vein in the upper or inner hemisphere of the retina. Branch retinal vein occlusion is caused by occlusion at the arteriovenous crossing where the retinal vein and retinal artery share a common vascular sheath.

[0206] Therefore, in one embodiment, the retinal vein in which RVO exists and / or which is the target of prevention or treatment by the present invention is one or more selected from the central vein, semi-central vein, and branched central vein. In other words, in one embodiment, retinal vein occlusion is one or more of the retinal central vein occlusion, semi-retinal central vein occlusion, and branched retinal vein occlusion.

[0207] Preferably, the retinal vein occlusion is a central retinal vein occlusion.

[0208] Optionally, according to various embodiments of the present invention, treatment of retinal vein occlusion (RVO) or macular edema in a subject having RVO is initiated within 10, 20, 30, 40, 50, or 60 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or within 1, 2, 3, 4, 5, 6, or 7 days of the onset of symptoms of retinal vein occlusion. More preferably, according to the present invention, treatment of retinal vein occlusion or macular edema in subjects with RVO is initiated within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, or 60 minutes, within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or within 1, 2, 3, 4, 5, 6, or 7 days of the diagnosis of retinal vein occlusion. For example, combination therapy may be initiated by the first administration of the annexin A5 component of the co-therapy, or by the first administration of the anti-VEGF component of the co-therapy, or by the simultaneous first administration of both components, either as separate compositions or in combination in the same formulation.

[0209] Retinal vein occlusion can also be classified as either ischemic or non-ischemic. This classification generally relates to areas of non-capillary perfusion, as will be discussed herein. Thus, in one embodiment, retinal vein occlusion is either ischemic (e.g., ischemic central retinal vein occlusion) or non-ischemic (e.g., non-ischemic central retinal vein occlusion).

[0210] Retinal vein occlusion can be further classified as acute retinal vein occlusion. In acute retinal vein occlusion, RVO may develop due to severe ischemia in the retinal veins, and the onset of symptoms is rapid. Therefore, it is necessary to provide treatment as soon as possible after the onset and / or diagnosis of acute retinal vein occlusion. Accordingly, in one embodiment, the retinal vein occlusion is an acute retinal vein occlusion such as acute central retinal vein occlusion. Preferably, treatment of acute retinal vein occlusion according to the present invention is initiated within 10, 20, 30, 40, 50, or 60 minutes, within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or within 1, 2, 3, 4, 5, 6, or 7 days of the onset of symptoms of acute retinal vein occlusion. More preferably, treatment of acute retinal vein occlusion according to the present invention is initiated within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, or 60 minutes of diagnosis of acute retinal vein occlusion, or within 1, 2, 3, 4, 5, or 6 hours. For example, combination therapy may be initiated by the first administration of the annexin A5 component of the cotherapy, or by the first administration of the anti-VEGF component of the cotherapy, or by the simultaneous first administration of both components, either as separate compositions or in combination in the same formulation.

[0211] Retinal vein occlusion can affect either one eye (or one retina), referred to as unilateral retinal vein occlusion, or both eyes (or both retinas), referred to as bilateral retinal vein occlusion. Therefore, in one embodiment, retinal vein occlusion is unilateral retinal vein occlusion (e.g., unilateral central retinal vein occlusion). In another embodiment, retinal vein occlusion is bilateral retinal vein occlusion (e.g., bilateral central retinal vein occlusion).

[0212] Diagnosis and Characterization of Retinal Vein Occlusion Methods for diagnosing retinal vein occlusion, as well as its subtypes, central retinal vein occlusion, semi-central retinal vein occlusion, and branch retinal vein occlusion, are known to those skilled in the art of medicine. Examples of such diagnoses are discussed in the "Retinal Vein Occlusion (RVO) Guidelines" (mentioned above). Generally, patients with retinal vein occlusion will present physicians with visual changes such as blurred vision, loss of part of the visual field, floating spots in the visual field, flashes in the visual field, decreased or reduced vision (i.e., decreased or reduced visual acuity), and loss of vision (i.e., blindness). In some cases, the loss of vision is sudden. In addition, in some cases, the patient does not exhibit any pain.

[0213] Some of the features of retinal vein occlusion that can be used for diagnosis include papilledema (i.e., swelling of the optic nerve head), increased dilation of retinal veins, increased twisting of retinal veins, hemorrhage of the eye, macular edema, bleeding, "cotton balls" (these are white spots on the retina that can be observed on a fundus examination of the retina), retinal edema, electrodiagnostic test (ERG) results, afferent pupillary obstruction, capillary non-perfusion, arteriolar stenosis, venous occlusion, and retinal pigment epithelial changes. These features can generally be observed by a physician by examination of the fundus (the inner surface of the eye opposite the lens) or retina using ophthalmoscopic examination and / or fundus photography. Another diagnostic method used to identify venous occlusion in particular is angiography (such as angiography with a fluorescent dye (e.g., fluorescein)). In fluorescent dye angiography, a dye is injected into the patient, and the behavior of the dye in the veins of the eye can be used to identify whether there is an occlusion in the vein.

[0214] In one embodiment, the subject is diagnosed with retinal vein occlusion, such as central retinal vein occlusion, hemicentral retinal vein occlusion, and bifurcation retinal vein occlusion, preferably central retinal vein occlusion. In an alternative embodiment, the subject is diagnosed with retinal vein occlusion, such as central retinal vein occlusion, hemicentral retinal vein occlusion, and bifurcation retinal vein occlusion, preferably central retinal vein occlusion.

[0215] In one embodiment, the subject has one or more symptoms that are retinal vein occlusion, such as central retinal vein occlusion, hemicentral retinal vein occlusion, and bifurcation retinal vein occlusion, preferably central retinal vein occlusion.

[0216] In one embodiment, retinal vein occlusion is characterized by changes in subjective vision, such as blurred vision, loss of a portion of the visual field, floating spots in the visual field, flashes in the visual field, decreased or diminished vision, and loss of vision. Typically, retinal vein occlusion is characterized by decreased or diminished vision, most typically sudden decreased or diminished vision.

[0217] In another embodiment, retinal vein occlusion is characterized by a significant change in the subject's vision occurring within 7 days, for example, within 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. This may optionally be a characteristic of acute RVO.

[0218] Otherwise, typically, the loss of vision, or the reduction or decline of vision, lasts for about 7 days or more, for example, about 8 days or more, about 9 days or more, about 10 days or more, about 11 days or more, about 12 days or more, about 13 days or more, about 14 days or more, about 15 days or more, about 16 days or more, about 17 days or more, about 18 days or more, about 19 days or more, about 20 days or more, about 21 days or more, about 22 days or more, about 23 days or more, about 24 days or more, about 25 days or more, about 26 days or more, about 27 days or more, about 28 days or more, about 29 days or more, about 30 days or more, about 31 days or more, about 1 month or more, about 2 months or more, about 3 months or more, Occurs over a period of approximately 4 months or more, approximately 5 months or more, approximately 6 months or more, approximately 7 months or more, approximately 8 months or more, approximately 9 months or more, approximately 10 months or more, approximately 11 months or more, approximately 12 months or more, approximately 13 months or more, approximately 14 months or more, approximately 15 months or more, approximately 16 months or more, approximately 17 months or more, approximately 18 months or more, approximately 19 months or more, approximately 20 months or more, approximately 21 months or more, approximately 22 months or more, approximately 23 months or more, approximately 24 months or more, approximately 36 months or more, approximately 3 years or more, approximately 4 years or more, approximately 5 years or more, approximately 6 years or more, approximately 7 years or more, approximately 8 years or more, approximately 9 years or more, or approximately 10 years or more.

[0219] During the period immediately preceding, vision may decline or decrease to the point of complete loss of vision (i.e., blindness) or to a certain percentage of normal vision, at which point that percentage of vision is maintained. Typically, the percentage of vision decline or decrease is approximately 95% or less of normal vision, for example, approximately 90% or less, approximately 85% or less, approximately 80% or less, approximately 75% or less, approximately 70% or less, approximately 69% or less, approximately 68% or less, approximately 67% or less, approximately 66% or less, approximately 65% ​​or less, approximately 64% or less, approximately 63% or less, approximately 62% or less, approximately 61% or less, approximately 60% or less, approximately 59% or less, approximately 58% or less, approximately 57% or less, approximately 56% or less, approximately 55% or less, approximately 54% or less, approximately 53% or less, approximately 52% or less, approximately 51% or less, approximately 50% or less, approximately 49% or less, approximately 48% or less, approximately 47% or less, approximately 46% or less, approximately 45% or less, approximately 44% or less, approximately 43% or less, approximately 42% or less, approximately 41% or less, approximately 40% or less, approximately 39% or less, approximately 38% or less, approximately 37% or less, approximately 36% or less, approximately 35% or less, approximately 34% or less, approximately 33% or less, approximately 32% or less, approximately 31% or less, approximately 30% or less, approximately 25% or less, approximately 20% or less, approximately 15% or less, approximately 10% or less, or approximately 5% or less. Normal visual acuity includes the subject's vision before the onset of retinal vein occlusion or the normal vision of the general population. If the retinal vein occlusion is unilateral retinal vein occlusion, normal visual acuity may be from the eye not affected by the retinal vein occlusion.

[0220] In one embodiment, the subject does not experience any pain in the eye and / or any pain in the retina.

[0221] In one embodiment, retinal vein occlusion is characterized by one or more of the following: papilledema, increased dilation of retinal veins, increased twisting of retinal veins, hemorrhage, "cotton ball" patches, retinal edema, electroreceptor testing (ERG), afferent pupillary obstruction, capillary non-perfusion, arteriolar stenosis, and retinal pigment epithelial changes.

[0222] In one embodiment, retinal vein occlusion (for example, central retinal vein occlusion, preferably ischemic central retinal vein occlusion) is characterized by one or more of the following: • Insufficient visual acuity (for example, less than 95% of normal vision, less than 90% of normal vision, less than 85%, less than 80%, less than 75%, less than 70%, less than 69%, less than 68%, less than 67%, less than 66%, less than 65%, less than 64%, less than 63%, less than 62%, less than 61%, less than 60%, less than 59%, less than 58%, less than 57%, less than 56%, less than 55%, less than 54%, less than 53%, less than 52%, less than 51%) Approximately 50% or less, approximately 49% or less, approximately 48% or less, approximately 47% or less, approximately 46% or less, approximately 45% or less, approximately 44% or less, approximately 43% or less, approximately 42% or less, approximately 41% or less, approximately 40% or less, approximately 39% or less, approximately 38% or less, approximately 37% or less, approximately 36% or less, approximately 35% or less, approximately 34% or less, approximately 33% or less, approximately 32% or less, approximately 31% or less, approximately 30% or less, approximately 25% or less, approximately 20% or less, approximately 15% or less, approximately 10% or less, or approximately 5% or less (preferably approximately 44% or less of normal visual acuity), ·Afferent pupillary disorder, • The presence of one or more retinal hemorrhages, for example, one or more intraretinal hemorrhages (for example, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, nineteen or more, twenty or more retinal hemorrhages), • The presence of one or more "cotton-like white patches" (for example, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, nineteen or more, twenty or more "cotton-like white patches") • Retinal vein dilation, • Twisting of the retinal veins, Angiography showing one or more regions of retinal capillary non-perfusion (for example, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more retinal capillary non-perfusion regions, preferably 10 or more retinal capillary non-perfusion regions), • Electrodiagnostic tests (ERG) show one or more of the following: reduction of b-wave amplitude, reduction of b-wave:a-wave ratio, and extension of b-wave latency, or reduction of b-wave:a-wave ratio and extension of b-wave latency.

[0223] Retinal vein occlusion patient subgroup In addition to the aforementioned patients with different subtypes of retinal vein occlusion (e.g., central retinal vein occlusion), patients with retinal vein occlusion can also be identified by different characteristics.

[0224] Approximately 1% to 2% of red blood cells in patients with retinal vein occlusion are annexin A5 positive (i.e., they have monomeric annexin A5 binding sites such as phosphatidylserine on the outside of the red blood cell membrane), whereas typically, in "normal" patients (e.g., patients without retinal vein occlusion), approximately 0.7% of red blood cells are known to be annexin A5 positive. In comparison, in patients with sickle cell disease, typically approximately 2.84% of red blood cells are annexin A5 positive. Similarly, typically, in patients with retinal vein occlusion, there are an average of approximately 400 to 1000 monomeric annexin A5 binding sites on each annexin A5-positive red blood cell, whereas typically, in "normal" patients, there are an average of approximately 280 monomeric annexin A5 binding sites on each annexin A5-positive red blood cell. In comparison, in patients with sickle cell disease, there are typically an average of approximately 12,340 monomeric annexin A5 binding sites on each annexin A5-positive red blood cell.

[0225] There may also be clear differences among the subjects in the average number of monomeric annexin A5 binding sites exposed on the surface of vascular endothelial cells.

[0226] Microparticles are submicron-sized vesicles that detach from various cell types (e.g., erythrocytes and / or vascular endothelial cells) and are released into the extracellular environment (Hargett and Bauer, 2013, Pulm. Circ., 3(2):329-340). They can typically be detected in the circulating plasma of a subject. The average number of monomeric annexin A5 binding sites exposed on the surface of the microparticles can also vary between subjects, for example, based on the binding site levels in the erythrocytes and / or vascular endothelial cells of the subjects involved in microparticle release.

[0227] Therefore, in one embodiment, the subject is characterized by having about 1% (or 1.0%) to about 2% of cells (e.g., erythrocytes (i.e., red blood cells) and / or vascular endothelial cells) and / or microparticles (e.g., about 1.1% to about 2%, about 1.2% to about 2%, about 1.3% to about 2%, about 1.4% to about 2%, about 1.5% to about 2%, about 1.6% to about 2%, about 1.7% to about 2%, about 1.8% to about 2%, or about 1.9% to about 2% of cells (e.g., red blood cells and / or vascular endothelial cells) and / or microparticles) that are annexin A5 positive. It may be preferable that the subject is characterized by having about 1.7% to about 1.9% of cells (e.g., red blood cells and / or vascular endothelial cells) and / or microparticles that are annexin A5 positive.

[0228] Therefore, the subjects may be characterized by cells (e.g., erythrocytes and / or vascular endothelial cells) and / or microparticles that are annexin A5 positive, comprising approximately 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, or 1.9%, or 2%. It may be preferable that approximately 1.8% of the cells (erythrocytes and / or vascular endothelial cells) and / or microparticles are annexin A5 positive.

[0229] Additionally or alternatively, the subject may be characterized by having cells (e.g., red blood cells and / or vascular endothelial cells) and / or an average number of monomeric annexin A5 binding sites per microparticle of about 400 to about 1000.

[0230] For example, the target is cells (e.g., red blood cells and / or vascular endothelial cells), and / or approximately 410-1000, 420-1000, 430-1000, 440-1000, 450-1000, 460-1000, 470-1000, 480-1000, 490-1000, 500-1000, 510-1000, 520-1000, and 530 particles per microparticle. ~approximately 1000, approximately 540~approximately 1000, approximately 550~approximately 1000, approximately 560~approximately 1000, approximately 570~approximately 1000, approximately 580~approximately 1000, approximately 590~approximately 1000, approximately 600~approximately 1000, approximately 610~approximately 1000, approximately 620~approximately 1000, approximately 630~approximately 1000, approximately 640~approximately 1000, approximately 650~approximately 1000, approximately 660~approximately 1000, approximately 670~approximately 1000, approximately 680~approximately 1000, approximately 690~approximately 1000, approximately 700-1000, approximately 710-1000, approximately 720-1000, approximately 730-1000, approximately 740-1000, approximately 750-1000, approximately 760-1000, approximately 770-1000, approximately 780-1000, approximately 790-1000, approximately 800-1000, approximately 810-1000, approximately 820-1000, approximately 830-1000, approximately 840-1000, approximately 850-10 It may be characterized by having an average number of monomeric annexin A5 binding sites of approximately 0, 860-1000, 870-1000, 890-1000, 900-1000, 910-1000, 920-1000, 930-1000, 940-1000, 950-1000, 960-1000, 970-1000, 980-1000, and 990-1000.

[0231] The target may preferably be cells (e.g., red blood cells and / or vascular endothelial cells) and / or particles having an average number of monomeric annexin A5 binding sites of about 700 to about 800 or about 700 to about 740, most preferably about 710 to about 730 per particle.

[0232] In other words, the target is cells (e.g., red blood cells and / or vascular endothelial cells), and / or approximately 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740 per particle. The particles may be characterized by having an average number of monomeric annexin A5 binding sites of approximately 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, and 990, preferably cells (e.g., red blood cells and / or vascular endothelial cells) and / or approximately 720 monomeric annexin A5 binding sites per microparticle.

[0233] This includes the fact that the binding sites on cells (e.g., erythrocytes and / or vascular endothelial cells) and / or microparticles are located outside the surface membrane (i.e., outer membrane) of the cells or microparticles. Preferably, in the embodiments outlined above, the monomeric annexin A5 binding sites per cell (e.g., erythrocytes and / or vascular endothelial cells) and / or microparticles are located on annexin A5-positive cells and / or microparticles.

[0234] For example, methods for determining the number of annexin A5-positive cells and / or microparticles in a patient by determining the number of annexin A5-positive red blood cells and / or microparticles in the patient's blood are known to those skilled in the art of molecular biology. Various methods are available that allow for the counting of cells and / or microparticles based on markers on the surface of cells and / or microparticles (e.g., flow cytometry). See, for example, Hargett and Bauer, 2013, Pulm. Circ., 3(2):329-340 (the contents of which are incorporated herein by reference). There are also numerous variants of annexin A5 conjugated with fluorescent markers that enable such analysis. A simple example of a possible method is, as known to those skilled in the art, to obtain a sample of the blood of the subject, label it with fluorescent annexin A5, and use a flow cytometer to calculate the number of annexin A5-positive cells and / or microparticles. Thus, "annexin A5-positive" includes cells (such as red blood cells) to which annexin A5 (in particular monomeric annexin A5) can bind.

[0235] In one embodiment, cells (e.g., red blood cells and / or vascular endothelial cells) and / or microparticles are considered annexin A5 positive if they have 290 or more monomeric annexin A5 binding sites, for example, 300 or more, 310 or more, 320 or more, 330 or more, 340 or more, 350 or more, 360 or more, 370 or more, 380 or more, 390 or more, or 400 annexin A5 binding sites.

[0236] However, for the same sample, the average number of monomeric annexin A5 binding sites determined per cell and / or microparticle can vary depending on the specific determination method used. Therefore, in another embodiment, cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles are considered annexin A5 positive if they exceed 100% of the average number of annexin A5 binding sites in the corresponding cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles in a healthy subject. For example, the average number of monomeric annexin A5 binding sites per erythrocyte characteristic of RVO may range from about 1.4 to about 4 times (e.g., about 2 to about 3 times) higher than the average number of monomeric annexin A5 binding sites per erythrocyte in a healthy human.

[0237] One of the molecules to which annexin A5 can bind is phosphatidylserine. The monomeric annexin A5 binding site consists of approximately 30 to 60 phosphatidylserine molecules, and the monomeric annexin A5 binding site is a region of approximately 3,100 angstroms (Å) to 3,300 Å on the surface membrane of cells and / or microparticles. In addition, the monomeric annexin A5 binding site is the region of approximately 40 to 60 phospholipid heads on the surface membrane of cells and / or microparticles.

[0238] In one embodiment, the term "annexin A5 binding site" refers to a region of about 30 to about 60 phosphatidylserine molecules on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles, for example, about 31 to about 59, about 32 to about 58, about 33 to about 57, about 34 to about 56, about 35 to about 55, about 36 to about 54, about 37 to about 53, about 38 to about 52, about 39 to about 51, about 40 to about 50, about 41 to about 49, about 42 to about 48, about 43 to about 47, or about 44 to about 46 phosphatidylserine molecules on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles, preferably about 35 to about 45 phosphatidylserine molecules on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles, more preferably about 42 to about 57 phosphatidylserine molecules on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles, and most preferably about 40 phosphatidylserine molecules on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles.

[0239] In additional or alternative embodiments, the term "annexin A5 binding site" refers to a region of about 3,100 Å to about 3,300 Å on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles, for example, about 3110 Å to about 3290 Å, about 3120 Å to about 3280 Å, about 3130 Å to about 3270 Å, about 3140 Å to about 3260 Å, about 3150 Å to about 3250 Å, about 3160 Å to about 3240 Å, about 3170 Å to about 3230 Å, about 3180 Å to about 3220 Å, about 3190 Å to about 3210 Å on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles, preferably a region of about 3,150 Å to about 3,250 Å on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles, more preferably a region of about 3,000 Å on the surface membrane of cells (e.g., erythrocytes and / or vascular endothelial cells), and / or microparticles.

[0240] In a further embodiment, the term "annexin A5 binding site" refers to about 40 to about 60 phospholipid heads on a region on the surface membrane of a cell (e.g., a red blood cell and / or a vascular endothelial cell) and / or a microparticle, such as about 41 to about 59, about 42 to about 58, about 43 to about 57, about 44 to about 56, about 45 to about 55, about 46 to about 54, about 47 to about 53, about 48 to about 52, about 49 to about 51 phospholipid heads on a region on the surface membrane of a cell (e.g., a red blood cell and / or a vascular endothelial cell) and / or a microparticle, preferably about 45 to about 55 phospholipid heads on a region on the surface membrane of a cell (e.g., a red blood cell and / or a vascular endothelial cell) and / or a microparticle, more preferably about 50 phospholipid heads on a region on the surface membrane of a cell (e.g., a red blood cell and / or a vascular endothelial cell) and / or a microparticle.

[0241] In one embodiment of any of the first, second, third, fourth or fifth aspects of the present invention, the subject is characterized by one or more of the following. a. The subject is of an age of about 50 years or older, such as at least 55 years, 60 years, 65 years, 70 years, 75 years or 80 years (optionally, less than 90 years, such as about 89 years or less, about 88 years or less, about 87 years or less, about 86 years or less, about 85 years or less, about 84 years or less, about 83 years or less, about 82 years or less, about 81 years or less, about 80 years or less, about 79 years or less, about 78 years or less, about 77 years or less, about 76 years or less, about 75 years or less, about 74 years or less, about 73 years or less, about 72 years or less, about 71 years or less, about 70 years or less, about 69 years or less, about 68 years or less, about 67 years or less, about 66 years or less, about 65 years or less, about 64 years or less, about 63 years or less, about 62 years or less, about 61 years or less, about 60 years or less, about 59 years or less, about 58 years or less, about 57 years or less, about 56 years or less, about 55 years or less, about 54 years or less, about 53 years or less, about 52 years or less, about 51 years or less). b. The subjects are those who do not have or are not suspected of having cardiovascular disease (myocardial infarction, stroke, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis), or diseases related to cardiovascular disease (for example, sickle cell anemia, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (Behçet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably diabetes mellitus and diabetes mellitus vera). c. The subjects are those who do not have a history of cardiovascular disease (myocardial infarction, stroke, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis) or cardiovascular disease-related diseases (sickle cell anemia, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (e.g., Behçet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably excluding diabetes mellitus and diabetes mellitus vera), d. The subjects are those who do not have a family history of cardiovascular disease (myocardial infarction, stroke, hypertension, hyperlipidemia, hyperhomocystinemia, blood coagulation disorders, and / or peripheral artery disease, etc., with the exception of atherosclerosis), or a disease related to cardiovascular disease (sickle cell anemia, glaucoma, diabetes mellitus, diabetes mellitus vera, polycythemia vera, and / or systemic inflammatory disorders (e.g., Behcet's disease, polyarteritis nodosa, sarcoidosis, Wegener's granulomatosis, and / or Goodpasture syndrome), but preferably excluding diabetes mellitus and diabetes mellitus vera), and / or e. The subject does not have, or is not suspected of having, one or more conditions selected from solid tumors, lymphomas and / or hematological malignancies, such as breast cancer, endometrial cancer, cervical cancer, ovarian cancer, lung cancer, head and / or cervical cancer, brain cancer, thyroid cancer, esophageal cancer, stomach cancer, colorectal and / or rectal cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, bone cancer, lymphoma (such as Hodgkin lymphoma and / or non-Hodgkin lymphoma), and / or hematological malignancies (such as acute lymphoblastic leukemia, acute myeloid leukemia, and / or chronic myeloid leukemia).

[0242] The "family history" of the subject can be easily determined by a person skilled in the art of medicine. In one embodiment, the family history includes one or more medical histories of the following blood relatives, namely, parents (such as mother or father), grandparents (such as grandmother or grandfather), siblings (such as brother or sister), children (such as son or daughter), and / or grandchildren (such as grandson or granddaughter).

[0243] In one embodiment, if the subject does not exhibit one or more symptoms of cardiovascular disease or a cardiovascular disease-related disorder, the subject is deemed not to have or be suspected of having cardiovascular disease or a cardiovascular disease-related disorder. The symptoms of these disorders are known to those skilled in the art of medicine.

[0244] In one embodiment, the blood coagulation disorder is not retinal vein occlusion such as central retinal vein occlusion, branch retinal vein occlusion, or hemiretinal vein occlusion.

[0245] Preferably, the subject is characterized as follows: a. Be approximately 50 years of age or older. b. The subject does not have or is not suspected of having one or more conditions selected from hypertension, diabetes, and / or sickle cell disease. c. The subject does not have a medical history of one or more conditions selected from hypertension, diabetes, and / or sickle cell disease, and d. The subject does not have a family history of one or more conditions selected from hypertension, diabetes mellitus, and / or sickle cell disease.

[0246] In an alternative embodiment, the subjects may (i) be 50 years of age or older and / or (ii) have a history of suspected hypertension, diabetes mellitus, and / or sickle cell disease, and / or a family history thereof.

[0247] In one embodiment, the subject does not have, or is not suspected of having, a condition characterized by increased extracellular annexin A5. Preferably, the increased extracellular annexin A5 is increased compared to patients who do not have a condition characterized by increased extracellular annexin A5. In one embodiment, the condition characterized by increased extracellular annexin A5 is not retinal vein occlusion and / or a disease associated with retinal vein occlusion.

[0248] It has been demonstrated that a subset of patients with retinal vein occlusion may have a genetic predisposition to form retinal vein occlusion (i.e., possess the genetic risk factors). For example, patients with a genetic predisposition may produce red blood cells that are more prone to thrombus formation in the veins of the eye, particularly in the retina. The formation of retinal vein occlusion in patients with these genetic predispositions is less likely to be associated with other cardiovascular diseases.

[0249] Those in the medical field will know how to identify patients who have a genetic predisposition to develop certain diseases, such as retinal vein occlusion.

[0250] Therefore, in one embodiment, the subject has a genetic predisposition to retinal vein occlusion, or is suspected to have a genetic predisposition to retinal vein occlusion. Preferably, the subject has a genetic predisposition to form one or more thrombi in the veins of the eye (particularly the retina). Most preferably, the subject has a genetic predisposition to produce erythrocytes that are prone to forming one or more thrombi in the veins of the eye (particularly the retina). For example, in vitro bone marrow culture studies in CRVO patients have shown that approximately 27% of CRVO patients exhibit spontaneous proliferation of erythrocyte precursors in the absence of any detectable myeloproliferative disorder (Heron et al, 2007, Ophthalmology, 114:2155-61). Therefore, in one embodiment, the subject may be one that exhibits spontaneous in vitro proliferation of erythrocyte precursors from bone marrow in the absence of any detectable myeloproliferative disorder. Typically, the subject is a human subject.

[0251] In one embodiment, the subject (in particular, a human subject) has a weight of about 10 kg to about 110 kg, for example, about 20 kg to about 100 kg, about 30 kg to about 90 kg, about 40 kg to about 80 kg, or about 50 kg to about 70 kg. In a preferred embodiment, the subject has a weight of about 40 kg to about 80 kg. In another preferred embodiment, the subject has a weight of about 50 kg to about 70 kg. In a more preferred embodiment, the subject has a weight of about 60 kg. When used in this context, the term “about” may be used herein to mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0252] Diseases, disorders, or conditions related to and / or caused by retinal vein occlusion Various diseases, disorders, or conditions are associated with and / or caused by RVO and can be treated by the present invention.

[0253] Thus, in one embodiment, the present invention provides for the prevention or treatment of one or more diseases, disorders, or conditions associated with and / or caused by RVO.

[0254] For example, when a retinal vein is blocked during retinal vein occlusion, blood may not be properly drained and may leak into the retina. When blood leaks into the macula, this can result in macular edema (MO). The leakage worsens depending on the severity of the occlusion, the number of veins involved, and the pressure inside them.

[0255] Macular edema (MO) is the accumulation of fluid in the macula, which is the central area of the retina. The retina is the light-sensitive tissue at the back of the eye, and the macula is the part of the retina responsible for sharp, straight-ahead vision. The accumulation of fluid expands and thickens the macula, distorting vision. Typically, it occurs when there is leakage of fluid and blood into the retina due to an increase in venous pressure and / or capillary pressure that causes vascular permeability. Visual impairment, reduction, or loss in retinal vein occlusion is often associated with macular edema.

[0256] Thus, in one embodiment, the present invention provides a molecule comprising or consisting of monomeric annexin A5 protein for use in the prevention or treatment of macular edema in a subject, wherein the use is preferably in combination therapy with an anti-vascular endothelial growth factor (anti-VEGF) agent in a subject characterized as having, or suspected of having, or at risk of having RVO.

[0257] Other diseases, disorders, or conditions associated with and / or caused by RVO include retinal ischemia, iris neovascularization, retinal neovascularization, neovascular glaucoma, vitreous hemorrhage, rubeosis, and retinal detachment.

[0258] That is, in a separate embodiment, The present invention provides molecules comprising or consisting of annexin A5 protein (such as monomeric annexin A5 protein) for use in the prevention or treatment of retinal ischemia in subjects, preferably as a combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents in subjects characterized by having, suspecting, or being at risk of having RVO.

[0259] The present invention provides molecules containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) for use in the prevention or treatment of diabetic iris neovascularization in subjects, preferably as a combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents in subjects characterized by having, suspecting, or being at risk of having RVO.

[0260] The present invention provides molecules containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) for use in the prevention or treatment of retinal neovascularization in subjects, preferably as a combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents in subjects characterized by having, suspecting, or being at risk of having RVO.

[0261] The present invention provides a molecule containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) for use in the prevention or treatment of neovascular glaucoma in subjects, preferably as a combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents in subjects characterized by having, suspecting, or being at risk of having RVO.

[0262] The present invention provides molecules containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) for use in the prevention or treatment of vitreous hemorrhage in subjects, preferably as a combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents in subjects characterized by having, suspecting, or being at risk of having RVO.

[0263] The present invention provides molecules comprising or consisting of annexin A5 protein (such as monomeric annexin A5 protein) for use in the prevention or treatment of rubeosis in subjects, preferably as a combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents in subjects characterized by having, suspecting, or being at risk of having RVO.

[0264] The present invention provides molecules containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) for use in the prevention or treatment of retinal detachment in subjects, preferably as a combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents in subjects characterized by having, suspecting, or being at risk of having RVO.

[0265] Retinal ischemia is ischemia caused by non-perfusion of retinal veins and capillaries due to retinal vein occlusion.

[0266] Iris neovascularization and retinal neovascularization are the growth of new blood vessels in the eye. These angiogenesis trigger an increase in cytokines (such as VEGF), which act in a positive feedback loop to further promote the growth of new blood vessels.

[0267] Retinal ischemia, iris neovascularization, and one or more of the following can lead to neovascular glaucoma (i.e., damage to the optic nerve), vitreous hemorrhage (i.e., leakage of blood into the vitreous humor of the eye), rubeosis (i.e., formation of new blood vessels in the iris), and / or retinal detachment (i.e., separation of the retina from its blood supply) ("Retinal Vein Occlusion (RVO) Guidelines" (see above)).

[0268] Accordingly, the present invention also provides the prevention or treatment of one or more diseases, disorders, or conditions selected from retinal ischemia, iris neovascularization, retinal neovascularization, neovascular glaucoma, vitreous hemorrhage, rubeosis, and retinal detachment in subjects preferably characterized by having, suspected of having, or at risk of having RVO.

[0269] In one embodiment, iris neovascularization and / or retinal neovascularization is characterized by an increase in cytokines, such as an increase in VEGF. In one embodiment, retinal vein occlusion is central retinal vein occlusion, and the disease associated with retinal vein occlusion is iris neovascularization. In some embodiments, retinal vein occlusion is branch retinal vein occlusion, and the disease associated with retinal vein occlusion is retinal neovascularization. In one embodiment, retinal vein occlusion is hemicentral vein occlusion, and the disease associated with retinal vein occlusion is rubeosis. Preferably, the disease associated with retinal vein occlusion is macular edema.

[0270] In a sixth aspect, the present invention provides a molecule comprising or consisting of annexin A5 protein (such as monomeric annexin A5 protein) for use in the prevention or treatment of diseases, disorders or conditions related to and / or caused by RVO in a subject, wherein this use is in combination with anti-vascular endothelial growth factor (anti-VEGF) agents. Optionally, the composition may contain molecules consisting of monomeric annexin A5 protein in amounts of approximately 50 μg / kg to 100 μg / kg, 40 μg / kg to 80 μg / kg, 30 μg / kg to 60 μg / kg, 20 μg / kg to 40 μg / kg, 1 μg / kg to 30 μg / kg, or less than 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to 10 μg / kg, or approximately 0.01 μg / kg to 1 μg / kg, and / or The composition may optionally contain molecules consisting of monomeric annexin A5 protein in a total amount of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg.

[0271] In an alternative embodiment of the sixth aspect, the present invention provides a method for treating or preventing a disease, disorder or condition related to and / or caused by RVO in a subject, the method comprising administering a therapeutically effective amount of a molecule containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) to the subject. The target group is also treated with an effective prophylactic or therapeutic dose of anti-VEGF agents. Optionally, the composition may contain molecules consisting of monomeric annexin A5 protein in amounts of approximately 50 μg / kg to 100 μg / kg, 40 μg / kg to 80 μg / kg, 30 μg / kg to 60 μg / kg, 20 μg / kg to 40 μg / kg, 1 μg / kg to 30 μg / kg, or less than 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to 10 μg / kg, or approximately 0.01 μg / kg to 1 μg / kg, and / or The composition may optionally contain molecules consisting of monomeric annexin A5 protein in a total amount of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg.

[0272] In an alternative embodiment of the sixth aspect, the present invention provides the use of a molecule comprising or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in the manufacture of a drug for the prevention or treatment of a disease, disorder or condition related to and / or caused by RVO in a subject, This use is in combination with anti-VEGF agents. Optionally, the composition may contain molecules consisting of monomeric annexin A5 protein in amounts of approximately 50 μg / kg to 100 μg / kg, 40 μg / kg to 80 μg / kg, 30 μg / kg to 60 μg / kg, 20 μg / kg to 40 μg / kg, 1 μg / kg to 30 μg / kg, or less than 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to 10 μg / kg, or approximately 0.01 μg / kg to 1 μg / kg, and / or The composition may optionally contain molecules consisting of monomeric annexin A5 protein in a total amount of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg.

[0273] Embodiments of the present invention discussed herein with respect to the first, second, third, and fourth aspects of the present invention are applicable to the sixth aspect of the present invention unless otherwise stated.

[0274] In a seventh aspect, the present invention provides a composition comprising a molecule comprising or consisting of annexin A5 protein (such as monomeric annexin A5 protein), Optionally, the composition may contain molecules consisting of monomeric annexin A5 protein in amounts of approximately 50 μg / kg to 100 μg / kg, 40 μg / kg to 80 μg / kg, 30 μg / kg to 60 μg / kg, 20 μg / kg to 40 μg / kg, 1 μg / kg to 30 μg / kg, or less than 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to 10 μg / kg, or approximately 0.01 μg / kg to 1 μg / kg, and / or Optionally, the composition may contain molecules consisting of monomeric annexin A5 protein in a total amount of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg, and Optionally, the composition is intended for use in the prevention or treatment of one or more diseases, disorders, or conditions related to and / or caused by RVO in the subject, and this use is intended as combination therapy with anti-vascular endothelial growth factor (anti-VEGF) agents.

[0275] In an alternative embodiment of the seventh aspect, the present invention provides a method for treating or preventing one or more diseases, disorders or conditions related to and / or caused by RVO in a subject, the method comprising administering to a subject a therapeutically effective amount of a composition comprising a molecule comprising or consisting thereof annexin A5 protein (such as monomeric annexin A5 protein), The target group is also treated with an effective prophylactic or therapeutic dose of anti-VEGF agents. Optionally, the composition may contain molecules consisting of monomeric annexin A5 protein in amounts of approximately 50 μg / kg to 100 μg / kg, 40 μg / kg to 80 μg / kg, 30 μg / kg to 60 μg / kg, 20 μg / kg to 40 μg / kg, 1 μg / kg to 30 μg / kg, or less than 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to 10 μg / kg, or approximately 0.01 μg / kg to 1 μg / kg, and / or The composition may optionally contain molecules consisting of monomeric annexin A5 protein in a total amount of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg.

[0276] Embodiments of the first, second, third, fourth, and sixth aspects of the present invention discussed herein are applicable to the seventh aspect of the present invention unless otherwise stated.

[0277] C. Annexin A5 Molecules containing annexin A5 protein, such as monomeric annexin A5 protein, for use in all embodiments of the present invention may include proteins consisting of an annexin A5 sequence, such as a human annexin A5 sequence.

[0278] A molecule comprising monomeric annexin A5 protein for use in all aspects of the present invention may include, essentially consist of, or be composed of a protein comprising an annexin A5 sequence, such as a human annexin A5 sequence.

[0279] To avoid misunderstanding, the molecule consisting of monomeric annexin A5 protein is not dimeric annexin A5 (commonly referred to as "dianexin" in the art) nor is it PEGylated annexin A5.

[0280] Preferably, a molecule containing or consisting of human annexin A5 protein (such as human monomeric annexin A5 protein) is a protein that contains, consists of, or is essentially composed of a polypeptide molecule having the amino acid sequence of SEQ ID NO: 1, either containing or not containing an N-terminal methionine, as shown below. Preferably, a molecule containing, consisting of, or essentially composed of annexin A5 protein (such as monomeric annexin A5 protein) is a recombinant human annexin A5 protein (such as recombinant human monomeric annexin A5 protein), for example, a protein produced by recombinantly expressing a gene encoding a protein having the sequence of SEQ ID NO: 1 in a recombinant host cell organism such as an Escherichia coli (E. coli) host, and preferably subsequently recovering and purifying the annexin A5 protein therefrom.

[0281] The sequence of the protein encoded by the human annexin A5 gene is defined by sequence number 1, as shown below.

[0282] [Table 1]

[0283] In another embodiment, the annexin A5 (monomer annexin A5, for example) protein may include, be essentially, or be a variant of the annexin A5 sequence, such as the sequence of human annexin A5 (optionally, a protein consisting of the sequence of SEQ ID NO: 1, with or without an N-terminal methionine). For example, the sequence of a variant or mutant may differ from the sequence of SEQ ID NO: 1 (with or without the N-terminal methionine) at one or more positions, for example, at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160 or earlier, or at positions up to or before those positions.

[0284] In another embodiment, the annexin A5 (such as monomeric annexin A5) protein may include, be essentially, or be derived from, a biologically active fragment of mature annexin A5, such as human annexin A5 (optionally, a protein consisting of the sequence of SEQ ID NO: 1, with or without an N-terminal methionine), or a variant or mutant thereof.

[0285] Biologically active fragments of annexin A5 share functional or binding properties with full-length annexin A5. Epitope fragments of annexin A5 bind to monoclonal antibodies that bind to full-length annexin A5. "Activity" of annexin A5 refers to any binding function performed by the protein, further examples of which are defined below.

[0286] The biologically active fragments of monomeric annexin A5 protein that can be used in the present invention may include, but not limited to, human annexin A5 protein (optionally, a protein consisting of the sequence of SEQ ID NO: 1, with or without an N-terminal methionine) or a modification thereof, at least about 50 consecutive amino acids, usually at least about 100 consecutive amino acids, at least about 150 consecutive amino acids, at least about 200 consecutive amino acids, at least about 250 consecutive amino acids, at least about 260 consecutive amino acids, at least about 270 consecutive amino acids, at least about 280 consecutive amino acids, at least about 290 consecutive amino acids, at least about 300 consecutive amino acids, at least about 310 consecutive amino acids, and may include up to about 320 consecutive amino acids, and may further include fusion polypeptides known in the art in addition to the provided sequence. The annexin A5 sequence can originate from any mammalian or bird species, such as primates, particularly rodents including humans, mice, rats, and hamsters, rabbits, horses, cattle, dogs, and cats. Of particular interest is the human annexin A5 protein (a protein consisting of the sequence of Sequence ID No. 1, with or without N-terminal methionine, as optional).

[0287] In one embodiment, a biologically active fragment of annexin A5 (such as monomeric annexin A5) protein is effective in treating retinal vein occlusion or diseases associated with retinal vein occlusion, and is preferably 10% or more effective compared to human monomeric annexin A5 protein, which is a polypeptide molecule consisting of the amino acid sequence of SEQ ID NO: 1, with or without an N-terminal methionine, for example, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more effective.

[0288] In one embodiment, the monomeric annexin A5 protein consists of approximately 320 or fewer amino acids, for example, approximately 319 or fewer amino acids, approximately 318 or fewer amino acids, approximately 317 or fewer amino acids, approximately 316 or fewer amino acids, approximately 315 or fewer amino acids, approximately 314 or fewer amino acids, approximately 313 or fewer amino acids, approximately 312 or fewer amino acids, approximately 311 or fewer amino acids, approximately 310 or fewer amino acids, approximately 305 or fewer amino acids, approximately 300 or fewer amino acids, approximately 295 or fewer amino acids, approximately 290 or fewer amino acids, approximately 285 or fewer amino acids, approximately 280 or fewer amino acids, approximately 270 or fewer amino acids, approximately 260 or fewer amino acids, or approximately 250 or fewer amino acids.

[0289] In one embodiment, the recombinant monomer annexin A5 protein consists of 319 amino acids, for example, about 318 or fewer amino acids, about 317 or fewer amino acids, about 316 or fewer amino acids, about 315 or fewer amino acids, about 314 or fewer amino acids, about 313 or fewer amino acids, about 312 or fewer amino acids, about 311 or fewer amino acids, about 310 or fewer amino acids, about 305 or fewer amino acids, about 300 or fewer amino acids, about 295 or fewer amino acids, about 290 or fewer amino acids, about 285 or fewer amino acids, about 280 or fewer amino acids, about 270 or fewer amino acids, about 260 or fewer amino acids, or about 250 or fewer amino acids.

[0290] Annexin A5 (monomer annexin A5, etc.) proteins may contain one or more of the following: a. A protein that contains, is essentially derived from, or consists of the sequence of human monomeric annexin A5 protein (optionally, a protein consisting of the sequence of SEQ ID NO: 1, with or without an N-terminal methionine). b. Recombinant human annexin A5 proteins such as human monomer annexin A5 protein, c. Mammalian orthologs of human monomer annexin A5 protein, Alleles or genetic variants of da), b), or c), Functional analogues or variants of annexin A5 proteins, such as monomeric annexin A5 protein, which is a protein having the sequence defined by SEQ ID NO: 1, with or without an eN-terminal methionine, and comprising 50%, 60%, 70%, 75%, for example, more than 80%, 85%, 90%, or more preferably more than 95%, or 99% of the amino acid sequence identity of human monomeric annexin A5 protein, and One of the biologically active fragments from fa to e).

[0291] Molecules comprising annexin A5 protein may also be referred to herein as “agents containing annexin A5 protein” or “therapeutic agents containing annexin A5 protein,” and may also be referred to as “annexin A5.” Optionally, the term “molecule containing annexin A5 protein” may exclude molecules containing forms of annexin A5 other than monomeric annexin A5, such as dimeric monomeric annexin A5 protein (also known as dianexin).

[0292] In some embodiments, the molecule containing annexin A5 protein for use in the present invention may be a conjugate in which annexin A5 protein (such as monomeric annexin A5 protein) is conjugated to a portion such as a non-proteinoid portion. Non-limitingly, this portion may be, for example, a therapeutic, prophylactic, diagnostic, prognostic, or theragnotic portion. In some embodiments, this portion may be a drug (such as a non-proteinoid drug) and / or a radioactive portion (for example, if the conjugate is suitable for use in radioimmunotherapy ("RIT").

[0293] Molecules comprising monomeric annexin A5 protein may also be referred to herein as “agents comprising monomeric annexin A5 protein” or “therapeutic agents comprising monomeric annexin A5 protein” and may also be referred to as “monomeric annexin A5.” The purpose of the term “molecule comprising monomeric annexin A5 protein” is to exclude molecules that are not monomeric annexin A5, such as PEGylated annexin A5, dimeric monomeric annexin A5 protein (also known as dianexin), and annexin A5 fusion proteins, in particular annexin A5 fusion proteins that have a higher weight-average molecular weight and / or plasma half-life compared to human monomeric annexin A5 protein, which is a polypeptide molecule comprising the amino acid sequence of SEQ ID NO: 1, with or without an N-terminal methionine.

[0294] Therefore, a molecule consisting of monomeric annexin A5 protein is not PEGylated annexin A5, dimeric monomeric annexin A5 protein, or annexin A5 fusion protein. To avoid misunderstanding, in one embodiment, a molecule consisting of monomeric annexin A5 protein is not a “modified annexin” as defined in U.S. Patent No. 9,463,217 (the contents of which are incorporated herein by reference in their entirety).

[0295] In certain embodiments, functional analogs, variants, or derivatives of annexin A5 according to the present invention are either containing or not containing an N-terminal methionine, and are 50%, 60%, 70%, 75%, for example, 80%, 85%, 90%, or more preferably 95%, or 99% identical to human annexin A5, SEQ ID NO: 1.

[0296] The percentage of identity between two amino acid sequences is determined as follows: First, the amino acid sequences are compared, for example, SEQ ID NO: 1, using the BLAST 2 Sequences (Bl2seq) program from the standalone version of BLASTZ, which includes BLASTN version 2.0.14 and BLASTP version 2.0.14. This standalone version of BLASTZ is available from the website of the U.S. National Center for Biotechnology Information (ncbi.nlm.nih.gov). Instructions on how to use the Bl2seq program can be found in the readme file included with BLASTZ. Bl2Seq uses the BLASTP algorithm to compare the two amino acid sequences. To compare the two amino acid sequences, the Bl2seq options are set as follows: -i is set to the file containing the first amino acid sequence to be compared (e.g., C:\seq1.txt), -j is set to the file containing the second amino acid sequence to be compared (e.g., C:\seq2.txt), -p is set to blastp, -o is set to any desired filename (e.g., C:\output.txt), and all other options remain at their default settings. For example, the following command can be used to generate an output file containing a comparison between two amino acid sequences: C:\Bl2seq -ic:\seq1.txt -jc:\seq2.txt -p blastp -oc:\output.txt. If the two compared sequences share homology, the specified output file will present those homologous regions as aligned sequences. If the two compared sequences do not share homology, the specified output file will not present aligned sequences. Once aligned, the number of matches is determined by counting the number of positions where identical nucleotides or amino acid residues exist in both sequences.

[0297] The identity percentage is calculated by dividing the number of matches by the length of the sequence shown in the identified sequence, and then multiplying the resulting value by 100. For example, if a sequence is compared to the sequence shown in sequence number 1 (the length of the sequence shown in sequence number 1 is 320), and the number of matches is 288, then the sequence has an identity percentage of 90 (i.e., 288 ÷ 320 × 100 = 90) with respect to the sequence shown in sequence number 1.

[0298] Therefore, a functional analogue, mutant, or variant of annexin A5 may be a protein having one or more amino acid insertions, deletions, or substitutions, either conserved or nonconserved, provided that such changes result in a protein whose fundamental properties for functioning equivalently to annexin A5 are not significantly altered. In this context, "significantly" means that the properties of the variant may still be different, but not obvious to those of the original protein, as would be said by a person skilled in the art.

[0299] "Conservative substitutions" refer to combinations such as Gly, Ala; Val, Ile, Leu; Asp, Glu; Asn, Gln; Ser, Thr; Lys, Arg; and Phe, Tyr.

[0300] Such variants and mutants can be produced using protein engineering and site-directed mutagenesis methods known in the art.

[0301] Functional analogues, variants, or biologically active fragments of annexin A5 may optionally consist of sequences of human annexin A5 (e.g., SEQ ID NO: 1) that contain or do not contain an N-terminal methionine, having 50, 40, 30, 20, 10, 5, 4, 3, 2, or one or fewer consecutive or discontinuous additional amino acids, and / or 50, 40, 30, 20, 10, 5, 4, 3, 2, or one or fewer consecutive or discontinuous amino acid deletions, and / or 50, 40, 30, 20, 10, 5, 4, 3, 2, or one or fewer consecutive or discontinuous amino acid substitutions.

[0302] D. Target of treatment Subjects treated according to the present invention may be human or non-human subjects. Preferably, subjects according to any aspect of the present invention are characterized by having, being suspected of having, or at risk of having, a condition related to macular edema and / or RVO, or either or both of macular edema and / or RVO.

[0303] The subjects may, at their discretion and not necessarily, be treatment-inexperienced in the sense that they have not previously taken any form of therapeutic agent for the specific condition being treated by the present invention.

[0304] In a preferred embodiment, the subject is a human subject.

[0305] In another embodiment, the subject is a non-human subject and may be selected from a group consisting of non-human mammals, birds, amphibians, and reptiles.

[0306] Non-human mammals may be selected from a list consisting of equids (e.g., horses or donkeys), bovines (e.g., cows, buffaloes, bison, or yaks), camels, pigs, llamas, alpacas, birds (e.g., ostriches, chickens, geese, ducks, turtles, quail, or pigeons), sheep, goats, dogs, cats, reptiles (e.g., snakes, lizards, crocodiles, alligators, turtles, or sea turtles), rabbits, amphibians (e.g., frogs or salamanders), or rodents (e.g., mice, rats, chinchillas, guinea pigs, or squirrels).

[0307] E. Dosage regimen Dosage and timing: According to all aspects of the present invention, with respect to any given administration round, a molecule comprising or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered to the target. The dosage may be optionally determined based on the subject's body weight, and / or be administered in doses of approximately 50 μg / kg to 100 μg / kg, 40 μg / kg to 80 μg / kg, 30 μg / kg to 60 μg / kg, 20 μg / kg to 40 μg / kg, 1 μg / kg to 30 μg / kg, or less than 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to 10 μg / kg, or approximately 0.01 μg / kg to 1 μg / kg of molecular weight, and / or The total dose may be administered at the discretion of the patient, regardless of their body weight, as defined above, at approximately 5 mg, 4 mg, 3 mg, 2 mg, or less than 2 mg, for example, approximately 1 mg, 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg.

[0308] (a) Dosage per body weight: To avoid misunderstanding, please understand that the term "mg / kg body weight" refers to the number of milligrams of molecules containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) per kilogram of body weight of the subject.

[0309] Preferred doses may be selected from one or more of the following ranges: approximately 50 μg / kg to approximately 100 μg / kg, approximately 40 μg / kg to approximately 80 μg / kg, approximately 30 μg / kg to approximately 60 μg / kg, approximately 20 μg / kg to approximately 40 μg / kg, approximately 1 μg / kg to approximately 30 μg / kg, less than approximately 20 μg / kg, for example, approximately 0.01 μg / kg to approximately 10 μg / kg, or approximately 0.01 μg / kg to approximately 1 μg / kg, for example, approximately 0.01 μg / kg, approximately 0.02 μg / kg, approximately 0.03 μg / kg, approximately 0.04 μg / kg, approximately 0.05 μg / kg, approximately 0.06 μg / kg, approximately 0.07 μg / kg, approximately 0.08 μg / kg, approximately 0.09 μg / kg, and / or approximately 0.1 μg / kg.

[0310] In some embodiments, the dose may be selected from one or more of approximately 0.1 μg / kg, approximately 0.2 μg / kg, approximately 0.3 μg / kg, approximately 0.4 μg / kg, approximately 0.5 μg / kg, approximately 0.6 μg / kg, approximately 0.7 μg / kg, approximately 0.8 μg / kg, approximately 0.9 μg / kg, and / or approximately 1 μg / kg.

[0311] In other embodiments, the dose may be selected from one or more of approximately 1 μg / kg, approximately 2 μg / kg, approximately 3 μg / kg, approximately 4 μg / kg, approximately 5 μg / kg, approximately 6 μg / kg, approximately 7 μg / kg, approximately 8 μg / kg, approximately 9 μg / kg, and / or approximately 10 μg / kg.

[0312] Pharmaceutically or veterinarily acceptable compositions containing or comprising molecules of annexin A5 protein (such as monomeric annexin A5 protein) may also be administered as defined above, and the composition may be administered such that the amount of molecules containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) in the composition is about 5 mg, about 4 mg, about 3 mg, about 2 mg, less than about 2 mg, for example, about 1 mg, about 0.1 to 1.5 mg, 0.001 mg to about 1 mg, or about 0.001 mg to about 0.1 mg.

[0313] In one embodiment, the subject is administered a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in an amount ranging from approximately 0.00001 mg / kg body weight to approximately "X" mg / kg body weight, where "X" is approximately 1 mg / kg, 0.5 mg / kg, 0.4 mg / kg, 0.3 mg / kg, 0.2 mg / kg, 0.1 mg / kg, 0.09 mg / kg, 0.08 mg / kg, 0.07 mg / kg, 0.06 mg / kg. g / kg, 0.05 mg / kg, 0.04 mg / kg, 0.03 mg / kg, 0.02 mg / kg, 0.01 mg / kg, 0.009 mg / kg body weight or less, for example: approximately 0.008 mg / kg body weight or less, approximately 0.007 mg / kg body weight or less, approximately 0.006 mg / kg body weight or less, approximately 0.005 mg / kg body weight or less, approximately 0.004 mg / kg body weight or less, approximately 0.003 mg / kg body weight or less, approximately 0.002 mg / kg body weight or less, Preferably, about 0.001 mg / kg body weight or less, 0.0009 mg / kg body weight or less, for example, about 0.0008 mg / kg body weight or less, about 0.0007 mg / kg body weight or less, about 0.0006 mg / kg body weight or less, about 0.0005 mg / kg body weight or less, about 0.0004 mg / kg body weight or less, about 0.0003 mg / kg body weight or less, about 0.0002 mg / kg body weight or less, about 0.0001 mg / kg body weight or less, about 0.00009 A molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in amounts of less than or equal to mg / kg body weight, approximately less than or equal to 0.00008 mg / kg body weight, approximately less than or equal to 0.00007 mg / kg body weight, approximately less than or equal to 0.00006 mg / kg body weight, approximately less than or equal to 0.00005 mg / kg body weight, approximately less than or equal to 0.00004 mg / kg body weight, approximately less than or equal to 0.00003 mg / kg body weight, or approximately less than or equal to 0.00002 mg / kg body weight.

[0314] In additional or alternative embodiments, the subject may contain or consist of molecules containing annexin A5 protein (such as monomeric annexin A5 protein) in amounts of approximately "Y" mg / kg body weight to approximately 0.01 mg / kg body weight, preferably approximately "Y" mg / kg body weight to approximately 1 mg / kg, 0.5 mg / kg, 0.4 mg / kg, 0.3 mg / kg, 0.2 mg / kg, 0.1 mg / kg, 0.09 mg / kg, 0.08 mg / kg, or 0.07 mg / kg. , administered in amounts of 0.06 mg / kg, 0.05 mg / kg, 0.04 mg / kg, 0.03 mg / kg, 0.02 mg / kg, or 0.001 mg / kg body weight, and "Y" is approximately 0.00002 mg / kg body weight or more, for example: approximately 0.00003 mg / kg body weight or more, approximately 0.00004 mg / kg body weight or more, approximately 0.00005 mg / kg body weight or more, approximately 0.00006 mg / kg body weight or more, approximately 0.00007 mg / kg body weight or more, approximately 0. 0.0008 mg / kg body weight or more, approximately 0.00009 mg / kg body weight or more, approximately 0.0001 mg / kg body weight or more, approximately 0.0002 mg / kg body weight or more, approximately 0.0003 mg / kg body weight or more, approximately 0.0004 mg / kg body weight or more, approximately 0.0005 mg / kg body weight or more, approximately 0.0006 mg / kg body weight or more, approximately 0.0007 mg / kg body weight or more, approximately 0.0008 mg / kg body weight or more, or approximately 0.0009 mg / kg body weight or more, approximately 0.0 A molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in amounts of 0.01 mg / kg body weight or more, approximately 0.002 mg / kg body weight or more, approximately 0.003 mg / kg body weight or more, approximately 0.004 mg / kg body weight or more, approximately 0.005 mg / kg body weight or more, approximately 0.006 mg / kg body weight or more, approximately 0.007 mg / kg body weight or more, approximately 0.008 mg / kg body weight or more, or approximately 0.009 mg / kg body weight or more.

[0315] In additional or alternative embodiments, subjects are administered a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in an amount of approximately "X" mg / kg body weight to approximately "Y" mg / kg body weight, where "X" and "Y" are the values ​​defined above, respectively.

[0316] The aforementioned dosage per body weight may be particularly suitable for human subjects.

[0317] (b) Total dose: Additionally or alternatively, the dosage may be determined independently of the subject's body weight.

[0318] The preferred dose may be a total dose selected from one or more of the following: approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg. The dose is optional and may be: Approximately 1 μg, approximately 2 μg, approximately 3 μg, approximately 4 μg, approximately 5 μg, approximately 6 μg, approximately 7 μg, approximately 8 μg, approximately 9 μg and / or approximately 10 μg, Approximately 10 μg, approximately 20 μg, approximately 30 μg, approximately 40 μg, approximately 50 μg, approximately 60 μg, approximately 70 μg, approximately 80 μg, approximately 90 μg, and / or approximately 100 μg, It may be approximately 100 μg, approximately 200 μg, approximately 300 μg, approximately 400 μg, approximately 500 μg, approximately 600 μg, approximately 700 μg, approximately 800 μg, approximately 900 μg, and / or approximately 1 mg.

[0319] The aforementioned total dose may be particularly suitable for human subjects, especially adult human subjects, and / or human subjects having a body weight of about 30 kg to about 200 kg, for example, about 40 kg to about 150 kg, or 50 kg to about 120 kg, for example, about 60 kg to about 100 kg.

[0320] In additional or alternative embodiments, the subject is administered a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in an amount of about 0.001 mg to about "A" mg, where "A" is about 5 mg, about 4 mg, about 3 mg, about 2 mg, less than about 2 mg, for example about 1 mg, about 0.9 mg or less, for example about 0.8 mg or less, about 0.7 mg or less, about 0.6 mg or less, about 0.5 mg or less, about 0.4 mg or less, about 0.3 mg or less, about 0.2 mg or less, more preferably about 0.1 mg or less, about 0.09 mg A molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in amounts of g or less, for example, approximately 0.08 mg or less, approximately 0.07 mg or less, approximately 0.06 mg or less, approximately 0.05 mg or less, approximately 0.04 mg or less, approximately 0.03 mg or less, approximately 0.02 mg or less, approximately 0.01 mg or less, approximately 0.009 mg or less, approximately 0.008 mg or less, approximately 0.007 mg or less, approximately 0.006 mg or less, approximately 0.005 mg or less, approximately 0.004 mg or less, approximately 0.003 mg or less, or approximately 0.002 mg or less.

[0321] In additional or alternative embodiments, the subject is administered a molecule containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) in an amount of approximately "B" mg to approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, less than approximately 2 mg, for example, approximately 1 mg or approximately 0.1 mg, where "B" is approximately 0.002 mg or more, for example: approximately 0.003 mg or more, approximately 0.004 mg or more, approximately 0.005 mg or more, approximately 0.0 A molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in an amount of 0.6 mg or more, approximately 0.007 mg or more, approximately 0.008 mg or more, approximately 0.009 mg or more, approximately 0.01 mg or more, approximately 0.02 mg or more, approximately 0.03 mg or more, approximately 0.04 mg or more, approximately 0.05 mg or more, approximately 0.06 mg or more, approximately 0.07 mg or more, approximately 0.08 mg or more, or approximately 0.09 mg or more.

[0322] In additional or alternative embodiments, the subject is administered a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in an amount of approximately "B" mg to approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, less than approximately 2 mg, for example, approximately 1 mg, where "B" is approximately 0.002 mg or more, for example, approximately 0.003 mg or more, approximately 0.004 mg or more, approximately 0.005 mg or more, approximately 0.006 mg or more, approximately 0.007 mg or more, approximately 0.008 mg or more, approximately 0.009 mg or more, approximately 0. A molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in amounts of 0.01 mg or more, approximately 0.02 mg or more, approximately 0.03 mg or more, approximately 0.04 mg or more, approximately 0.05 mg or more, approximately 0.06 mg or more, approximately 0.07 mg or more, approximately 0.08 mg or more, approximately 0.09 mg or more, approximately 0.1 mg or more, approximately 0.2 mg or more, approximately 0.3 mg or more, approximately 0.4 mg or more, approximately 0.5 mg or more, approximately 0.6 mg or more, approximately 0.7 mg or more, approximately 0.8 mg or more, or approximately 0.9 mg or more.

[0323] In additional or alternative embodiments, the subject is administered a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) in an amount of approximately "A" mg to approximately "B" mg, where "A" and "B" are the values ​​defined above, respectively.

[0324] With respect to all other aspects of the present invention, the embodiments of the invention discussed herein are applicable to the fifth, sixth and / or seventh aspects of the invention unless otherwise stated.

[0325] Route of administration: In one preferred embodiment in the context of the prevention or treatment of macular edema, RVO, macular edema or RVO-related conditions, or any other ocular condition, a molecule or composition containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) is administered distally from the eye. Distal administration has the advantage of avoiding the administration of molecules containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) into the eye of the subject, which may be unpleasant for the subject. In particular, there are numerous potential complications associated with intravitreal injection (e.g., using anti-VEGF therapy established for the treatment of macular edema and / or RVO), including infectious endophthalmitis, aseptic endophthalmitis, and retinal vasculitis, particularly brolucizumab (Cox et al., 2021 (see above)), and transient intraocular pressure (IOP) spikes (Levin et al., 2021, J Glaucoma, 30:1019-26).

[0326] In one preferred embodiment in the context of preventing or treating macular edema, RVO, conditions associated with macular edema or RVO, or any other ocular condition, one or more of the administrations of anti-VEGF agents to the subject may be performed directly into the eye, for example, by intravitreal injection. Given the numerous potential complications associated with intravitreal injection of anti-VEGF agents, the objective of the co-therapeutic approach of the present invention is to enhance the preventive and / or therapeutic benefits of anti-VEGF therapy using molecules containing or comprising annexin A5 protein (such as monomeric annexin A5 protein), and / or to mitigate complications associated with the requirements for administering anti-VEGF agents by, for example, reducing the number of rounds of anti-VEGF administration, reducing the frequency of anti-VEGF administration, and / or shortening the period during which anti-VEGF agents must be administered to the subject, as required to achieve the desired preventive and / or therapeutic benefits or preventive and / or therapeutic endpoints.

[0327] According to one preferred option of all embodiments of the present invention, a molecule or composition containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) is administered systemically or by choroidal injection. Preferably, the molecule or composition containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) is administered to the target vascular system. Most preferably, the molecule or composition containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) is administered to the target blood.

[0328] In one embodiment, a molecule containing or comprising annexin A5 protein (such as monomeric annexin A5 protein), or a composition thereof, is administered by injection, blood transfusion, or infusion, for example, intravenously or intra-arterially (particularly intravenous injection or intra-arterial injection).

[0329] In preferred embodiments, molecules or compositions containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) are administered by injection.

[0330] In an alternative, preferred embodiment, a molecule or composition containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) is administered by intravenous injection.

[0331] In a more preferred embodiment, a molecule or composition containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) is administered into the target blood by injection, for example, intravenous injection.

[0332] In another embodiment, molecules or compositions containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) are administered parenterally, intrathecally, intraperitoneally, intraarticularly, intramuscularly, subcutaneously, or topically (however, as described above, in one embodiment, direct administration to the eye can be optionally avoided).

[0333] Alternatively, though less preferred, a molecule containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) may be administered directly to the eye. For example, in one optional and preferred embodiment, a molecule containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) may be administered to one or both of the target eyes by intravitreous injection. According to this embodiment, annexin A5 protein may be administered to the patient, for example, in the form of a sterile intravitreous composition for injection as defined in a fourth aspect of the present invention.

[0334] The composition for intravitreal injection may be present, for example, in a sterile syringe in a form ready for immediate injection (e.g., without requiring reconstitution with a carrier or diluent). Preferably, the syringe contains the composition in an amount of about 200 μL or less, preferably about 100 μL or less, and more preferably the amount is about 10 to about 90 μL, about 20 to about 80 μL, about 30 to about 70 μL, about 40 to about 60 μL, or about 50 μL (in this context, the term "about" refers to an amount of ±4, 3, 2, or 1 μL of the stated value).

[0335] Compositions for intravitreal injection may be provided, for example, in the form of a kit containing the composition and one or more additional components.

[0336] For example, a kit may include a composition present in a sterile syringe and one or more sterile needles attached thereto or separately contained within the kit. Similarly, various embodiments of the present invention may include intravitreal injection of a composition comprising a molecule consisting of an effective amount of monomeric annexin A5 protein into one or both of the target eyes via a sterile needle.

[0337] Preferably, the needle or each needle may be a gauge selected from gauges 27, 28, 29, or more preferably 30 or 31. The needle or each needle may preferably have a length of about 0.5 to 0.62 inches (12.7 to 15.75 mm). Longer needles may increase the risk of retinal damage if the patient accidentally moves forward during the intravitreal injection procedure.

[0338] In less preferred embodiments in which the composition is administered to a subject by intravitreal injection, the procedure may include one or more pre-injection risk control steps selected from: 1) applying a local anesthetic; 2) applying an agent suitable for reducing the risk of bacterial colonization (povidone-iodine eye drops and / or periorbital povidone-iodine eyelid preparations may be particularly preferred); 3) inserting a sterile eyelid retractor to separate the eyelids; and 4) optionally reapplying an agent suitable for reducing the risk of bacterial colonization (e.g., povidone-iodine) just above the injection site before injection.

[0339] In embodiments where intravitreal injection is performed together with local anesthesia, local anesthetic eye drops can be conveniently used.

[0340] Any suitable agent for reducing the risk of bacterial colonization may be used in the eye as part of a pre-injection risk management protocol. Povidone-iodine is one preferred option. Povidone-iodine has been shown to reduce the risk of bacterial colonization and endophthalmitis. It is recommended to apply povidone-iodine to the conjunctival surface, eyelids, and eyelashes before intravitreal injection, preferably also before introducing a sterile eyelid retractor. (The eyelid retractor prevents the needle tip from touching the eyelid or eyelashes before needle insertion.) Povidone-iodine may be used in 5% to 10% solutions, optionally. Studies have found that a 5% povidone-iodine solution is as effective as a 10% solution and causes less irritation to the eye. Optionally, any suitable agent for reducing the risk of bacterial colonization (e.g., povidone-iodine) may be reapplied immediately above the injection site before injection.

[0341] Antibiotics may be used before, during, and / or after intravitreal injection. Any suitable antibiotic may be used. One exemplary embodiment is a fourth-generation fluoroquinolone.

[0342] In another example, the kit may comprise one or more further compositions, each containing one or more antibiotics, for intravitreal injection and / or subsequent administration to a patient.

[0343] Those skilled in the art can select a suitable intravitreal injection site. For example, in one exemplary embodiment, the patient is instructed to avert their gaze from the needle entry site, and the injection is placed approximately 3 to 3.5 mm posterior to the limbus (e.g., in the case of aphakia or pseudophakia) or approximately 3.5 to 4 mm posterior to the limbus (e.g., in the case of phakic eye). Injections in the inferior temporal quadrant are common, but any quadrant (e.g., the superior temporal quadrant) may be used.

[0344] Alternatively, a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5) may be directly administered to one or both retinal veins of the target eye (such as the central retinal vein, retinal branch veins, and / or hemiretinal veins), for example, by catheterization.

[0345] Dosage schedule: In light of the disclosure of the present invention and by applying the general knowledge common thereto, a person skilled in the art can easily determine, at their discretion, a suitable dose for achieving the desired therapeutic effect within the molecular range of approximately 50 μg / kg to approximately 100 μg / kg, approximately 40 μg / kg to approximately 80 μg / kg, approximately 30 μg / kg to approximately 60 μg / kg, approximately 20 μg / kg to approximately 40 μg / kg, approximately 1 μg / kg to approximately 30 μg / kg, less than approximately 20 μg / kg of the subject's body weight, for example, approximately 0.01 μg / kg to approximately 10 μg / kg, or approximately 0.01 μg / kg to approximately 1 μg / kg, and / or approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg. For example, a suitable dosage typically provides and maintains a molecule containing or comprising an appropriate amount of annexin A5 protein (such as monomeric annexin A5 protein) to treat or prevent macular edema and / or retinal vein occlusion, or related conditions. A suitable dosage may aim to achieve and / or maintain levels of annexin A5 protein in the target plasma at levels higher than the naturally occurring physiological levels of annexin A5 in the plasma.

[0346] In other embodiments, the dose is administered over a long period, for example, the daily dose is delivered over a period of 30 minutes or more, for example, 1 hour or more, 2 hours or more, 3 hours or more, 4 hours or more, 5 hours or more, 6 hours or more, 7 hours or more, 8 hours or more, 9 hours or more, 10 hours or more, 11 hours or more, 12 hours or more, 13 hours or more, 14 hours or more, 15 hours or more, 16 hours or more, 17 hours or more, 18 hours or more, or up to 24 hours. In preferred embodiments, the daily dose is delivered by infusion over a period of time.

[0347] A treatment regimen may include, for example, a series of infusions of annexin A5 protein into the subject, or it may include administering one or more doses, for example, once, twice, three or four or more times a day, consisting of an amount of molecules containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) as defined in relation to the embodiments described above.

[0348] In some cases, a molecule containing or consisting of a therapeutically effective dose of annexin A5 protein (such as monomeric annexin A5 protein) may be optionally administered to a subject as two or more dose-escalating concentrations (i.e., escalating doses), in which case (i) all doses are therapeutic doses, or (ii) a dose less than the therapeutic dose (or two or more doses less than the therapeutic dose) is administered first, and the therapeutic dose is achieved by escalation. As one non-limiting example to illustrate escalating concentrations (i.e., dose increases), a therapeutically effective dose may be administered daily, starting with a dose less than the therapeutic dose, and each subsequent dose may be increased in specific or variable increments until the therapeutic dose is reached, at which point administration may be discontinued or continued (e.g., continuous therapeutic dose). In some embodiments, administration of the therapeutically effective dose may be by continuous infusion, and the dose may change over time (e.g., escalating). In some embodiments, combination therapy may also be administered.

[0349] In any case, the physician will determine the actual dosage of a molecule containing or comprising annexin A5 protein (such as monomeric annexin A5 protein) that is most suitable for any individual patient, and that dosage will vary depending on the patient's age, weight, and response. The dosages described above are examples of average cases. Naturally, there may be individual cases for which higher or lower dosage ranges are appropriate, and these are within the scope of the present invention.

[0350] For veterinary use, Annexin A5 protein is administered as a suitably acceptable formulation in accordance with standard veterinary practice, and the veterinarian determines the most appropriate administration regimen and route for the specific animal.

[0351] Treatment period: Therapeutic regimens containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) may be continued for a period of therapeutic benefit.

[0352] In one embodiment, the treatment may consist of a single dose of the Annexin A5 composition.

[0353] In another embodiment, multiple doses may be administered to the subject. For example, a dose consisting of an amount of molecules containing or comprising the annexin A5 protein (such as monomeric annexin A5 protein) as defined in the above embodiments may be administered once or more times per day, for example, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more times per day, preferably once per day.

[0354] In one embodiment, the dose of a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered at intervals of at least once every two days, at least once every three days, at least once every four days, at least once every five days, at least once every six days, at least once every seven days, at least once per week, at least once every two weeks, at least once every three weeks, at least once every four weeks, at least once per month, at least once every two months, at least once every three months, at least once every four months, at least once every five months, at least once every six months, at least once every seven months, at least once every eight months, at least once every nine months, at least once every ten months, at least once every eleven months, at least once every twelve months, at least once every two years, at least once every three years, at least once every four years, at least once every five years, or at least once every ten years.

[0355] In one embodiment, the dose of a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered over a period of one day or more, for example, for two or more consecutive days, three or more consecutive days, four or more consecutive days, five or more consecutive days, six or more consecutive days, seven or more consecutive days, eight or more consecutive days, nine or more consecutive days, ten or more consecutive days, eleven or more consecutive days, twelve or more consecutive days, thirteen or more consecutive days, two or more consecutive weeks, three or more consecutive weeks, four or more consecutive weeks, two or more consecutive months, three or more consecutive months, four or more consecutive months, five or more consecutive months, six or more consecutive months, seven or more consecutive months, eight or more consecutive months, nine or more consecutive months, ten or more consecutive months, eleven or more consecutive months Subsequently, it is administered daily for 12 consecutive months or more, 15 consecutive months or more, 18 consecutive months or more, 21 consecutive months or more, 24 consecutive months or more, 30 consecutive months or more, 3 consecutive years or more, 42 consecutive months or more, 4 consecutive years or more, 5 consecutive years or more, 6 consecutive years or more, 7 consecutive years or more, 8 consecutive years or more, 9 consecutive years or more, 10 consecutive years or more, 11 consecutive years or more, 12 consecutive years or more, 13 consecutive years or more, 14 consecutive years or more, 15 consecutive years or more, 16 consecutive years or more, 17 consecutive years or more, 18 consecutive years or more, 19 consecutive years or more, 20 consecutive years or more, 25 consecutive years or more, 30 consecutive years or more, 35 consecutive years or more, 40 consecutive years or more, or 50 consecutive years or more. In a preferred embodiment, a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered daily for a period of 6 consecutive days or more. In an alternative preferred embodiment, a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered daily for a period of seven or more consecutive days. In a more preferred embodiment, annexin A5 protein is administered daily for a period of six or seven consecutive days. In the most preferred embodiment, annexin A5 protein is administered daily for a period of six consecutive days. In an alternative most preferred embodiment, annexin A5 protein is administered daily for a period of seven consecutive days.

[0356] In one embodiment, a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered twice a day for a period of one week or more, for example, two weeks or more, three weeks or more, four weeks or more, two months or more, three months or more, four months or more, five months or more, six months or more, seven months or more, eight months or more, nine months or more, ten months or more, eleven months or more, twelve months or more, fifteen months or more, eighteen months or more, twenty-one months or more, twenty-four months or more, thirty months or more, three years or more, forty-two months or more, four years or more, five years or more, six years or more, seven years or more, eight years or more, nine years or more, ten years or more, eleven years or more, twelve years or more, thirty years or more, thirty years or more, thirty years or more, forty years or more, or fifteen years or more.

[0357] In one embodiment, a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered once a week for a period of one week or more, for example, two weeks or more, three weeks or more, four weeks or more, two months or more, three months or more, four months or more, five months or more, six months or more, seven months or more, eight months or more, nine months or more, ten months or more, eleven months or more, twelve months or more, fifteen months or more, eighteen months or more, twenty-one months or more, twenty-four months or more, thirty months or more, three years or more, forty-two months or more, four years or more, five years or more, six years or more, seven years or more, eight years or more, nine years or more, ten years or more, eleven years or more, twelve years or more, thirty years or more, thirty years or more, thirty years or more, forty years or more, or fifteen years or more.

[0358] In one embodiment, a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered twice a week or more for a period of one week or more, for example, two weeks or more, three weeks or more, four weeks or more, two months or more, three months or more, four months or more, five months or more, six months or more, seven months or more, eight months or more, nine months or more, ten months or more, eleven months or more, twelve months or more, fifteen months or more, eighteen months or more, twenty-one months or more, twenty-four months or more, thirty months or more, three years or more, forty-two months or more, four years or more, five years or more, six years or more, seven years or more, eight years or more, nine years or more, ten years or more, eleven years or more, twelve years or more, thirty years or more, thirty years or more, thirty years or more, forty years or more, or fifteen years or more.

[0359] In one embodiment, a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered at least twice a week for a period of one month or more, for example, two months or more, three months or more, four months or more, five months or more, six months or more, seven months or more, eight months or more, nine months or more, ten months or more, eleven months or more, twelve months or more, fifteen months or more, eighteen months or more, twenty-one months or more, twenty-four months or more, thirty months or more, three years or more, forty-two months or more, four years or more, five years or more, six years or more, seven years or more, eight years or more, nine years or more, ten years or more, eleven years or more, twelve years or more, thirty years or more, thirty years or more, thirty years or more, forty years or more, or fifteen years or more.

[0360] In one embodiment, a molecule containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) is administered at least twice a week for a period of one month or more, for example, two months or more, three months or more, four months or more, five months or more, six months or more, seven months or more, eight months or more, nine months or more, ten months or more, eleven months or more, twelve months or more, fifteen months or more, eighteen months or more, twenty-one months or more, twenty-four months or more, thirty months or more, three years or more, forty-two months or more, four years or more, five years or more, six years or more, seven years or more, eight years or more, nine years or more, ten years or more, eleven years or more, twelve years or more, thirty years or more, thirty years or more, thirty years or more, forty years or more, or fifteen years or more.

[0361] Combination therapy: All aspects of the present invention relate to combination therapies that utilize the administration of molecules containing or consisting of annexin A5 protein (such as monomeric annexin A5 protein) as co-therapy with one or more anti-VEGF agents.

[0362] Therefore, subjects treated by the present invention may be administered, in amounts and for durations that produce an overall therapeutic effect, a molecule containing or comprising annexin A5 protein (monomer annexin A5 protein) as part of co-therapy with one or more anti-VEGF agents. The molecule containing or comprising annexin A5 protein (monomer annexin A5 protein A5) can be combined with one or more anti-VEGF agents in a mixture, or by individual, simultaneous, or sequential administration (combination therapy).

[0363] Optionally, co-therapy using annexin A5 protein and anti-VEGF agents may be further supplemented with one or more additional agents. These additional agents may be administered in combination with the annexin A5 component and one or more anti-VEGF agents, either in a mixture with respect to either or both of the annexin A5 component and the anti-VEGF agents, or individually, simultaneously, or sequentially. To avoid misunderstanding, the other agents are not human annexin A5 or agents containing sequences of its biologically active fragments, more specifically, not dimeric annexin A5 (commonly referred to in the art as "dianexin"), nor are they PEGylated annexin A5.

[0364] The dosage and timing of administration of the annexin A5 component and the anti-VEGF component may, for example, produce additive or synergistic prophylactic and / or therapeutic effects. Furthermore, administration of molecules containing or comprising annexin A5 protein, such as monomeric annexin A5 protein (with or without a second or further agent), may be used as primary, e.g., first-line treatment, or as second-line treatment in response to one or more anti-VEGF agents and / or one or more additional agents. For example, annexin A5 may be administered as second-line treatment for subjects who have an inadequate response to previously administered treatments with anti-VEGF agents and / or one or more additional agents (i.e., previously administered treatments other than those using molecules containing or comprising annexin A5 protein, such as monomeric annexin A5 protein).

[0365] Accordingly, in one embodiment, the subject may be treated with therapy of a molecule containing or comprising annexin A5 protein, such as monomeric annexin A5 protein, in addition to one or more therapeutic or prophylactic interventions using an anti-VEGF agent and / or one or more additional agents. The subject may receive therapy of a molecule containing or comprising annexin A5 protein, such as monomeric annexin A5 protein, separately from, simultaneously with or consecutively with, one or more other therapeutic or prophylactic interventions using an anti-VEGF agent and / or one or more additional agents, or one or more additional therapeutic or prophylactic treatments using an anti-VEGF agent and / or one or more additional agents, in the same composition as them, or with separate formulations containing them.

[0366] In some embodiments of the present invention, one or more other therapeutic or prophylactic interventions, or one or more additional therapeutic or prophylactic treatments, include or consist of one or more treatments selected from the group consisting of anti-VEGF therapy (e.g., anti-VEGF therapy delivered to the eye by intravitreal injection), corticosteroids, NSAIDs, transsquamative vitrectomy, retinal laser photocoagulation, or anti-VEGF therapy enhanced with intravitreal corticosteroids and / or retinal laser photocoagulation.

[0367] Exemplary anti-VEGF therapies may include one or more of the following: ranibizumab, aflibercept, bevacizumab, brolucizumab, and / or falisimab.

[0368] Examples of corticosteroids include difluprednate, triamcinolone acetonide, and dexamethasone, which can be used as monotherapy and may or may not be combined with immunomodulatory therapy to achieve a more rapid resolution of edema.

[0369] Devices and kits for treatment: Pharmaceutical and veterinary compositions containing molecules comprising or derived from annexin A5 protein, such as monomeric annexin A5 protein, can be administered using medical devices. The devices may be designed with features such as portability, room temperature storage, and ease of use, so that in an emergency, they can be removed from medical facilities and other medical devices and used, for example, by untrained subjects or on-site emergency personnel. The devices may include one or more housings for storing pharmaceutical preparations containing or derived from annexin A5 protein, such as monomeric annexin A5 protein, and may be configured to deliver one or more unit doses of molecules containing or derived from annexin A5 protein, such as monomeric annexin A5 protein. The devices may further be configured to administer a second agent, such as an anti-VEGF agent, either as a single pharmaceutical composition also containing or derived from annexin A5 protein, such as monomeric annexin A5 protein, or as two separate compositions.

[0370] Pharmaceutical and veterinary compositions can be administered by syringe. Pharmaceutical or veterinary compositions can also be administered using needle-free subcutaneous injection devices, such as those disclosed in U.S. Patents 5,399,163, 5,383,851, 5,312,335, 5,064,413, 4,941,880, 4,790,824, or 4,596,556. Examples of well-known implants and modules include U.S. Patent No. 4,487,603 disclosing an implantable microinfusion pump for dispensing drugs at a controlled rate; U.S. Patent No. 4,486,194 disclosing a therapeutic device for administering drugs through the skin; U.S. Patent No. 4,447,233 disclosing a drug infusion pump for delivering drugs at a precise infusion rate; U.S. Patent No. 4,447,224 disclosing a variable flow rate implantable infusion device for continuous drug delivery; U.S. Patent No. 4,439,196 disclosing an osmotic drug delivery system having a multi-chamber compartment; and U.S. Patent No. 4,475,196 disclosing an osmotic drug delivery system. Many other devices, implants, delivery systems, and modules are also known.

[0371] Molecules containing or comprising annexin A5 protein, such as monomeric annexin A5 protein, can be provided in a kit. In one embodiment, the kit includes (a) a container for containing a composition comprising annexin A5 protein, and optionally (b) informational material. The informational material may be explanatory, instructional, marketing, or other material relating to the use of the agent for the therapeutic benefits of the method and / or the agent as described herein.

[0372] In embodiments, the kit also includes a second agent for treating the disorders described herein. For example, the kit includes a first container containing a composition comprising a molecule consisting of monomeric annexin A5 protein, and a second container containing the second agent. Optionally, the second agent is an anti-VEGF agent.

[0373] The informational materials of a kit are not limited to their form. In one embodiment, the informational materials may include information on the manufacture of the compound, the molecular weight, concentration, expiration date, batch, or place of manufacture of the compound. In one embodiment, the informational materials relate to a method of administering a molecule containing or consisting of annexin A5 protein, such as monomeric annexin A5 protein, in, for example, a preferred dose, dosage form, or mode of administration (e.g., the dose, dosage form, or mode of administration described herein) to treat a subject requiring treatment as described herein. In the case of a kit containing an anti-VEGF agent as a second agent, the kit may also include informational materials relating to a method of administering the anti-VEGF agent in, for example, a preferred dose, dosage form, or mode of administration (e.g., the dose, dosage form, or mode of administration described herein) to treat a subject requiring treatment as described herein. The information may be provided in various formats, including printed text, computer-readable material, video recordings, or audio recordings, or substantial material, such as information providing links or addresses to the Internet. In one embodiment, the information material is administered to the subject at doses of approximately 50 μg / kg to 100 μg / kg, approximately 40 μg / kg to 80 μg / kg, approximately 30 μg / kg to 60 μg / kg, approximately 20 μg / kg to 40 μg / kg, approximately 1 μg / kg to 30 μg / kg, and less than approximately 20 μg / kg per unit of body weight, for example, approximately 0.01 μg / kg to 10 μg / kg or approximately 0.01 μg / kg to 1 μg / kg, to monomeric annexin A5 protein. Instructions for use are provided for administering molecules containing or composed of annexin A5 protein, such as monomeric annexin A5 protein, and / or instructions for administering molecules containing or composed of annexin A5 protein, such as monomeric annexin A5 protein, to subjects in doses of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, less than approximately 2 mg, for example, approximately 1 mg, approximately 0.1 to 1.5 mg, 0.001 mg to approximately 1 mg, or approximately 0.001 mg to approximately 0.1 mg.

[0374] In addition to molecules containing or comprising annexin A5 protein, such as monomeric annexin A5 protein, the composition in the kit may include other components such as solvents or buffers, stabilizers, or preservatives. Molecules containing or comprising annexin A5 protein, such as monomeric annexin A5 protein, may be supplied in any form, e.g., liquid, dry, or lyophilized, preferably substantially pure and / or sterile. If molecules containing or comprising annexin A5 protein, such as monomeric annexin A5 protein, are supplied in a liquid solution, the liquid solution is preferably an aqueous solution. If molecules containing or comprising annexin A5 protein, such as monomeric annexin A5 protein, are supplied in a dry form, they are generally reconstituted by the addition of a suitable solvent. Solvents, such as sterile water or buffer solutions, may optionally be provided in the kit.

[0375] In the case of a kit containing an anti-VEGF agent as a second agent, the anti-VEGF agent may be supplied in any form, e.g., liquid, dry, or lyophilized, preferably substantially pure and / or sterile. If the anti-VEGF agent is supplied in a liquid solution, the liquid solution is preferably an aqueous solution. If the anti-VEGF agent is supplied in a dry form, it is generally reconstituted by the addition of a suitable solvent. The solvent, e.g., sterile water or buffer solution, may optionally be provided in the kit.

[0376] A kit may include one or more containers for one or more compositions containing a drug. In some embodiments, a kit includes separate containers, dividers, or compartments for the composition and informational material. For example, the composition may be contained in a bottle, vial, or syringe, and the informational material may be contained in a plastic sleeve or packet. In other embodiments, the separate elements of the kit are contained in a single, undivided container. For example, the composition is contained in a bottle, vial, or syringe with informational material attached in the form of a label. In some embodiments, a kit includes a plurality of individual containers (e.g., packs) each containing a drug in one or more unit dosage forms (e.g., dosage forms described herein). The containers may include a combined unit dose, for example, a unit containing both a molecule containing or consisting of annexin A5 protein, such as monomeric annexin A5 protein, and a second drug, such as an anti-VEGF agent, for example, in a desired ratio. For example, a kit may include a plurality of syringes, ampoules, foil packets, blister packs, or medical devices, each containing a single combined unit dose, for example. The kit container may be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or light-shielding.

[0377] The kit optionally includes one or more devices suitable for administering the compositions or each composition, such as syringes or other suitable delivery devices. The devices or each device may be provided pre-loaded with one or both of the compositions, or they may be empty but suitable for loading.

[0378] F. How to calculate the dosage of Annexin A5 In the eighth aspect, the present invention provides a method for calculating the dosage of annexin A5 necessary to treat conditions such as macular edema and / or RVO, the method comprising: A. A step to determine the number of annexin A5-positive cells in the subject, B. A step of determining the number of annexin A5 binding sites on annexin-positive cells in the subject, C. For example, a step of determining the number of annexin A5 molecules required to bind to the annexin A5 binding site by multiplying the value determined for A by the value determined for B, The process includes the step of determining the dosage from the value determined for DC.

[0379] The embodiments of the present invention discussed herein are applicable to the eighth aspect of the present invention unless otherwise stated.

[0380] In one embodiment, the method is From the value determined in C'.C, the molar concentration of the number of molecules required to bind to the annexin A5 binding site is determined, This further includes determining the dosage based on the values ​​determined for DC.

[0381] Preferably, D further includes determining the dose by multiplying the value determined for C' by the molar mass of annexin A5. Preferably, the molar mass of annexin A5 used is about 35,700 Da. If annexin A5 protein variants with different molecular weights are used, the values ​​used in calculation step D may be adjusted accordingly.

[0382] In one embodiment, annexin A5 is a molecule comprising monomeric annexin A5, for example, monomeric annexin A5 protein as further defined herein.

[0383] In one embodiment, the method is E. This includes administering Annexin A5 at the target dosage.

[0384] Preferably, the method is E. This includes administering a dose of Annexin A5 to the target patient to treat the condition.

[0385] In one embodiment, the method is an in vitro method. In another embodiment, steps A to D of the method are steps of an in vitro method.

[0386] In one embodiment, the condition is one or more selected from the list consisting of viral hemorrhagic fever (such as Ebola virus hemorrhagic fever or dengue virus hemorrhagic fever), cardiovascular condition (such as stroke, atherosclerosis, central retinal vein occlusion, myocardial infarction, stenosis, or restenosis), sickle cell disease, arthritis (such as osteoarthritis), and diabetes. Preferably, the condition is retinal vein occlusion, and more preferably central retinal vein occlusion.

[0387] In a ninth aspect, the present invention provides a method for calculating the dose of annexin A5 required to bind to the annexin A5 binding site in a subject, the method comprising: A. A step to determine the number of annexin A5-positive cells in the subject, B. A step to determine the number of annexin A5 binding sites on annexin A5-positive cells in the subject, C. For example, a step of determining the number of annexin A5 molecules required to bind to the annexin A5 binding site by multiplying the value determined for A by the value determined for B, The process includes the step of determining the dosage from the value determined for DC.

[0388] The embodiments of the present invention discussed herein are applicable to the ninth aspect of the present invention unless otherwise stated.

[0389] In one embodiment of the ninth aspect of the present invention, the method is: E. For prevention, preferably for the prevention of the condition, this includes administering a dose of annexin A5 to the target. * * * * * * * * * * * * * *

[0390] The present invention will now be described with reference to one or more non-limiting embodiments. [Examples]

[0391] The following embodiments are included to demonstrate specific embodiments of the present invention. Those skilled in the art will understand that the techniques disclosed in the following embodiments represent techniques that the inventors have found to function well in the implementation of the present invention and can therefore be considered to constitute a preferred mode for its implementation. However, those skilled in the art will understand that many modifications can be made to the specific embodiments disclosed in light of this disclosure, and that similar or comparable results can still be obtained without departing from the spirit and scope of the present invention.

[0392] Example 1 Probe construction and verification: Purified recombinant monomeric human annexin A5 protein (ANXV) was labeled with 800CW, a molecular imaging and contrast agent, to form ANXV-800CW. It was determined that approximately 40% of the ANXV present was labeled with 800CW (the remaining approximately 60% remained unlabeled monomeric human annexin A5 protein).

[0393] ANXV-800CW specifically binds to phosphatidylserine (PS), a key molecule in the pathogenesis of RVO.

[0394] A 66-year-old woman with a known history of systemic hypertension presented to the outpatient clinic. She had experienced decreased vision in her right eye over the past few weeks, which was determined to be caused by retinal branch vein occlusion (BRVO).

[0395] The PS assay was performed on blood samples collected before and after systemic administration of ANXV-800CW, and the presence of PS on fresh red blood cells was determined by flow cytometry using FITC-labeled annexin A5. These results showed an increased percentage of PS presence and PS availability compared to negative controls (Figure 1). This increase in PS externalization was observed throughout all blood sampling time points, demonstrating the stable availability of the ANXV-800CW target.

[0396] Evaluation of localization of annexin A5 administered intravenously to the site of retinal vein occlusion: Next, we evaluated the possibility of visualizing the coupling of ANXV-800CW at the site of retinal vein occlusion.

[0397] ANXV-800CW was administered intravenously as a single bolus at a dose of 1.0 mg, followed by near-infrared (NIR) fluorescence imaging using the Heidelberg Spectralis camera system at predetermined time points (up to 4 hours after injection).

[0398] Figure 2 shows the results of fluorescein angiography (FA), a standard treatment procedure. It shows the precise occlusion site (red arrow) and the damaged retinal region where bleeding is present (red oval). Upon performing NIR fluorescence imaging, the inventors observed a highly probable fluorescence signal for ANXV-800CW in the blood vessels and optic nerve located in the affected retinal region 5 minutes after administration. This intensity appeared to increase over time, as shown by the image created 229 minutes after injection.

[0399] In the surrounding retinal vessels identified by FA, ​​no fluorescence signal could be detected (see Figure 3). This indicates the absence of clear ANXV-800CW uptake in vessels not affected by BRVO.

[0400] These observations indicate site-specific accumulation of ANXV-800CW in retinal vessels, suggesting binding of annexin A5 at the occlusion site. The inventors interpret these results as demonstrating the efficacy of systemically administered annexin A5 (including labeled form ANXV-800CW) in preferentially accumulating at occlusion sites in subjects with exposed PS, its targets, and RVO.

[0401] Example 2 Background: The established standard for treating retinal vein occlusion (RVO) involves long-term, frequent direct reinjection of anti-VEGF into the eye via intravitreous injection (Nguyen et al., 2010, Ophthalmology, 117:2146-51; Campochiaro et al., 2010, Ophthalmology, 117:2387-94; Saint-Geniez et al, 2008, PLoS One., 3:e3554; Haydinger et al, 2023 (see above); Ip & Hendrick, 2018 (see above); Cox et al., 2021, J.Clin.Med., 10(5):981; Hattenbach et al., 2023, Ophthalmol Sci., 3(3):100302). However, anti-VEGF therapy raises concerns about ocular and systemic toxicity, requiring frequent and prolonged reinjections (Nguyen et al., 2010, Ophthalmology, 117:2146-51, Campochiaro et al., 2010, Ophthalmology, 117:2387-94, Saint-Geniez et al, 2008, PLoS One., 3:e3554). Potential complications associated with intravitreal injection of anti-VEGF therapy include infectious endophthalmitis, aseptic endophthalmitis, and brolucizumab-induced retinal vasculitis in particular (Cox et al., 2021 (see above)), and transient intraocular pressure (IOP) spikes (Levin et al., 2021, J Glaucoma, 30:1019-26).

[0402] Phosphatidylserine (PS) is a cell membrane lipid involved in the pathophysiology of retinal vein occlusion (RVO), diabetic retinopathy (DMR), and age-related macular degeneration (AMD). In RVO patients, PS is exposed on circulating red blood cells (RBCs) and contributes to their adhesion characteristics to microvascular endothelium, a feature implied to the cause of RVO. Recombinant human annexin A5 protein ANXV is a PS inhibitor that acts as an anti-inflammatory, cytoprotective, and anti-adhesion agent.

[0403] Methods: All previously untreated patients with early-onset RVO were eligible for this study. A total of 15 patients received intravenous ANXV as first-line treatment, with 6 patients receiving a daily dose of 2 mg, 3 patients receiving 4 mg, and 6 patients receiving 6 mg (n=6 for 2 mg, n=3 for 4 mg, and n=6 for 6 mg). ANXV doses were administered either over 5 consecutive days for some patients, or over 5 days within an 8-day period for others (primarily when access to consecutive treatment days within the treatment period was interrupted by weekends). All patients were followed for adverse events (AEs), laboratory parameters, pharmacokinetics (PK), anti-drug antibodies (ADAs), ophthalmic parameters, and IVT anti-VEGF requirements. Patients were studied (or will be studied, in the case of ongoing studies) for at least 4 months after initial treatment, and generally longer. At the time of writing, eight patients had been studied for at least four months (4m), two patients for at least three months (3m), two patients for at least two months (2m), one patient for at least one month (1m), and one patient was in the early stages of study for less than one month.

[0404] Results: A review of available safety and tolerable data from patients who received daily doses of either 2, 4, or 6 mg of ANXV and were followed up 10 days after their last dose of ANXV revealed no concerns.

[0405] Regarding the signal of efficacy in 8 out of 10 patients, information was available for at least 3 months post-treatment, and the data showed improvement or prevention of deterioration of visual acuity (BCVA) and, in some cases, reduction of retinal swelling, with or without anti-VEGF or with only one dose. Similarly, 10 out of 12 patients were considered to require no anti-VEGF treatment at all or only one dose, based on information available after 2 months. The decision to administer anti-VEGF is made by the patient's treating ophthalmologist, based on the presence of retinal swelling and decreased visual acuity.

[0406] [Table 2] TBC = Under review. As of the time of writing, the study of that specific patient is ongoing and has not yet reached the indicated time point.

[0407] Of the 15 RVO patients tested, 13 responded positively (improved or at least stabilized) to ANXV therapy. Only two patients (in groups B and D, both of whom received the lowest listed ANXV dose) responded and returned to conventional anti-VEGF therapy during the study period.

[0408] When visual acuity alone was evaluated using the BCVA test (acquired letters), the exemplary subjects showed the following outcomes, with particularly significant improvements from day 29 onwards.

[0409] [Table 3]

[0410] Compared to the expected number of patients who typically received an average of 4–6 anti-VEGF intravitreal injections within the first six months of RVO diagnosis (Light et al, 2020, Am. Acad. Ophthamol., 5(9):888), the number of anti-VEGF injections deemed necessary by treating ophthalmologists in the ANXV study group was zero or only one in 8 of the 10 patients followed for 3 months, and in 10 of the 12 patients followed for 2 months.

[0411] One such exemplary patient (Subject F, diagnosed with CRVO, and administered 2 mg of ANXV five times at a daily dose) was studied for seven months, and the study was terminated when the clinical endpoint was reached, demonstrating long-term and sustained improvement in visual acuity (BCVA), and a decrease in macular retinal thickness and macular volume.

[0412] [Table 4] ND: Not measured

[0413] Figure 4A shows the retinal sensitivity map obtained by microperimetry (MMP) of subject F, demonstrating that ANXV therapy resulted in clear improvement on day 15 post-treatment and complete recovery by day 120 post-treatment. Figure 4B shows imaging of blood flow in the retinal vascular system of subject F by fluorothane angiography, which also demonstrates clear improvement on day 15 post-treatment and complete recovery by day 29 or 120 post-treatment with ANXV therapy.

[0414] However, three of the patients who showed a signaling response to ANXV administration (subjects A, C, and E) were determined to require additional anti-VEGF treatment during the study period.

[0415] One of these subjects (Subject A) received aflibercept (Eylea®) approximately 32 days after joining the study. This subject was monitored by their ophthalmologist for a total of 16 months from the initial ANXV therapy and did not require any additional anti-VEGF treatment during the study period.

[0416] [Table 5] ND: Not measured

[0417] OCT (optical coherence tomography) measurements of macular retinal thickness in subject A were determined over a 16-month period following ANXV treatment and are shown in Figure 5. At 16 months, despite receiving only a single anti-VEGF treatment approximately 32 days after the start of the study, it was found that visual acuity remained stable at 20 / 60.

[0418] Conclusion: The data demonstrate a positive effect of ANXV therapy with a lower dose than the typical anti-VEGF requirement on the disease course. Prior to this study, the established standard of RVO treatment involved long-term, frequent reinfusions of anti-VEGF, associated with several potential adverse side effects. However, this study demonstrates that the need for anti-VEGF therapy can be significantly reduced when ANXV therapy is used. Where additional anti-VEGF treatment was needed, the required dose was still significantly reduced compared to the established standard, and generally, only a single anti-VEGF injection in combination with ANXV therapy was required over a period of more than 4 months (e.g., up to 16 months), without the need for multiple regular anti-VEGF injections, allowing subjects to reach the clinical endpoint and be freed from further cases.

[0419] These data demonstrate the ability of ANXV therapy to reduce the need and / or frequency of administration of anti-VEGF agents. The inventors conclude that ANXV can be used for the prevention or treatment of VEGF-related diseases, disorders or conditions in subjects, and / or for the prevention or treatment of diseases, disorders or conditions in subjects accompanied by one or more administrations of anti-VEGF agents to subjects. The inventors further conclude that ANXV can be used to enhance anti-VEGF therapy and / or to reduce complications associated with the administration of anti-VEGF agents.

[0420] Example 3 As discussed above, the evidence suggests that several inflammatory cytokines, including IL-6, TNF-α, IL-1β, and ICAM-1, may act to disrupt BRB integrity and contribute to the development of macular edema and / or retinal vein occlusion (Haydinger et al., 2023 (see above), Park et al., 2019 (see above), Noma et al., 2020 (see above)).

[0421] Here, the inventors report experiments suggesting that annexin A5 may block the effects of such inflammatory cytokines.

[0422] Methods: Chondrocytes were cultured with IL-1β (10 ng / mL) or TNF-α (10 ng / mL) with / without purified recombinant human annexin A5 (ANXA5) (100 ng / mL, 5 μg / mL, 20 μg / mL). Cells were harvested at 3, 6, 12, and 18 hours. Cell viability was determined by flow cytometry using propidium iodide (PI).

[0423] IL-6 expression was measured in IL-1β-stimulated cells. Cells were stimulated with 10 ng of IL-1β and treated with different concentrations of ANXA5 (0.1 μg, 5 μg, and 20 μg / mL) for 6 hours. Gene expression was measured by qPCR using the SYBR Green reaction kit. Gene expression was normalized to GAPDH, and relative gene expression was measured using 2ΔΔCt.

[0424] Results: The results indicate that treatment with Annexin A5 reduces cell death in this ex vivo study.

[0425] Figure 6 shows that in a human model, cell death promoted by TNFα application was inhibited by annexin A5 treatment, and the cell viability of annexin A5-treated cells was similar to that of controls without TNFα treatment.

[0426] As shown in Figure 7, similar results were observed with IL-1β. Annexin A5 was shown to inhibit cell death in a dose-dependent manner, with treatment at doses of 5 μg and 20 μg resulting in cell viability comparable to that of a control without IL-1β treatment. Furthermore, Figure 7 shows that treatment with annexin A5 protein alone did not affect cell viability compared to a control without IL-1β treatment.

[0427] In addition to inhibiting cell death, treatment with annexin A5 also reduces important biomarkers.

[0428] As shown in Figure 8, treatment with Annexin A5 reduces ICAM1 expression compared to cells in which cell death and inflammation are promoted using TNFα, as well as to untreated controls.

[0429] As shown in Figure 9, treatment with Annexin A5 dose-dependently reduces IL-6 expression compared to cells induced with IL-1β and untreated controls. In addition, Figure 9 shows that treatment with Annexin A5 alone does not affect IL-6 expression.

[0430] Conclusion: Annexin A5 is involved in the dysregulation of BRB integrity and is demonstrated to inhibit the production and / or effects of several inflammatory cytokines, including IL-6, TNF-α, IL-1β, and ICAM-1, which contribute to the development of macular edema and / or retinal vein occlusion. * * * * * * * * * * * * * *

[0431] Any use, method, or composition described herein is considered to be applicable to any other use, method, or composition described herein.

[0432] In the claims and / or herein, the use of the words "a" or "an" in conjunction with the term "including" may mean "one," but also coincide with the meanings of "one or more," "at least one," and "one or more."

[0433] Unless otherwise stated, the term “about” as used herein may mean a range of ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned.

[0434] While the above description illustrates various embodiments of the present invention and numerous specific details thereof, it should be understood that it is given as an example and not an limitation. Many substitutions, modifications, additions, and / or rearrangements may be made within the scope of the present invention without departing from the spirit of the invention, and the present invention includes all such substitutions, modifications, additions, and / or rearrangements. * * * * * * * * * * * * * *

[0435] All compositions and / or methods disclosed and claimed herein can be prepared and performed without excessive experimentation in light of this disclosure. While the compositions and methods of the present invention have been described in relation to preferred embodiments, it will be apparent to those skilled in the art that the compositions and / or methods, as well as the steps or order of steps of the methods, may be modified without departing from the concept, spirit and scope of the invention. More specifically, certain chemically and physiologically relevant agents may be used in place of the agents described herein, and it will be apparent that the same or similar results can be achieved. All such similar substitutions and modifications, which will be apparent to those skilled in the art, are considered to fall within the spirit, scope and concept of the invention as defined by the appended claims.

Claims

1. A molecule comprising or consisting of annexin A5 protein for use in the prevention or treatment of macular edema or related conditions in a subject, wherein such use is in combination with an anti-vascular endothelial growth factor (anti-VEGF) agent.

2. A molecule comprising or consisting of annexin A5 protein for use according to claim 1, wherein the dose of the molecule is administered to the subject systemically, preferably intravenously or by choroidal injection.

3. A molecule comprising or consisting of annexin A5 protein for use according to claim 1 or 2, wherein the dose of the molecule is administered to the subject daily for two or more consecutive days.

4. A molecule comprising or consisting of annexin A5 protein for use according to claim 3, wherein the dose of the molecule is administered to the subject daily for three, four, five, six, seven or more consecutive days.

5. The daily dose of the molecule administered to the subject is a) A total daily dose of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, b) Within a range selected from the group consisting of approximately 50-100 μg, approximately 40-80 μg, approximately 30-60 μg, approximately 20-40 μg, or less than approximately 20 μg per kg of body weight of the subject. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 4.

6. The daily dose of the molecule administered to the subject is a) A total daily dose of 0.1 to 1.5 mg, or b) Within the range of 1 μg to 30 μg per kg of body weight of the subject, A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 4.

7. A molecule comprising or consisting of annexin A5 protein for use according to claim 1, wherein the subject has, is suspected of having, or is at risk of having retinal vein occlusion (RVO).

8. A molecule comprising or comprising annexin A5 protein for use according to any one of claims 1 to 7, wherein the condition associated with macular edema is selected from the group consisting of RVO, dysregulation of the blood-retinal barrier (BRB), e.g., dysregulation of the medial BRB and / or lateral BRB, dysfunction of the barrier between the vitreous and ciliary vessels, eye diseases associated with macular edema, e.g., diabetic retinopathy, retinal vascular occlusion, uveitis, post-cataract surgery inflammation (pseudophakic macular edema or Irving-Gas syndrome), retinal dystrophy, drug reaction, intraocular tumor, serous central chorioretinopathy, radiation retinopathy, and other retinal vascular abnormalities including retinal microaneurysms and retinal telangiectasia, changes in the subretinal neovascular membrane observed in conditions such as age-related macular degeneration (AMD), metamorphopsia, micropsia, blurred vision, central scotoma, and decreased contrast or color sensitivity.

9. A molecule containing or comprising the annexin A5 protein for use in the prevention or treatment of retinal vein occlusion (RVO) or related conditions in a subject, The molecule comprising monomeric annexin A5 protein is administered to the subject daily for two or more consecutive days, either systemically or by choroidal injection. The aforementioned use is in combination with anti-vascular endothelial growth factor (anti-VEGF) agents. A molecule containing or consisting of annexin A5 protein for use.

10. A molecule comprising or consisting of annexin A5 protein for use according to claim 9, wherein the dose of the molecule is administered intravenously to the subject daily for two or more consecutive days.

11. A molecule comprising or consisting of annexin A5 protein for use according to claim 9 or 10, wherein the dose of the molecule is administered to the subject systemically, preferably intravenously or by choroidal injection, daily for three, four, five, six, or seven days or more consecutively.

12. The daily dose of the molecule administered to the subject is, systemically, preferably by intravenous or choroidal injection. a) A total daily dose of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg, b) Within a range selected from the group consisting of approximately 50-100 μg, approximately 40-80 μg, approximately 30-60 μg, approximately 20-40 μg, or less than approximately 20 μg per kg of body weight of the subject. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 9 to 11.

13. The daily dose of the molecule administered to the subject systemically, preferably by intravenous or choroidal injection, a) A total daily dose of 0.1 to 1.5 mg, or b) Within the range of 1 μg to 30 μg per kg of body weight of the subject, A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 9 to 11.

14. The use described above is for the prevention or treatment of a condition in a subject who has, is suspected of having, or is at risk of having retinal vein occlusion (RVO), A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 9 to 13, wherein the condition is selected from the group consisting of macular edema, retinal ischemia, iris neovascularization (optionally, iris neovascularization characterized by an increase in cytokines such as an increase in VEGF), retinal neovascularization (optionally, retinal neovascularization characterized by an increase in cytokines such as an increase in VEGF), neovascular glaucoma, vitreous hemorrhage, rubeosis, and retinal detachment.

15. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 14, wherein the subject is a human adult and / or has a body weight of about 50 kg to about 150 kg.

16. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 7 to 15, wherein the RVO is selected from central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), or hemiretinal vein occlusion (HRVO).

17. A molecule comprising or consisting of an annexin A5 protein for use according to any one of claims 1 to 16, wherein the annexin A5 protein is a monomeric annexin A5 protein.

18. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 17, wherein the molecule comprises monomeric annexin A5 protein.

19. The aforementioned annexin A5 protein (optionally, monomeric annexin A5 protein) a. Proteins that contain, are essentially derived from, or consist of the sequence of human annexin A5 protein (human monomeric annexin A5 protein and / or optionally, proteins containing or consisting of the sequence of SEQ ID NO: 1, with or without N-terminal methionine), b. Recombinant human monomer annexin A5 protein, etc. c. Mammalian orthologues of human annexin A5 protein, such as human monomeric annexin A5 protein. d. Alleles or genetic variants of a), b), or c), e. Functional analogues or variants of human annexin A5 protein (such as monomeric annexin A5 protein) having the sequence defined by SEQ ID NO: 1, either containing or not containing an N-terminal methionine, wherein the protein contains or consists of 50%, 60%, 70%, 75%, for example, more than 80%, 85%, 90%, or more more than 95%, or 99% of the amino acid sequence of identity. f. Any biologically active fragment from a to e) A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 18, selected from one or more of the above.

20. A composition comprising an annexin A5 protein (such as monomeric annexin A5 protein) or a molecule comprising the same for use according to any one of claims 1 to 19, the composition also comprising one or more anti-vascular endothelial growth factor (anti-VEGF) agents.

21. The composition for use according to claim 20, wherein the composition is a composition in a unit dose.

22. The unit dose composition is a) A total dose of approximately 5 mg, approximately 4 mg, approximately 3 mg, approximately 2 mg, or less than approximately 2 mg of the molecule containing or consisting of annexin A5 protein, b) The molecule containing or consisting of annexin A5 protein in a total dose of 0.1 to 1.5 mg. A unit dose composition for use according to claim 21, comprising:

23. The composition for use according to any one of claims 20 to 22, wherein the composition is a pharmaceutical composition.

24. The composition for use according to any one of claims 20 to 23, wherein the composition further comprises one or more pharmaceutically acceptable carriers.

25. The composition for use according to claim 24, wherein the composition comprises a molecule containing or consisting of annexin A5 protein and one or more pharmaceutically acceptable carriers.

26. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 19, wherein the use is as a combination therapy using an anti-VEGF agent and optionally another agent, the composition for use according to any one of claims 20 to 23.

27. When used as a combination therapy with an anti-VEGF agent and, optionally, another drug, (a) The molecule containing or consisting of the annexin A5 protein is used as a first-line treatment with or without the anti-VEGF agent and / or one or more other agents, or (b) After treatment with the anti-VEGF agent and / or one or more other agents has already been initiated by at least one administration thereof to the subject, the molecule comprising or consisting of the annexin A5 protein is used. A composition comprising a molecule containing or consisting of annexin A5 protein for use according to any one of claims 1 to 19 or 26, or a molecule for use according to any one of claims 20 to 26.

28. When used as a combination therapy with an anti-VEGF agent and, optionally, another drug, The subject is administered, in the same composition as the anti-VEGF agent, separately, simultaneously with, or sequentially with, a separate formulation containing the annexin A5 protein. Optionally, the subject may be administered, in the same composition as one or more other drugs, separately from them, simultaneously with or sequentially with a separate formulation containing them. A composition comprising or consisting of an annexin A5 protein for use according to any one of claims 1 to 19, 26, or 27, or a molecule for use according to any one of claims 20 to 27.

29. The period between consecutive administrations of the anti-VEGF agent after the administration of the molecule or composition to the subject is longer than 28 days, for example, longer than a period selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 months, or longer than 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5 years. Optionally, the anti-VEGF therapy may be maintained by repeated administration to the subject for a period of approximately up to or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months, or approximately up to or at least 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 years. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 19, 26, 27, or 28, or a composition for use according to any one of claims 20 to 28.

30. The aforementioned anti-VEGF agent is applied to the subject, i. Within a period of 12 months from the start of treatment with the molecule or composition, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than 1 time, ii. Within a period of 10 months from the start of treatment with the molecule or composition, 10, 9, 8, 7, 6, 5, 4, 3, 2 or less than 1 time, iii. Within a period of 8 months from the start of treatment with the molecule or composition, 8, 7, 6, 5, 4, 3, 2 or less than 1 dose, and / or iv. Within a period of six months from the start of treatment with the molecule or composition, 6, 5, 4, 3, 2, or less than 1 dose. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 19 or 26 to 29, or a composition for use according to any one of claims 20 to 29, to be administered.

31. a) After the molecule or composition is administered to the subject, the frequency of administration of the anti-VEGF agent is reduced compared to a preferred control, or b) After administration of the molecule or composition to the subject, the subsequent administration of the anti-VEGF agent is delayed (e.g., prevented) compared to a suitable control. A molecule comprising or consisting of annexin A5 protein for use according to any one of claims 1 to 19 or 26 to 30, or a composition for use according to any one of claims 20 to 30.

32. a) Compared to a preferred control, the frequency of administration of the anti-VEGF agent is reduced by 1, 2, 3 or 4 weeks, or by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, or more than 1 year, and / or b) Based on the presence or absence of one or more symptoms associated with macular edema or RVO, or related conditions, the need for subsequent administration of the anti-VEGF agent is evaluated, and optionally, if the symptoms are present, xvii. Visual acuity loss, optionally evaluated by best corrected visual acuity (BCVA). xviiii. Increased macular retinal thickness, iii. Increase in macular volume, and xx. Increase in retinal non-perfusion areas (RANPs) A molecule or composition comprising annexin A5 for use according to claim 31, selected from the group consisting of one or more of the following.

33. A molecule or composition comprising annexin A5 for use according to claim 31 or 32, wherein the preferred control is a subject that has not been treated with a molecule comprising or comprising annexin A5 protein.

34. a) Prevention or treatment of VEGF-related diseases, disorders or conditions in the subject, and / or prevention or treatment of said diseases, disorders or conditions in the subject, accompanied by one or more administrations of anti-VEGF agents to the subject. b) Enhancement of anti-VEGF therapy, and / or c) Reduction of complications associated with the administration of anti-VEGF agents. A molecule containing or consisting of annexin A5 protein for use in [a specific application / procedure].

35. Refer to this specification for a molecule comprising or consisting of the annexin A5 protein, or a composition comprising the said molecule, or a use or method thereof, for use substantially described herein.

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