Iga for treatment and prevention of dry eyes

Abstract

The present disclosure refers to Immunoglobulin A (IgA) for use in a method for treating or preventing dry eyes in a patient wherein said Immunoglobulin A (IgA) is obtained from plasma, blood plasma or plasma fractions, and wherein said Immunoglobulin A (IgA) is administered to said patient in an effective dosage. In particular, the disclosure pertains to the use in a treatment or prevention of dry eye disease and the respective complications like infections. Further, the disclosure refers to IgA obtained from plasma, blood plasma or plasma fractions and a pharmaceutical composition comprising such IgA.

Description

[0001] PreviPharma GmbH October 30, 2025

[0002] PREV75185PC

[0003] IgA for treatment and prevention of dry eyes

[0004] The present invention refers to Immunoglobulin A (IgA) for use in a method for treating or preventing dry eyes in a person or patient wherein said Immunoglobulin A (IgA) is obtained from plasma, blood plasma or plasma fractions, and wherein said Immunoglobulin A (IgA) is administered to said patient in an effective dosage. In particular, the invention pertains to the use in a treatment or prevention of dry eye disease and the respective complications like infections. Further, the invention refers to IgA obtained from plasma, blood plasma or plasma fractions and a pharmaceutical composition comprising such IgA.

[0005] The present invention is set in the context of a treatment or prevention of dry eyes in a person or patient, in particular in the context of the dry eyes disease (DED). The dry eyes disease, also known as dry eyes syndrome (DES) or keratoconjunctivitis sicca (KCS) involves complex ocular surface issues that lead to discomfort and visual disturbances. [Huang R, et al., Int Ophthalmol. 2022 Oct;42(10):3253-3272], DES and DED refer to a common ocular condition characterized by inadequate lubrication and moisture on the eye's surface. This condition can cause various symptoms, including dryness, irritation, burning, redness, and blurred vision.

[0006] The Tear Film & Ocular Surface Society (TFOS) Dry Eye Workshop (DEWS) II describes dry eye disease (DED) as a complex condition affecting the ocular surface, marked by an imbalance in the tear film and associated with various symptoms. This condition involves instability and high osmolarity of the tear film, inflammation and damage to the ocular surface, and abnormalities in sensory nerves, all of which contribute to its development [Craig JP, et al., Ocul Surf, 2017;15:802-12],

[0007] Dry eye disease is marked by tear film instability, either from insufficient tear production or from poor tear quality, leading to increased evaporation. It is primarily categorized into two types: (1 ) aqueous production deficient dry eye and (2) evaporative dry eye. Both types cause ocular surface damage and discomfort [King- Smith PE, et al., Invest Ophthalmol Vis Sci. 2000 ;41 (11 ):3348-59],

[0008] The tear film, about 2 to 5.5 pm thick over the cornea, consists of three main layers [Peng CC, et al., Adv Colloid Interface Sci. 2014 ;206:250-64], The outermost lipid layer, produced by the meibomian glands, reduces tear evaporation [Zhou L, et al., Prog Retin Eye Res. 2012 Nov;31 (6):527-50], The thick aqueous middle layer, from the lacrimal and accessory lacrimal glands, contains water, electrolytes, peptides, and proteins. The basal mucin layer, secreted by conjunctival goblet cells, enhances tear film stability on the corneal epithelium by regulating surface tension [Mantelli F,et al., Curr Opin Allergy Clin Immunol. 2013 Oct; 13(5): 563-8],

[0009] Dry eye disease (DED) is a major public health concern, impacting approximately 344 million people globally, more than 20 million individuals in the United States alone and approximately 15 million diagnosed patients in the five largest European countries [https: / / www.novaliq.com / press-releases / 2024 / 07 / 29 / novaliq-receives-positive- chmp-opinion-for-vevizye-in-dry-eye-disease / ]. Dry eye disease (DED) is typically regarded as an adult condition, with prevalence estimates ranging from 5% to 30% among individuals aged 50 and older, and it is more common in women [Stapleton F, et al., Ocul Surf 2017; 15:334-65],

[0010] Dry eye disease (DED) can cause a range of symptoms from mild to severe. Common issues include stinging, burning, or a gritty sensation in the eyes, and paradoxically, excessive tearing due to dryness-induced irritation.

[0011] Pain may be sharp or dull, localized to the eye or surrounding areas. Redness is often exacerbated by rebound effects from vasoconstrictors in some over-the-counter eye drops. Blurry vision, glare, and haloes around lights are frequent complaints. Other symptoms include fluctuating vision, eyelid heaviness or twitching, excessive blinking, and discomfort for contact lens users. Some individuals also experience tired eyes and difficulty crying [Mohamed HB.et al., Eur J Pharm Sci. 2022 ; 175: 106206],

[0012] Multiple factors can contribute to the development of dry eye disease (DED). These include systemic medications (e.g., antihistamines, diuretics, corticosteroids), topical treatments, skin conditions around the eyelids (e.g., rosacea, eczema), meibomian gland dysfunction, and ophthalmic surgery. Other causes include chemical burns, ocular allergies, decreased androgen levels, excessive or insufficient vitamin intake, prolonged contact lens use, and neurotrophic cornea conditions. Systemic diseases (e.g., Sjogren's syndrome, diabetes), environmental factors (e.g., smoke, high temperature), and lifestyle factors (e.g., poor sleep, unhealthy diet, aging) also play a role. [Qian L,et.al., PLoS One. 2022 Aug 19; 17(8)].

[0013] Existing treatment strategies focus on alleviating symptoms, restoring the ocular surface and tear film, improving visual perception and quality of life, and / or addressing underlying causes. The treatment options have evolved significantly, and several new therapies and advancements have emerged recently. In this regard follows a summary of the latest treatments and their associated side effects.

[0014] The 0.05% cyclosporine was approved by the FDA in 2003 for treating inflammation and androgen deficiency-induced dry eye syndrome. While these treatments can help keep the eyes moist, reduce tear evaporation, and alleviate dryness of the ocular surface, their use may also elevate the risk of toxic and irritating side effects and potentially lead to drug-resistant infections. Consequently, non-steroidal antiinflammatory drugs (NSAIDs) are being used more frequently as an alternative to steroids for managing dry eye disease.

[0015] The nerve growth factor (NGF), discovered in the 1950s, is vital for the growth, maintenance and survival of certain neurons and acts as a signalling molecule through its receptors. NGF and its receptors have been found in rat lacrimal glands and human tears [Lee et al., Am J Ophthalmol, 2005, 139 (6), 965-71 ], It promotes corneal epithelial cell proliferation and differentiation, aiding in ocular surface maintenance and corneal wound healing [Lambiase et al., Invest Ophthalmol Vis Sci, 2009 ,50 (10), 4622-30],

[0016] NGF eye drops increase tear production and goblet cell density in dry eye models [Coassin M. et al., Graefes Arch Clin Exp Ophthalmol, 2005, 243 (2), 151 -5], NGF also helps conjunctival epithelial cells become goblet cells, which secrete mucin to protect the ocular surface [Lambiase et al., Invest Ophthalmol Vis Sci, 2009, ,50 (10), 4622-30], Studies reported that NGF administration enhances ocular surface sensitivity, reduces inflammation, and regulates tear production, underscoring NGF's importance in dry eye treatment [Lin_H et al. Saudi Journal of Ophthalmology, 2014 28(3): p. 173-181 ],

[0017] The U.S. Food and Drug Administration (FDA) granted approval for Xiidra [Lifitegrast ophthalmic solution] to treat the signs and symptoms of dry eye disease. This makes Xiidra the first drug in a novel class called lymphocyte function-associated antigen 1 (LFA-1 ) antagonists to be approved by the FDA for this condition. The medication is manufactured by Shire US Inc. in Lexington, Massachusetts. Lifitegrast interferes with the interaction between cell surface proteins LFA-1 and intercellular adhesion molecule-1 (ICAM-1 ), which is thought to reduce T-cell-mediated inflammation in dry eye disease (DED). Lifitegrast (Xiidra) is generally well-tolerated, but like all medications, it can cause side effects. The most commonly reported side effects include eye irritation, taste disturbances, headache, conjunctival hyperemia, reduced visual acuity, dry eye symptoms worsening, allergic reactions [Donnenfeld, E.D, et al., Expert Opin. Pharmacother. 2017, 18, 1517-1524],

[0018] The FDA approved 0.05% cyclosporine A (CsA) eye drops (Restasis) for the treatment of dry eye in 2002. Topical application of CsA can enhance tear production, likely through the local release of parasympathetic neurotransmitters. CsA is an immunosuppressant that works by forming a complex with cyclophilin, inhibiting the calcineurin-phosphatase pathway and thereby decreasing the transcription of T-cell- activating cytokines like interleukin-2 (IL-2) [Periman LM,et al., Clin Ophthalmol. 2020 Dec 2; 14:4187-4200]

[0019] Loteprednol etabonate 0.25% suspension, marketed as Lotemax, is primarily approved by the FDA for the treatment of inflammation and pain associated with ocular surgery and allergic conjunctivitis, not specifically for dry eye disease. However, it is sometimes used off-label to manage dry eye disease, particularly when inflammation is a significant component [Noriaki N. et al., Adv Drug Delivery Reviews 191 (2022) 114582],

[0020] Many compounds and drugs are used as a complementary treatment for dry eye disease with potential side effects. Polyvinyl alcohol, povidone, hydroxypropyl guar, cellulose derivatives, and hyaluronic acid artificial tears increase tear film stability, educe ocular surface stress, improve contrast sensitivity and the optical quality of the surface.

[0021] Omega-3 and omega-6 fatty acids are crucial for maintaining the health and stability of the ocular surface. Omega-3 fatty acids exert their effects by inhibiting pro- inflammatory eicosanoids and decreasing cytokine levels through their antiinflammatory properties. It has been used as complementary treatment for dry eye disease. They are believed to help reduce inflammation and improve the quality of tears [Barabino, S.et al., Cornea 2003, 22, 97-101 ],

[0022] Tears have antimicrobial, nourishing, mechanical, and optical properties. They contain components such as growth factors, fibronectin, and vitamins to support proliferation, migration, and differentiation of the corneal and conjunctival epithelium. A lack of these epitheliotrophic factors — for example, in dry eye, can result in severe ocular surface disorders such as persistent epithelial defects. Persistent corneal epithelial defects present a very challenging problem to anterior segment surgeons. Autologous serum tears had been demonstrated to be beneficial in the treatment of persistent corneal epithelial defects. Several studies were published showing the beneficial effect of autologous serum (tears) [Young AL et al.: Eye e (2004) 18, 609- 614],

[0023] Immunoglobulin therapy, particularly the use of topical immunoglobulins, is an emerging approach for managing dry eye disease, especially in cases where inflammation and immune system dysfunction play significant roles. Immunoglobulins can bind to and neutralize inflammatory mediators, reducing overall inflammation on the ocular surface. Intravenous immunoglobulin (IVIG) used for more severe systemic conditions that impact the eyes for severe or systemic forms of immune-mediated dry eye disease [Kate A, et al., Indian J Ophthalmol. 2023 Apr;71 (4): 1176-1189].

[0024] Secretory IgA (slgA) acts as a primary defense at mucosal surfaces, including the ocular mucosa. This encompasses the ocular surface itself (the conjunctiva and cornea) and its associated mucosal structures, such as the lacrimal gland and the lacrimal drainage system, which together function as a cohesive anatomical and functional unit [Knop E, et al., Immune Response and the Eye. 2007;36-49],

[0025] Secretory IgA (slgA) plays a critical role in maintaining ocular surface health by providing immune protection and modulating inflammation. In dry eye disease, a deficiency or dysfunction of slgA can contribute to increased inflammation and symptoms. Understanding and potentially enhancing slgA levels through various therapeutic approaches could offer additional strategies for managing dry eye disease [de Paiva, C.S., et al., Mucosal Immunol 15, 1143-1157 (2022)] slgA is essential for tear film stability, binding pathogens and preventing their adherence to the corneal epithelium. In dry eye disease, where the tear film is compromised, IgA levels and functionality can decrease, potentially reflecting a weakened immune response and heightened risk of ocular inflammation.

[0026] The treatment of ocular disorders with biological fluids has long been advocated [Hirschberg J. The History of Ophthalmology. Volume 1 : Antiquity. JP Wayenborgh Verlag: Bonn, 1985; Fox Rl, Chan R, Michelson JB et al. Beneficial effect of artificial tears made with autologous serum in patients with keratoconjunctivitis sicca. Arthr Rheum 1984; 27: 459^61 ],

[0027] Autologous serum has been used to treat dry eye syndrome for many years. Currently the majority of scientists believe that several growth factors, vitamins, fibronectin and other components are important for corneal and conjunctival integrity. Serum eye drops are usually prepared as an unpreserved blood solution. The serum is by nature well tolerated and its biochemical properties are somewhat similar to natural tears. Autologous serum eye drops have been reported to be effective for the treatment of severe dry eye-related ocular surface disorders (Sjogren’s syndrome), and also other entities such as superior limbic keratoconjunctivitis, graft-versus-host disease, Stevens-Johnson syndrome, ocular cicatricial pemphigoid, recurrent or persistent corneal erosions, neurotrophic keratopathy, Mooren’s ulcer, aniridic keratopathy, filtering blebs after trabeculectomy, and post-keratorefractive surgery [Quinto GG et al.:, Arq Bras Oftalmol. 2008;71 (6 Supl):47-54],

[0028] Conventional therapeutic options include intensive artificial tear supplements, punctual occlusion, contact lenses, and appropriate management of adnexal disease. The most frequent therapy utilized to treat ocular surface disorders is artificial tear eye drops. However, none of the commercially available artificial tear preparations includes essential tear components such as growth factors, vitamins, or immunoglobulin G (IgG). Another drawback of artificial tears is the fact that they often contain preservatives, stabilizers, and other additives, which potentially induce toxic or allergic reactions. Despite maximal conventional therapy, there is a cohort of patients who still have persistent symptoms and signs. Such patients may have more serious ocular surface disorders with significant visual impairment and disability.

[0029] Table 1. Comparison of the biochemical properties of normal human tears and serum [Geerling G, Maclennan S, Hartwig D. Autologous serum eye drops for ocular surface disorders. Br J Ophthalmology. 2004;88(11 ): 1467-74],

[0030] EGF= epithelial growth factor; TGF= transforming growth factor Especially in patients with an infection on the eye, the IgA will be increased. In chronical infection or in the dry eye disease beside the lack of liquid, IgA as the first defense line is missing.

[0031] Despite different research and approaches, there remains a need for safer and more effective treatment regimens for dry eyes in patients. There is still an unmet need for a treatment or prevention of dry eyes, restoring the eye morphology and functioning such as tear production and secretion long term and without the potential risk of side effects.

[0032] Surprisingly, it was found that the substitution of Immunoglobulin A (IgA), especially IgA derived from body fluids, in particular from plasma pools, which represents a high number of different antibody specificities, results in an improved treatment and prevention of dry eyes and accelerates healing of corneal infections and defects.

[0033] Summary of the Invention

[0034] Accordingly, the invention relates to Immunoglobulin A (IgA) for use in a method for treating or preventing dry eyes in a patient wherein said IgA is obtained from plasma, blood plasma or plasma fractions, and wherein said IgA is administered to said patient in an effective dosage. In particular, the invention pertains to the use in a treatment or prevention of dry eye disease and the respective complications like infections. Further, the invention refers to IgA obtained from plasma, blood plasma or plasma fractions and a pharmaceutical composition comprising such IgA.

[0035] Detailed Description of the Invention

[0036] The present application discloses Immunoglobulin A (IgA) for use in a method for treating or preventing dry eyes in a patient wherein said IgA is obtained from plasma, blood plasma or plasma fractions, and wherein said IgA is administered to said patient in an effective dosage.

[0037] In a first aspect, the invention relates to Immunoglobulin A (IgA) for use in a method for treating or preventing dry eyes in a patient wherein said IgA is obtained from plasma, blood plasma or plasma fractions at a purity of at least 50%, in particular wherein impurities mainly consist of immunoglobulins other than IgA, albumin, lysozyme, transferrin, ferritin, fetuin and / or proteases, and wherein said IgA is administered to said patient in an effective dosage.

[0038] In the context of the application, the term “dry eyes” is to be understood in a broad way, referring to the condition wherein no adequate lubrication for a patient’s eyes is provided. This may be due to various reasons and a short term or long-term condition, or a risk due to a specific circumstance or exposition. As used in the context of the present invention, the term “patient” may be understood in the broadest sense as any living being, which is preferably any animal, more preferably a mammal including human, in particular a human being. The patient may show symptoms of dry eyes or may be healthy, but at risk of developing dry eyes.

[0039] Immunoglobulin A (IgA) is to be understood in a broad sense, in particular as an IgA fraction i.e. a selected stake of the IgA pool, of a body fluid, in particular plasma, blood plasma or plasma fractions. Here, an IgA fraction should not be misunderstood as a shortened or degraded IgA. In a preferred embodiment, IgA is full-length IgA including light and heavy chains. IgA as immunoglobulin may also be considered as an antibody (i.e. an IgA antibody).

[0040] IgA derived from body fluids, in particular from plasma, blood plasma or plasma fractions, represents a high number of different antibody specificities. I.e. IgA may be present with different specificities for various antigens, also depending on the choice of body fluid and body fluid donor for obtaining the IgA.

[0041] The body fluid may in principle be any body fluid. In an embodiment of the invention, the body fluid is plasma, blood plasma or a fraction thereof, in particular of a human. Body fluids, in particular plasma, blood plasma or plasma fractions, may be obtained from any source. Body fluids, in particular plasma, blood plasma or plasma fractions, may be obtained from a single donor or may be a pooled fraction of samples. Body fluids, in particular plasma, blood plasma or plasma fractions, may be obtained from naturally immune donors or from immunized or even (hyper)immunized donors. Alternatively or additionally, the body fluid, in particular plasma, blood plasma or plasma fractions, may also contain one or more recombinant components. Alternatively or additionally, the body fluid, in particular plasma, blood plasma or plasma fractions, may also contain one or more monoclonal or polyclonal immunoglobulins, including IgA. The body fluid, in particular plasma, blood plasma or a fraction thereof, may be obtained from a subject (donor) subjected to (hyper)immunization. Alternatively or additionally, selection of the subject (donor) may also involve using commercial vaccines and, optionally, a test system allowing the differentiation between high- and low tittered body fluid, in particular plasma, blood plasma or plasma fractions, donations.

[0042] IgA may be administered by any means. Any route of administration may be applied. For example, IgA may be administered by systemic administration or by local administration. Local administration may be performed by dropping or spraying. Preferably, administration is a local administration, in particular by dropping. In this context, the term "effective amount" as used herein refers to the quantity of IgA needed to achieve the desired therapeutic or preventive outcomes, or to elicit a specific response. The precise effective amount can be determined by those skilled in the art through routine experimentation. For some embodiments of this invention, an effective amount is one that enhances tear secretion in a subject.

[0043] In the context of the application, the term “mainly consist of” is in particular to be understood that occurring impurities consist of the named components by at least 90% (w / w), in particular around 99% (w / w).

[0044] In different embodiments, the said inventive IgA for use is obtained from plasma, blood plasma or plasma fractions at a purity of at least 10% (w / w), at least 25% (w / w), at least 40% (w / w), at least 50% (w / w), at least 60% (w / w), at least 75% (w / w), at least 90% (w / w), at least 95% (w / w), or at least 99% (w / w).

[0045] Impurities consist of proteins typically present in body fluids, in particular in plasma, blood plasma or plasma fractions, such as immunoglobulins other than IgA, the binder and carrier protein albumin, the hydrolase lysozyme, the glycoprotein transferrin, the iron protein complex ferritin, the binder protein fetuin, protease enzymes, or combinations thereof.

[0046] Other immunoglobulins than IgA may be any other types of immunoglobulins (e.g., IgG, IgM, IgD, IgE). In a preferred embodiment, one or more immunoglobulins other than IgA may be selected from the group consisting of immunoglobulin G (IgG), immunoglobulin M (IgM), immunoglobulin D (IgD), immunoglobulin E (IgE), and combinations of two or more thereof. In a preferred embodiment, IgA is separated from IgG and optional further immunoglobulins. In a preferred embodiment, IgA is separated from IgM and optional further immunoglobulins. In a preferred embodiment, IgA is separated from IgG and IgM and optional further immunoglobulins.

[0047] The at least one step of separating IgA from other immunoglobulins may be conducted by any means. In a preferred embodiment, the step of separating IgA from other immunoglobulins may be conducted by means of affinity chromatography.

[0048] In a preferred embodiment, the step of separating IgA from other immunoglobulins may be conducted by means of affinity chromatography wherein one or more agents (affinity ligands) specifically binding to IgA (e.g., one or more IgA-specific antibodies or antibody-fragments) are immobilized on a solid support. Additionally or alternatively, at least one step of separating IgA from other immunoglobulins may involve further procedures such as precipitation (e.g., with ammonium sulfate), size exclusion chromatography, blood plasma fractionation (e.g., Cohn fractionation), ion-exchange (IEX) chromatography, nanofiltration, or a combination of two or more thereof. In a preferred embodiment, at least two or three thereof are combined with each other. Preferably, separating IgA from other immunoglobulins comprises separating IgA from IgG and / or IgM.

[0049] IgA as such may be used in the inventive method. IgA may also be embedded in a pharmaceutical composition, which can have various forms.

[0050] In a preferred embodiment of the inventive immunoglobulin A (IgA) for use said IgA is comprised in a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier, preferably, wherein the pharmaceutical composition is an aqueous solution, in particular wherein the pharmaceutical composition is in the form of eye drops.

[0051] Being comprised in a pharmaceutical composition means, IgA is combined with at least one pharmaceutically acceptable carrier, enabling or facilitating administration. The terms "pharmaceutical composition" and "pharmaceutical formulation" may be understood interchangeably. As used herein, the terms "pharmaceutically acceptable carrier", "pharmaceutically acceptable excipient", "carrier" and "excipient" may be understood interchangeably in the broadest sense as any substance that may support the pharmacological acceptance of the IgA. Such pharmaceutical composition may be ready to use and may preferably be a liquid formulation, in particular an aqueous solution, preferably eye drops. The storage form may also be liquid, but may also be a dried form (e.g. a powder such as a powder comprising dried or freeze-dried IgA) or may be a paste or syrup or the like. Optionally, a dried form, paste or syrup may be dissolved or emulsified prior to being administered to the patient.

[0052] A pharmaceutically acceptable carrier may exemplarily be selected from the list consisting of an aqueous buffer, saline, water, dimethyl sulfoxide (DMSO), ethanol, vegetable oil, paraffin oil or combinations of two or more thereof. Furthermore, the pharmaceutically acceptable carrier may optionally contain one or more detergent(s), one or more foaming agent(s) (e.g., sodium lauryl sulfate (SLS), sodium dodecyl sulfate (SDS)), one or more coloring agent(s) (e.g., food coloring), one or more vitamin(s), one or more salt(s) (e.g., sodium, potassium, calcium, zinc salts), one or more humectant(s) (e.g., sorbitol, glycerol, mannitol, propylenglycol, polydextrose), one or more enzyme(s), one or more preserving agent(s) (e.g., benzoic acid, methylparaben), one or more antioxidant(s), one or more herbal and plant extract(s), one or more stabilizing agent(s), one or more chelating agents (e.g., ethylenediaminetetraacetic acid (EDTA)), and / or one or more uptake mediator(s) (e.g., polyethylene imine (PEI), a cell-penetrating peptide (CPP), a protein transduction domain (PTD), an antimicrobial peptide, etc.).

[0053] For example, the IgA concentration in a pharmaceutical composition may be in the range of 0.01 to 100 mg / mL, or 0.1 to 100 mg / mL, or 0.1 to 10 mg / mL.

[0054] In a preferred embodiment of the inventive immunoglobulin A (IgA) for use, said IgA is obtained from a plasma fraction selected from the group consisting of:

[0055] (a) cryo-poor plasma, typically obtained from the supernatant of blood plasma subjected to being frozen and subsequently thawed;

[0056] (b) a fraction of the Cohn or Kistler-Nitschmann process;

[0057] (c) a plasma fraction obtained from chromatographic methods;

[0058] (d) a plasma fraction obtained from precipitating methods;

[0059] (e) a plasma fraction obtained from filtration methods; or a combination of two or more thereof.

[0060] Plasma, blood plasma or plasma fractions may be obtained from any source. They may be obtained from a single donor or may be a pooled fraction of samples. Such plasma, blood plasma or plasma fraction may be obtained from naturally immune donors or from immunized or even (hyper)immunized donors. Alternatively or additionally, the plasma, blood plasma or plasma fraction may also contain one or more recombinant components. Alternatively or additionally, the plasma, blood plasma or plasma fraction may also contain one or more monoclonal or polyclonal immunoglobulins, including IgA.

[0061] The plasma, blood plasma or plasma fraction may be obtained from a subject (donor) subjected to (hyper)immunization. Alternatively or additionally, selection of the subject (donor) may also involve using commercial vaccines and, optionally, a test system allowing the differentiation between high- and low tittered plasma / blood donations.

[0062] Purification of immunoglobulins by the Cohn process (Cohn et al., Preparation and Properties of Serum and Plasma Proteins. IV. A System for the Separation into Fractions of the Protein and Lipoprotein Components of Biological Tissues and Fluids, 1946, Journal of the American Chemical Society 68 (3): 459-475) and / or Kistler-Nitschmann process (Kistler and Nitschmann, Large Scale Production of Human Plasma Fractions. Eight Years Experience with the Alcohol Fractionation Procedure of Nitschmann, Kistler and Lergier, 1962, Vox Sang 7:414) is known in the art.

[0063] By specific selection of a body fluid donor for obtaining IgA, said IgA may show a bacterial and / or virus specificity. In a preferred embodiment of the inventive immunoglobulin A (IgA) for use, by specific selection of a plasma, blood plasma or plasma fraction donor for obtaining the IgA, said IgA shows a bacterial and / or virus specificity. The “specific selection of a plasma, blood plasma or plasma fraction donor” may be understood as a particular choice of one or more donors as compared to a random selection of donor(s). A subject from which plasma, blood plasma or plasma fraction is obtained may be considered and designated as (plasma, blood plasma or plasma fraction) donor. Plasma, blood plasma or plasma fraction may be obtained from a single donor or may be a pooled fraction of plasma, blood plasma or plasma fraction samples from selected donors.

[0064] Specificities refer to specific antigen targets, in particular bacteria and / or viruses. IgA derived from body fluids, in particular from plasma, blood plasma or plasma fraction, represents a high number of different antibody specificities. By the specific selection of body fluid, in particular plasma, blood plasma or plasma fraction, and / or body fluid, in particular plasma, blood plasma or plasma fraction, donor for obtaining the IgA, preferred specificities for antigens may be selected.

[0065] Therefore, body fluids, in particular plasma, blood plasma or plasma fraction, may be obtained from naturally immune donors or from immunized or even (hyper)immunized donors. Alternatively or additionally, the body fluid, in particular plasma, blood plasma or plasma fraction, from selected donors may also contain one or more recombinant components. Alternatively or additionally, the body fluid, in particular plasma, blood plasma or plasma fraction, from selected donors may also contain one or more monoclonal or polyclonal immunoglobulins, including IgA.

[0066] Thus, the body fluid, in particular plasma, blood plasma or plasma fraction, may be obtained from a subject (donor) subjected to (hyper)immunization. Alternatively or additionally, selection of the subject (donor) may also involve using commercial vaccines and, optionally, a test system allowing the differentiation between highland low tittered body fluid donations.

[0067] Donors may optionally be immunized with a suitable vaccine of choice. Donors may previously be screened for the presence of specific antibodies against specific bacteria and / or viruses, e.g. using ELISA as known in the art. In a preferred embodiment of the inventive immunoglobulin A (IgA) for use, the dry eyes are aqueous production deficient dry eyes and / or evaporative dry eyes. Thus, the dry eyes, in particular marked by tear film instability, result either from insufficient liquid (i.e. tear) production and / or from insufficient liquid (i.e. tear) quality leading to increased evaporation.

[0068] The dry eyes may be caused by various influences such as long-term environmental conditions, conditions of systemic health, short-term expositions, single incidences and / or chronic body conditions.

[0069] In a preferred embodiment of the inventive immunoglobulin A (IgA) for use, the dry eyes are caused by systemic medications, topical treatments, skin conditions around the eyelids, meibomian gland dysfunction, ophthalmic surgery, chemical burns, ocular allergies, decreased androgen levels, excessive or insufficient vitamin intake, prolonged contact lens use, neurotrophic cornea conditions, systemic diseases, environmental factors, lifestyle factors, and / or combinations of two or more thereof.

[0070] In a preferred embodiment of the inventive immunoglobulin A (IgA) for use, the patient has developed or is at risk of developing dry eyes disease (DED), dry eyes syndrome (DES) and / or a chronic ocular, particularly bacterial, viral, or fungal, infection.

[0071] In the context of the application, the term “has developed” as used herein may be understood in the broadest sense in a way that the patient has developed, i.e. is suffering from, a pathological condition associated with disorder associated with dry eyes disease (DED), dry eyes syndrome (DES) and / or a chronic ocular infection. In particular, it may be understood as having a medical diagnosis of dry eyes disease (DED), dry eyes syndrome (DES) and / or a chronic ocular infection.

[0072] The term “at risk of developing” as used herein may particularly be regarded in the context of prophylaxis. It means that the patient has an increased probability of obtaining dry eyes disease (DED), dry eyes syndrome (DES) and / or a chronic ocular infection in comparison to the average probability throughout the population of the same species. More preferably, the risk is at least 5-fold increased, even more preferably the risk is at least 10-fold increased, even more preferably the risk is at least 100-fold increased. The term may also be understood as being exposed to conditions, health issues or the like, promoting a dry eyes disease (DED), dry eyes syndrome (DES) and / or a chronic ocular infection, the (increased) risk preferably being medically diagnosed. In a preferred embodiment of the inventive immunoglobulin A (IgA) for use, the patient is characterized in that the IgA level in the lacrimal fluid of the eyes of the patient is decreased by at least 15% in comparison to the average level found throughout the population of the same species.

[0073] In another embodiment, the IgA level in the lacrimal fluid of the eyes is decreased by at least 20%, or by at least 25%, or by at least 30%, or by at least 40%, or by at least 50%, in comparison to the average level found throughout the population of the same species. Administration may be adjusted to the IgA levels analyzed in the lacrimal fluid.

[0074] In a preferred embodiment of the inventive immunoglobulin A (IgA) for use, the patient has developed or is at risk of developing acute symptoms referring to dry eyes comprising stinging, burning, irritation, dryness, redness, gritty sensation in the eye, excessive tearing, blurred vision and / or combinations thereof.

[0075] The patient may be administered with IgA according to any administration scheme. In this context, a single dose may be a single dose in any dosage range. In a preferred embodiment, administration of a single dose may be each in the range of 0.01 to 100 mg. Preferably, the patient is administered via IgA containing eye drops. The patient may be administered with IgA at least once per day, at least once every other day, or at least once per week for a period of at least one day, at least three days, at least one week, three to seven days at least two weeks, at least four weeks, at least two months, or at least a year. For example, administration may be once every two weeks, once every week or twice daily (e.g. for prophylaxis). It may also be once, twice, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times or 10 times or even every hour or bihourly.

[0076] The patient may be a healthy patient or a patient suffering from dry eyes. In particular, the patient has developed dry eyes disease (DED), dry eyes syndrome (DES) and / or a chronic ocular, particularly bacterial, viral, or fungal, infection.

[0077] Eye drops may contain IgA e.g. at a concentration from 0.01 mg / mL to 500 mg / mL, or from 0.01 mg / mL to 100 mg / mL, or from 0.05 mg / mL to 100 mg / mL, or from 0.1 mg / mL to 100 mg / mL, or from 0.1 mg / mL to 10 mg / mL. In one embodiment, eye drops may contain IgA at a concentration from 0.05 mg / mL to 1 mg / mL, or from 0.08 mg / mL to 0.5 mg / mL.

[0078] In one embodiment, eye drops may also contain IgG and / or IgM at a concentration from 0.05 mg / mL to 1 mg / mL, or from 0.08 mg / mL to 0.5 mg / mL. In a preferred embodiment of the inventive immunoglobulin A (IgA) for use, the patient’s eye is administered with IgA containing eye drops at a concentration from 0.05 to 100 mg / mL at least once with a dose of IgA in the range of 0.01 to 10 mg; at least once within one week before being subjected to an event of high risk of developing dry eyes such as a surgical intervention with a dose of IgA in the range of 0.01 to 10 mg; or, on a regular basis when being at risk of developing dry eyes with a dose of IgA in the range of 0.01 to 10 mg; and / or according to one of the following administration schemes:

[0079] (A) administration of IgA at least once per day for a time period of three or more days of a single dose each in the range of 0.01 to 10 mg;

[0080] (B) administration of IgA at least three times a day for a time period of three or more days of a single dose each in the range of 0.01 to 10 mg; or

[0081] (C) hourly, bihourly, several times per day according to the subjective perception.

[0082] Eye drops are typically administered at drop sizes from 10 pL to 100 pL, but may also have larger volumes. Preferably, drop sizes are 10-100 pL.

[0083] The patient may be administered according to an administration scheme as outlined above for a time period of three or more days, preferably for a time period of at least a week, for at least two weeks, at least four weeks, at least two months, or at least a year.

[0084] The present invention also relates to a pharmaceutical composition comprising a protein composition, wherein said protein composition comprises 0.01 -100% (w / w), in particular 0.01 -50% (w / w), preferably at least 0.1 % (w / w), more preferably at least 1 % (w / w), even more preferably at least 10% (w / w), based on the total polypeptide mass, of Immunoglobulin A (IgA) obtained from plasma, blood plasma or plasma fractions.

[0085] The protein composition, comprised in the pharmaceutical composition, preferably comprises at least 0.01 % (w / w), at least 0.1 % (w / w), at least 1 % (w / w), at least 5% (w / w), at least 10% (w / w), at least 15% (w / w), at least 20% (w / w), at least 25% (w / w), at least 30% (w / w), at least 40% (w / w), at least 50% (w / w), at least 60% (w / w), at least 75% (w / w), at least 80% (w / w), at least 85% (w / w), at least 90% (w / w), at least 95% (w / w), at least 98% (w / w), at least 99% (w / w), based on the total polypeptide mass, of IgA obtained from plasma, blood plasma or plasma fractions.

[0086] The protein composition, comprised in the pharmaceutical composition may e.g. comprise 0.01 -100% (w / w), 0.01 -99% (w / w), 0.01 -90% (w / w), 0.01 -50% (w / w), 0.1-50% (w / w), 1 -50% (w / w), 5-50% (w / w), 10-50% (w / w), 10-60% (w / w), 20-70% (w / w), 30-80% (w / w), based on the total polypeptide mass, of IgA obtained from a body fluid.

[0087] The pharmaceutical composition, in particular in form of eye drops, differs from naturally occurring lacrimal fluid in the comprised protein components and their ratios. IgA may be in concentrations as in natural lacrimal fluid (see Table 1 ) or may be enriched. Other components, such as proteins naturally occurring (see Table 1 ), may not be present or only marginal. Preferably, IgA is the only main component of the inventive pharmaceutical composition, in particular the eye drops.

[0088] One embodiment also relates to a dosage unit of the pharmaceutical composition usable in the context of the treatment or prevention as outlined in the context of the invention. Exemplarily, one embodiment may refer to a single dose container or to a multiple dosage form. The dose range may be adjusted to the intended use.

[0089] In a preferred embodiment, the inventive pharmaceutical composition comprises a bacterial and / or virus specific IgA, created by specific selection of a body fluid, in particular plasma, blood plasma or plasma fraction, donor for obtaining the IgA.

[0090] The “specific selection of a body fluid, in particular plasma, blood plasma or plasma fraction, donor” may be understood as a particular choice of one or more donors as compared to a random selection of donor(s). A subject from which body fluid, in particular plasma, blood plasma or plasma fractions, is obtained may be considered and designated as (body fluid, in particular plasma, blood plasma or plasma fraction) donor. Body fluids, in particular plasma, blood plasma or plasma fractions, may be obtained from a single donor or may be a pooled fraction of body fluid, in particular plasma, blood plasma or plasma fractions, samples from selected donors. Specificities refer to specific antigen targets, in particular bacteria and / or viruses. IgA derived from body fluids, in particular plasma, blood plasma or plasma fractions, represents a high number of different antibody specificities.

[0091] By the specific selection of body fluid, in particular plasma, blood plasma or plasma fractions, and / or body fluid, in particular plasma, blood plasma or plasma fraction, donor for obtaining the IgA, preferred specificities for antigens may be selected. Therefore, body fluids, in particular plasma, blood plasma or plasma fractions, may be obtained from naturally immune donors or from immunized or even (hyper)immunized donors. Alternatively or additionally, the body fluid, in particular plasma, blood plasma or plasma fractions, from selected donors may also contain one or more recombinant components. Alternatively or additionally, the body fluid, in particular plasma, blood plasma or plasma fractions, from selected donors may also contain one or more monoclonal or polyclonal immunoglobulins, including IgA.

[0092] Thus, the body fluid, in particular plasma, blood plasma or plasma fractions, may be obtained from a subject (donor) subjected to (hyper)immunization. Alternatively or additionally, selection of the subject (donor) may also involve using commercial vaccines and, optionally, a test system allowing the differentiation between high- and low tittered body fluid donations. Donors may optionally be immunized with a suitable vaccine of choice. Donors may previously be screened for the presence of specific antibodies against specific bacteria and / or viruses, e.g. using ELISA as known in the art.

[0093] In a preferred embodiment of the invention, the pharmaceutical composition is for use in a method for treating or preventing dry eyes in a patient. In a preferred embodiment of the invention, the pharmaceutical composition is for use in a method for treating or preventing infections caused by dry eyes in a patient. In a preferred embodiment of the invention, the pharmaceutical composition is for use in a method for treating or preventing bacterial, viral, or fungal ocular infections of a patient. The inventive pharmaceutical composition may be for use in a method for treating or preventing dry eyes in a patient, and / or infections caused by dry eyes in a patient, and / or bacterial, viral, or fungal ocular infections of a patient.

[0094] Further disclosed is Immunoglobulin A (IgA) obtained from body fluids at a purity of at least 10% (w / w), preferably at least 25% (w / w), more preferably at least 40% (w / w), even more preferably at least 50% (w / w), in particular wherein impurities mainly consist of immunoglobulins other than IgA, in particular IgG and / or IgM, albumin, lysozyme, transferrin, ferritin, fetuin and / or proteases.

[0095] The invention further relates to Immunoglobulin A (IgA) obtained from plasma, blood plasma or plasma fractions at a purity of at least 10% (w / w), preferably at least 25% (w / w), more preferably at least 40% (w / w), even more preferably at least 50% (w / w), in particular wherein impurities mainly consist of immunoglobulins other than IgA, in particular IgG and / or IgM, albumin, lysozyme, transferrin, ferritin, fetuin and / or proteases. Herein, “mainly consist of” means that occurring impurities consist of the named components by at least 90% (w / w), in particular around 99% (w / w).

[0096] In different embodiments, the IgA obtained from plasma, blood plasma or plasma fractions is of purity of at least 10% (w / w), at least 25% (w / w), at least 40% (w / w), at least 50% (w / w), at least 60% (w / w), at least 75% (w / w), at least 90% (w / w), at least 95% (w / w), or at least 99% (w / w).

[0097] Impurities consist of proteins typically present in body fluids, in particular in plasma, blood plasma or plasma fractions, such as immunoglobulins other than IgA, the binder and carrier protein albumin, the hydrolase lysozyme, the glycoprotein transferrin, the iron protein complex ferritin, the binder protein fetuin, protease enzymes, or combinations thereof.

[0098] Other immunoglobulins than IgA may be any other types of immunoglobulins (e.g., IgG, IgM, IgD, IgE). In a preferred embodiment, one or more immunoglobulins other than IgA may be selected from the group consisting of immunoglobulin G (IgG), immunoglobulin M (IgM), immunoglobulin D (IgD), immunoglobulin E (IgE), and combinations of two or more thereof. In a preferred embodiment, IgA is separated from IgG and optional further immunoglobulins. In a preferred embodiment, IgA is separated from IgM and optional further immunoglobulins. In a preferred embodiment, IgA is separated from IgG and IgM and optional further immunoglobulins.

[0099] The at least one step of separating IgA from other immunoglobulins may be conducted by any means. In a preferred embodiment, the step of separating IgA from other immunoglobulins may be conducted by means of affinity chromatography. In a preferred embodiment, the step of separating IgA from other immunoglobulins may be conducted by means of affinity chromatography wherein one or more agents (affinity ligands) specifically binding to IgA (e.g., one or more IgA-specific antibodies or antibody-fragments) are immobilized on a solid support.

[0100] Additionally or alternatively, at least one step of separating IgA from other immunoglobulins may involve further procedures such as precipitation (e.g., with ammonium sulfate), size exclusion chromatography, blood plasma fractionation (e.g., Cohn fractionation), ion-exchange (IEX) chromatography, nanofiltration, or a combination of two or more thereof.

[0101] In a preferred embodiment, at least two or three thereof are combined with each other. Preferably, separating IgA from other immunoglobulins comprises separating IgA from IgG and / or IgM. The invention relates to Immunoglobulin A (IgA) obtained from body fluids, in particular from plasma, blood plasma or plasma fractions, at a purity of at least 50%, wherein impurities mainly consist of immunoglobulins other than IgA, albumin, lysozyme, transferrin, ferritin, fetuin and / or proteases.

[0102] In a preferred embodiment, the Immunoglobulin A (IgA) is for use according to the invention or in an inventive pharmaceutical composition, as described within this application.

[0103] Preparation of IgA can be achieved by the following process:

[0104] Purification of the immunoglobulins by the known Cohn- [Cohn, E. J.; Strong, L. E.; Hughes, W. L.; Mulford, D. J.; Ashworth, J. N.; Melin, M.; Taylor, H. L. (1946). "Preparation and Properties of Serum and Plasma Proteins. IV. A System for the Separation into Fractions of the Protein and Lipoprotein Components of Biological Tissues and Fluids1 a,b,c,d". Journal of the American Chemical Society. 68 (3): 459- 475] or Kistler-Nitzschman-fractionation [P. Kistler, HS. Nitschmann: Large Scale Production of Human Plasma Fractions. Eight Years Experience with the Alcohol Fractionation Procedure of Nitschmann, Kistler and Lergier. VoxSang 7 1962, 414],

[0105] The separation of IgA and IgG can be done either ahead of the positive affinity chromatography or after this step by a second affinity chromatography step using an affinity ligand, like CaptureSelect™ IgA or CaptureSelect™ lgA-CH1 by Thermo-fisher or any other IgA-specific ligand being either a whole protein or just a peptide fragment, like a fab fragment. Another option for the purification of IgA lies in the separation of the IgM and IgA waste fraction, which is generated during the polishing of IgG products in Cohn Fractionation. The IgA in this fraction can be separated using IEX Chromatography and or nanofiltration to remove IgM and purify IgA.

[0106] The present invention is further illustrated by the following examples, figures and claims.

[0107] Brief Description of the Figures

[0108] Figure 1 Tear-volume evaluation in DED animal model and controls before and after IgA treatment; *** p<0.11 .; ** p<0.01 ; *p<0.05;

[0109] Figure 2 Inflammatory Index Evaluation in DED animal model and controls before and after IgA treatment; *** p<0.11 .; ** p<0.01 ; *p<0.05;

[0110] Figure 3 Tear Break up Time in DED animal model and controls before and after IgA treatment; *** p<0.11.; ** p<0.01 ; *p<0.05; Figure 4 Ocular surface staining using 0.1 % Sodium fluorescein of healthy eye and eyes in DED animal model and controls before and after IgA treatment. 1 ) Control shows clear iris and pupil details; 2) after 7 days DED model shows wrinkled and no corneal architecture; on day 13: 3) after treatment with IgA [2.5pg], there are notable ciliary hyperaemia and oedema, the whole eye anatomy has not restored; 4) after treatment with IgA [5pg] there is no dryness, inflammation with slight corneal oedema; 5) in the prophylaxis group almost near normal appearance is found; 6) after treatment with albumin, eye anatomy has not completely restored, with slight leathery texture;

[0111] Figure 5 Inflammatory Index Evaluation in DED animal model, DED and bacterial infection model, and controls after IgA treatment.

[0112] Examples

[0113] Obtaining IgA from body fluids, in particular from plasma, blood plasma or plasma fractions, may be conducted as known from EP-A 3907238.

[0114] Example I: Purification of IgA by Crohn fractionation and affinity chromatography

[0115] An IgA containing fraction is isolated directly from plasma or from plasma fractions, which are purified from IgG. When those fractions are used, the separation of the specific immunoglobulin classes is not needed. When the positive affinity chromatography is performed ahead of the separation of the immunoglobulin classes, the separation of them is performed in a second step.

[0116] An example for the separation of (hyperimmune) IgA includes subjecting a plasma pool to Cohn fractionation. This is followed by an IgG polishing step providing IgG and an IgA / IgM fraction. The IgA / IgM fraction is subjected to affinity chromatography and provides IgA.

[0117] An alternative example for the separation of IgA includes subjecting an un-fractioned plasma pool to affinity chromatography. This provides an immunoglobulin fraction (including IgG and IgA) and a residual blood fraction, which is further subjected to Cohn fractionation. The immunoglobulin fraction (including IgG and IgA) is subjected to a second affinity chromatography separating IgG and IgA and provides IgA.

[0118] Example II: IgA purification by chromatography

[0119] The feed for performing chromatography can consist e.g. either of plasma, cryopoor plasma or of any plasma derived solution which is produced during plasma fractionation. The feed is diluted in equal parts with 1x PBS buffer. The resin (CaptureSelect IgA) is equilibrated with 1x PBS buffer and is used either in a chromatography column or in batch-mode. The solution is loaded onto the chromatography column or incubated with the resin in batch-mode. The elution is performed with a 0.1 M glycine, pH 3 or lower. 1 M citric acid is then used to CIP the column, which is then equilibrated afterwards using 1x PBS again.

[0120] Example III: Animal model of dry eye syndrome

[0121] Induction of the dry eye syndrome Benzalkonium chloride (BAC) is a widely used preservative in ophthalmic solutions; however, it has been identified as a potential contributor to dry eye syndrome. Consequently, numerous in vitro and in vivo models of dry eye syndrome have been effectively developed using BAC.

[0122] Based on the experimental evidence provided by these models, the drug molecule Immunoglobulin A (IgA) has been found to be an effective ophthalmic drug to treat or prevent dry eye disease / syndrome.

[0123] Comparison with other compositions surprisingly had shown that treatment with an IgM containing product or an albumin containing product had no effect, while an IgG containing product even worsened the dry eyes of the patient (no data shown in application).

[0124] Benzalkonium chloride induced dry eye mice model [Xiao.et.al 2012]

[0125] Male BALB / c mice (6-8 weeks) were used for this study.

[0126] These mice were kept in the facility with standard environment throughout the study as follows: room temperature 25 ± 1 °C, relative humidity 60% ± 10%, and alternating 12-hour light-dark cycles.

[0127] For the study animals were taken into 6 groups comprising a control group, a disease control [DED] (untreated), a disease group treated with drug molecule IgA at concentration of 2.5 pg / dose, a disease group treated with drug molecule IgA at concentration of 5 pg / dose, a prophylaxis group treated with drug molecule IgA (5 pg / dose) at an early stage and a disease group treated with Albumin (5 pg / dose).

[0128] The groups’ details are summarized in Table-2. Table 2. Summary of groups included in the study. Abbreviations: IgA - Immunoglobulin A; DED - Dry Eye Disease; BAC - Benzalkonium chloride; TID - three times a day

[0129] The following parameters used in the study to evaluate the DED induction and efficacy of IgA

[0130] Tear volume evaluation

[0131] Tear volume was measured by the phenol red thread tear test using cotton threads in the standard environment. Mice were kept immobile by intraperitoneal injection of 60 mg / kg pentobarbital. The lower eyelid was pulled down slightly, and a 1 mm portion of the thread was placed on the palpebral conjunctiva for 15 seconds at a specified point approximately one third of the distance from the lateral canthus of the lower eyelid. The red portion of the thread was measured in millimeters.

[0132] Tear Break-Up Time (TBUT) and Fluorescein Staining

[0133] One microliter of 0.1 % liquid sodium fluorescein was dropped into the conjunctival sac. After 3 blinks TBUT was recorded in seconds. Ninety seconds later, corneal epithelial damage was graded with a cobalt blue filter under a slit-lamp microscope. The cornea was divided into four quadrants, which was scored respectively. The four scores were added to a final grade (total, up to 16 points). The fluorescein score was analyzed as previously described with essential modification; as follows: absent, 0; slightly punctuate staining <30 spots, 1 ; punctate staining >30 spots, but not diffuse, 2; severe diffuse staining but no positive plaque, 3; and positive fluorescein plaque, 4.

[0134] Evaluation of Inflammation index

[0135] The inflammatory index was evaluated, based on three parameters: ciliary hyperemia (absent, 0; present but <1 mm, 1 ; present between 1 and 2 mm, 2; present >2 mm, 3); central corneal edema (absent, 0; present with visible iris details, 1 ; present without visible iris details, 2; present without visible pupil, 3); and peripheral corneal edema (absent, 0; present with visible iris details, 1 ; present without visible iris details, 2; present with no visible iris, 3). The final inflammatory index result was obtained by summing the scores of the different parameters and then dividing by a factor of nine.

[0136] Histopathological Evaluation by H&E staining

[0137] The following Corneal and conjunctival pathological changes were observed in Corneal histopathological findings:

[0138] * Epithelial changes: Thinning, irregularity, microcysts, loss of microvilli

[0139] * Stromal changes: Demarcation lines (Khodadoost lines), hyper reflectivity, nerve fiber loss

[0140] * Sub epithelial haze: Deposition of inflammatory cells and debris

[0141] * Goblet cell loss: Reduced tear production

[0142] * Corneal erosion: Desquamation of the epithelium due to dryness

[0143] Conjunctival histopathological findings:

[0144] * Epithelial changes: Hyperplasia, goblet cell loss, squamous metaplasia

[0145] * Sub epithelial fibrosis: Collagen deposition in the stroma

[0146] * Lymphoid follicles: Aggregation of immune cells

[0147] * Vascularization: Increased blood vessel formation

[0148] Grades:

[0149] * Grade 0: Normal conjunctiva with no goblet cell loss or squamous metaplasia;

[0150] * Grade 1: Mild changes, including slight goblet cell loss and minimal squamous metaplasia / hyperplasia / erosion / flattening / cyst / vacuolation of epithelial cells / Sub epithelial inflammation / fibrosis involving up to 25% of the surface area; * Grade 2: Moderate changes, including slight goblet cell loss and minimal squamous metaplasia / hyperplasia / erosion / flattening / cyst / vacuolation of epithelial cells / Sub epithelial inflammation / fibrosis involving up to 50% of the surface area;

[0151] * Grade 3: Severe changes, including slight goblet cell loss and minimal squamous metaplasia / hyperplasia / erosion / flattening / cyst / vacuolation of epithelial cells / Sub epithelial inflammation / fibrosis involving up to 75% of the surface area.

[0152] PAS Staining Scoring or Grading System

[0153] Epithelial cells of cornea / conjunctiva and goblet cells in conjunctival epithelial cells secrete mucin stain positive for PAS with pink color. The level of presence of mucin is graded as follows:

[0154] 0- Normal level of mucin in epithelial cells of cornea / conjunctiva and goblet cells in conjunctival epithelial cells;

[0155] 1 - Mild increase level of mucin in epithelial cells of cornea / conjunctiva and goblet cells in conjunctival epithelial cells;

[0156] 2- Moderate increase level of mucin in epithelial cells of cornea / conjunctiva and goblet cells in conjunctival epithelial cells;

[0157] 3- Severe increase level of mucin in epithelial cells of cornea / conjunctiva and goblet cells in conjunctival epithelial cells.

[0158] Experimental outline and conduction

[0159] Before the initiation of disease induction, parameters such as tear volume, tear breakup time and inflammatory index were recorded.

[0160] Both eyes of these mice were topically administered twice daily with 5 pL of 0.1 % Benzalkonium Chloride (BAC) for 6 days. The parameters such as tear volume, tear breakup time and inflammatory index were recorded at Day 7 and treatment was initiated.

[0161] Except the control and disease control group, the other groups were subjected to treatment with 5 pL of the drug molecule IgA at concentrations of 2.5 pg / dose and 5 pg / dose for respective groups for 12 days. On Day 13, the parameters were measured and results are shown in Fig.1 , Fig.2, Fig.3. The H&E and PAS staining were graded according to Nelson’s grading system and evaluated statistically. Data not shown.

[0162] Unpaired two tailed t-test was used for analysis of the data and *** p<0.001 ; ** p<0.01 ;

[0163] *p<0.05 compared to both the control and the disease control group. The mice treated with 2.5 pg / dose showed notable ciliary hyperaemia and oedema, the whole eye anatomy was not restored. The mice treated with 5 pg / dose showed the restoration of complete eye anatomy that indicated a potential of the IgA to reverse the dry eye condition. The mice of the prophylaxis group also showed restoration of corneal architecture, which suggests the usage of IgA as a prophylaxis treatment in the early stages of the disease.

[0164] Example IV: Animal model of dry eye syndrome and infected dry eyes

[0165] Induction of dry eye syndrome and bacterial infection: Dry eye syndrome was induced as described above. For bacterial infection, both eyes of the animals were inoculated with 0.5 cfu / eye (5pL) of 100 cfu / mL Staphylococcus aureus through topical application.

[0166] Mice were selected and kept as outlined above. For the study animals were taken into 9 groups comprising a control group, a disease control [DED] (untreated), a disease group treated with drug molecule IgA at concentration of 2.5 pg / dose, a disease group treated with drug molecule IgA and Albumin both at a concentration of 2.5 pg / dose, a disease group treated with Albumin (2.5 pg / dose), a bacterial infection control (untreated), a DED and bacterial infection control (untreated), a disease and bacterial infection group treated with drug molecule IgA at concentration of 2.5 pg / dose at an early stage (prophylaxis group) and a disease and bacterial infection group treated with drug molecule IgA at concentration of 2.5 pg / dose (treatment group).

[0167] The groups’ details are summarized in Table-3.

[0168] Table 3. Summary of groups included in the study. Abbreviations: IgA - Immunoglobulin A; DED - Dry Eye Disease; BAC - Benzalkonium chloride; cfu - colony forming units; TID - three times a day

[0169] The parameters used in the study to evaluate the DED induction and efficacy of IgA were as described in Example III.

[0170] Experimental outline and conduction

[0171] The objective of this study was to assess the effect of IgA Eye drops on Balb / c mice by comparing the animals subjected to Dry Eye Disease induced by Benzalkonium

[0172] Chloride and DED with infection by Staphylococcus aureus. In this experiment, a total of 45 male Balb / c mice were divided into 9 groups (G1 to G9). All the eyes of the animals except Groups 1 and 6 were instilled with 0.1 % BAC twice a day for 7 days and paused for 2 days due to severity. Groups 1 and 6 did not receive BAC and were considered as control. G2 received 0.1 % BAC (5 pL) without any treatment served as disease control. G3 - G5 were subjected to 0.1 % BAC and were treated with IgA, IgA + Albumin, and Albumin respectively. The treatment dose was applied topically to both the eyes at 2.5 pg / dose. Both eyes of mice from G6 to G9 were subjected to bacterial infection at 100 cfu / mL (5 pL was instilled into the eye to infect). Animals from G6 served as control group and were subjected to bacterial infection only. G7 - G9 groups were subjected to DED and bacterial infection at 100 cfu / mL.

[0173] Animals from G7 were subjected to DED and bacterial infection without a treatment. Animals from G8, were subjected to DED, prophylactic treatment with IgA three days prior to bacterial infection.

[0174] Animals from G9 were subjected to DED, bacterial infection and treatment with IgA. Animals were treated for 10 days after the induction of DED for G1 -G5 groups and for G6-G9 after bacterial infection except for prophylactic treatment group that was treated for 13 days (3 days prior and 10 days after bacterial infection including the day of infection). The inflammatory index was despite of the infection reduced as shown in Fig.5.

[0175] No synergistic effect was found in the treatment group G4 (IgA + Albumin). Prophylaxis group was able to regain the architecture of the eye partially, and the bacterial infection was interrupted by day 10 after treatment.

[0176] The animals treated with IgA only (G9), was observed to have regained the complete architecture of the eye if the treatment was extended.

Claims

PreviPharma GmbH October 30, 2025Claims1 . Immunoglobulin A (IgA) for use in a method for treating or preventing dry eyes in a patient wherein said IgA is obtained from plasma, blood plasma or plasma fractions at a purity of at least 50%, wherein impurities mainly consist of immunoglobulins other than IgA, albumin, lysozyme, transferrin, ferritin, fetuin and / or proteases, and wherein said IgA is administered to said patient in an effective dosage.

2. The Immunoglobulin A (IgA) for use according to claim 1 wherein said IgA is comprised in a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier, preferably, wherein the pharmaceutical composition is an aqueous solution, in particular wherein the pharmaceutical composition is in the form of eye drops.

3. The Immunoglobulin A (IgA) for use according to claim 1 or 2 wherein said IgA is obtained from a plasma fraction selected from the group consisting of:(a) cryo-poor plasma, typically obtained from the supernatant of blood plasma subjected to being frozen and subsequently thawed;(b) a fraction of the Cohn or Kistler-Nitschmann process;(c) a plasma fraction obtained from chromatographic methods;(d) a plasma fraction obtained from precipitating methods;(e) a plasma fraction obtained from filtration methods; or a combination of two or more thereof.

4. The Immunoglobulin A (IgA) for use according to any of claims 1 -3 wherein by specific selection of a plasma, blood plasma or plasma fraction donor for obtaining the IgA, said IgA shows a bacterial and / or virus specificity.

5. The Immunoglobulin A (IgA) for use according to any of claims 1 -4 wherein the dry eyes are aqueous production deficient dry eyes and / or evaporative dry eyes.

6. The Immunoglobulin A (IgA) for use according to any of claims 1 -5 wherein the dry eyes are caused by systemic medications, topical treatments, skin conditions around the eyelids, meibomian gland dysfunction, ophthalmic surgery, chemical burns, ocular allergies, decreased androgen levels, excessive or insufficient vitamin intake, prolonged contact lens use,neurotrophic cornea conditions, systemic diseases, environmental factors, lifestyle factors, and / or combinations of two or more thereof.

7. The Immunoglobulin A (IgA) for use according to any of claims 1 -6 wherein the patient has developed or is at risk of developing dry eyes disease (DED), dry eyes syndrome (DES) and / or a chronic ocular bacterial, viral, or fungal infection.

8. The Immunoglobulin A (IgA) for use according to any of claims 1 -7 wherein the patient is characterized in that the IgA level in the lacrimal fluid of the eyes is decreased by at least 15% in comparison to the average level found throughout the population of the same species.

9. The Immunoglobulin A (IgA) for use according to any of claims 1 -8 wherein the patient has developed or is at risk of developing acute symptoms referring to dry eyes comprising stinging, burning, irritation, dryness, redness, gritty sensation in the eye, excessive tearing, blurred vision and / or combinations thereof.

10. The Immunoglobulin A (IgA) for use according to any of claims 1 -9 wherein the patient’s eye is administered with IgA containing eye drops at a concentration from 0.1 to 100 mg / mL; at least once with a dose of IgA in the range of 0.01 to 10 mg; at least once within one week before being subjected to an event of high risk of developing dry eyes such as a surgical intervention with a dose of IgA in the range of 0.01 to 10 mg; or on a regular basis when being at risk of developing dry eyes with a dose of IgA in the range of 0.01 to 10 mg; and / or according to one of the following administration schemes:(A) administration of IgA at least once per day for a time period of three or more days of a single dose each in the range of 0.01 to 10 mg;(B) administration of IgA at least three times a day for a time period of three or more days of a single dose each in the range of 0.01 to 10 mg; or(C) hourly, bihourly, several times per day according to the subjective perception.

11. A pharmaceutical composition comprising a protein composition, wherein said protein composition comprises 0.01-50% (w / w), preferably at least 0.1 % (w / w), based on the total polypeptide mass, of Immunoglobulin A (IgA) obtained from plasma, blood plasma or plasma fractions.

12. The pharmaceutical composition of claim 11 , wherein a bacterial and / or virus specific IgA is comprised, created by specific selection of a plasma, blood plasma or plasma fraction donor for obtaining the IgA.

13. The pharmaceutical composition of claim 11 or 12 for use in a method for treating or preventing dry eyes in a patient, and / or infections caused by dry eyes in a patient, and / or bacterial, viral, or fungal ocular infections of a patient.

14. Immunoglobulin A (IgA) obtained from plasma, blood plasma or plasma fractions, at a purity of at least 50%, wherein impurities mainly consist of immunoglobulins other than IgA, albumin, lysozyme, transferrin, ferritin, fetuin and / or proteases.

15. Immunoglobulin A (IgA) according to claim 14 for use in a method for treating or preventing dry eyes in a patient or for use in a pharmaceutical composition.

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