Il-17c based therapeutics for promoting epithelial healing and inhibiting fibrosis in ocular disorders

Abstract

Disclosed are pharmaceutical compositions for ocular administration comprising interleukin-17C (IL-17C) as protein, messenger RNA encoding IL-17C, or IL-17C protein encapsulated in a delivery vehicle. The compositions include delivery systems selected from lipid nanoparticles (LNPs), exosomes, hydrogels, and ocular inserts. In certain embodiments, the LNP formulation comprises an ionizable lipid, cholesterol, a phospholipid, and a PEG-lipid, with particles sized for ocular use; in other embodiments, exosomes derived from mesenchymal stem cells, corneal epithelial cells, or dendritic cells are loaded with IL-17C protein or IL-17C mRNA. IL-17C mRNA embodiments may include modified nucleotides and 5' cap / UTR / poly(A) features. When evaluated in relevant models, the compositions reduce alpha-SMA and collagen gel contraction, and decrease corneal opacity and neovascularization while enhancing epithelial wound closure and nerve metrics. Also disclosed is a sustained-release ocular device comprising the compositions, including hydrogel implants, bandage contact lenses, or biodegradable ocular inserts.

Description

IL-17C BASED THERAPEUTICS FOR PROMOTING EPITHELIAL HEALING AND INHIBITING FIBROSIS IN OCULAR DISORDERSFIELD OF INVENTION

[0001] The present invention relates to therapeutic treatments for ocular surface disorders. In particular, the present invention concerns the use of interleukin- 17C (IL-17C) in promoting epithelial healing, regulating cellular proliferation, and inhibiting fibrosis in ocular tissues.BACKGROUND

[0002] Ocular surface disorders encompass a broad range of diseases and conditions affecting the cornea, conjunctiva, and other structures that play critical roles in maintaining healthy vision. These disorders can result from various factors, including trauma, surgery, infections, immune -related inflammation, or chronic conditions such as dry eye disease, Stevens-Johnson syndrome, vernal keratoconjunctivitis, and keratoconus, among others. Common among these conditions is a disruption of the delicate balance between wound healing and tissue regeneration, which is essential for maintaining the integrity of the ocular surface.

[0003] A primary challenge in managing ocular surface disorders is the impairment of epithelial healing. The corneal epithelium, the outermost layer of the eye, is particularly susceptible to injury from both external and internal factors. When epithelial damage occurs, the body initiates a complex wound -healing response involving cellular migration, proliferation, and differentiation. However, in many cases, this process is disrupted, leading to delayed re-epithelialization or chronic wounds. Impaired epithelial healing can cause pain, vision impairment, and increased risk of infection, often resulting in further complications. One significant issue in the current management of these disorders is the development of fibrosis during or after the healing process. Fibrosis, characterized by excessive extracellular matrix deposition, particularly collagen and fibronectin, leads to scarformation and opacity in the cornea, which can severely compromise vision. Transforming Growth Factor-Beta (TGF-P) is a key mediator in this fibrotic response. Under normal conditions, TGF-P plays a role in wound healing, but excessive or prolonged activation of TGF-P signaling can result in uncontrolled fibrotic tissue formation. This is a major complication in post-surgical recovery, such as following corneal transplantation, or in chronic inflammatory conditions like dry eye syndrome and ocular cicatricial pemphigoid.

[0004] Current treatments for ocular surface disorders include topical corticosteroids, anti-inflammatory agents, and growth factor-based therapies, all of which aim to modulate inflammation and promote healing. However, these treatments often come with significant limitations. Long-term use of corticosteroids, for example, can cause side effects such as increased intraocular pressure, glaucoma, or cataract formation. Furthermore, while some treatments may reduce inflammation, they do not adequately address the fibrotic component of the healing process, leading to suboptimal recovery outcomes. The persistence of fibrosis and scarring after injury or surgery remains a significant unmet clinical need in ophthalmology. Additionally, many available therapies fail to target the underlying cellular and molecular mechanisms involved in epithelial wound healing and fibrosis. In particular, the role of specific cytokines and their impact on epithelial cell behavior and immune modulation is not fully understood or exploited in current therapeutic strategies.

[0005] Thus, given the complex and multifactorial nature of ocular surface disorders, there remains a critical need for more effective therapeutic strategies that not only promote epithelial healing but also prevent the progression of fibrosis and scar tissue formation, while minimizing adverse effects.SUMMARY OF THE INVENTION

[0006] The present invention discloses a therapeutic method for managing and treating various ocular surface disorders by utilizing the unique biological properties of interleukin- 17C (IL-17C). Ocular surface disorders, including cornealulcers, dry eye syndrome, and other inflammatory conditions, often lead to impaired epithelial healing and the development of fibrosis, which can result in significant vision impairment and reduced quality of life. Current treatment options, such as anti-inflammatory drugs and corticosteroids, may address inflammation but often fall short in promoting effective epithelial regeneration or preventing fibrosis, leading to suboptimal therapeutic outcomes.

[0007] In an embodiment, the present invention specifically targets the dual role of IL-17C in enhancing epithelial wound healing while concurrently inhibiting fibrosis, which represents a significant improvement over current therapies. IL-17C, a member of the IL- 17 cytokine family, plays a critical role in epithelial immunity and tissue repair. It is predominantly expressed by epithelial cells and acts through its receptor, IL-17RE, which is also found in epithelial cells. The autocrine signaling mechanism of IL-17C promotes epithelial cell proliferation, re-epithelialization, and recovery from injury, making it a crucial factor in maintaining ocular surface integrity.

[0008] In addition to its regenerative properties, IL-17C regulates the activity of Transforming Growth Factor-Beta (TGF-B), a key driver of fibrosis. Excessive TGF-B signaling leads to uncontrolled extracellular matrix deposition, collagen formation, and tissue scarring, particularly in the cornea, where fibrosis can severely impact visual function. The present invention exploits IL-17C's ability to counteract these fibrotic processes by inhibiting TGF-B-driven pathways, thereby preventing or reducing the formation of fibrotic tissue, collagen, and fibronectin deposition during wound healing. The therapeutic compositions provided in the present invention consist of IL-17C-based agents that can be formulated for topical administration or delivered via other appropriate ocular routes. Such compositions can be used either as a monotherapy or in combination with other anti-inflammatory or anti-fibrotic agents to achieve comprehensive treatment outcomes. The present invention's method of treatment is applicable not only for acute ocular injuries but also for chronic conditions where both impaired healing and fibrosis are common, offering a significant advancement in the management of these conditions.

[0009] In an embodiment, through a dual-action mechanism, the present invention promotes epithelial regeneration and inhibiting fibrosis and addresses critical unmet needs in the treatment of ocular surface disorders. By enhancing both wound healing and tissue recovery while preventing scar formation, the present invention provides a comprehensive therapeutic strategy that can significantly improve patient outcomes in a variety of ocular pathologies.

[0010] The features and advantages of the subject matter here will become more apparent in light of the following detailed descriptionof selected embodiments, as illustrated in the accompanying FIGURES. As will be realized, the subject matter disclosed is capable of modifications in various respects, all without departing from the scope of the subject matter. Accordingly, the drawings and the description are to be regarded as illustrative in nature.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] In the figures, similar components and / or features may have the same reference label. Further, various components of the same type may be distinguished by following the reference label with a second label that distinguishes among the similar components. If only the first reference label is used in the specification, the description is applicable to any one of the similar components having the same first reference label irrespective of the second reference label.

[0012] FIG. 1 illustrates a flow chart for a method for IL-17c based therapeutics for promoting epithelial healing and inhibiting fibrosis in ocular disorders, in accordance with an embodiment of the present disclosure.

[0013] FIG. 2 illustrates a schematic representation of methodology of synthesis of lipid nanoparticle, in accordance with an embodiment of the present disclosure.

[0014] FIG.3A illustrates a schematic representation of complexation of RNA with ionic lipids, in accordance with an embodiment of the present disclosure.

[0015] FIG. 3B illustrates a schematic representation of RNA encapsulated LNP exhibit positive charge while at low pH and more neutral charge whenexposed to physiological pH, in accordance with an embodiment of the present disclosure.

[0016] FIG.4A illustrates patient cornea with PCED by fluorescein dye and diffuse light using a slit lamp, in accordance with an embodiment of the present disclosure.

[0017] FIG. 4B illustrates expression of IL17C at the mRNA level and at the protein level, in accordance with an embodiment of the present disclosure.

[0018] FIG. 5A illustrates 2D scratch assay to evaluate the migration of Human Comeal Epithelial Cells (HCLE), in accordance with an embodiment of the present disclosure.

[0019] FIG. 5B illustrates quantitative bar diagram of IL17C protein promoting epithelial migration and barrier function, in accordance with an embodiment of the present disclosure.

[0020] FIG. 5C illustrates establishment of ex-vivo model to evaluate effect of IL17C protein in wound healing, in accordance with an embodiment of the present disclosure.

[0021] FIG. 5D illustrates brief procedure to create ex vivo corneal model, in accordance with an embodiment of the present disclosure.

[0022] FIG. 5E illustrates effect of IL17C in promoting wound healing post debridement, in accordance with an embodiment of the present disclosure.

[0023] FIG. 6A illustrates reduction of alpha-SMA, in accordance with an embodiment of the present disclosure.

[0024] FIG. 6B illustrates prevention of collagen gel contraction, in accordance with an embodiment of the present disclosure.

[0025] FIG.6C illustrates validating antifibrotic potential of IL17C, in accordance with an embodiment of the present disclosure.

[0026] FIG. 7A-7B illustrate an in-vitro anti-angiogenesis assay on Matrigel, in accordance with an embodiment of the present disclosure.DETAILED DESCRIPTION OF THE DRAWINGS

[0027] The detailed description set forth below in connection with the appended drawings is intended as a description of exemplary embodiments in which the presently disclosed disclosure can be practiced. The term "exemplary" used throughout this description means "serving as an example, instance, or illustration," and should not necessarily be construed as preferred or advantageous over other embodiments. The detailed description includes specific details for providing a thorough understanding of the presently disclosed disclosure. However, it will be apparent to those skilled in the art that the presently disclosed disclosure may be practiced without these specific details. In some instances, well-known structures and devices are shown in block diagram form in order to avoid obscuring the concepts of the presently disclosed disclosure.

[0028] If the specification states a component or feature "may," "can," "could," or "might" be included or have a characteristic, that particular component or feature is not required to be included or have the characteristic.

[0029] As used in the description herein and throughout the claims that follow, the meaning of "a," "an," and "the" includes plural reference unless the context dictates otherwise. Also, as used in the description herein, the meaning of "in" includes "in" and "on" unless the context dictates otherwise.

[0030] The phrases "in an embodiment," "according to one embodiment," and the like generally mean the particular feature, structure, or characteristic following the phrase is included in at least one embodiment of the present disclosure and may be included in more than one embodiment of the present disclosure. Importantly, such phrases do not necessarily refer to the same embodiment.

[0031] It will be appreciated by those of ordinary skill in the art that the diagrams, schematics, illustrations, and the like represent conceptual views or processes illustrating systems and methods embodying this disclosure. The functions of the various elements shown in the figures may be provided through the use of dedicated hardware as well as hardware capable of executing associated software. Similarly, any switches shown in the figures are conceptual only. Their function may be carried out through the operation of program logic, through dedicated logic, throughthe interaction of program control and dedicated logic, or even manually, the particular technique being selectable by the entity implementing this disclosure. Those of ordinary skill in the art further understand that the exemplary hardware, software, processes, methods, and / or operating systems described herein are for illustrative purposes and, thus, are not intended to be limited to any particular name.

[0032] It is well known in the art that Herpes Simplex Keratitis (HSK) is one of the main causes of visual compromise around the world, usually as a result of recurrent Herpes Simplex Virus (HSV) infection of the cornea. While it is chiefly attributed to HSV type I, HSV type II can also be attributed to it, but commonly in newborns. These findings indicate global seroprevalence of 67% of individuals aged 0-49 years for HSV-1 and 11.3% for HSV-21. The HSK affects all age groups although its annual incidence of 10 to 30 per 100000 in developed countries is slightly higher in developing countries because of inadequate health care facilities and higher rates of co infection in children. The HSK is usually caused by reactivation of a latent HSV infection in the trigeminal ganglia. When activated, the virus goes to the cornea where it provokes an inflammatory response which results in corneal scarring and sometimes blindness2. Far advanced stromal keratitis can cause deeper penetration of the ulcer, currently, leading to permanent scar formation. As is clear, the HSK is a huge burden to patients and regardless of this, treatment efforts go more towards treating symptoms and working to prevent relapses. Although systemic antiviral agents that may be used include acyclovir or valacyclovir for epithelial and stromal keratitis, however, more work is needed to determine how each of the biomarkers under discussion reacts to the disease progress in human beings.

[0033] The IL-17C, of the IL- 17 cytokines family, involved in immune responses, more precisely inflammatory processes. It has an inflammatory effect as it combines with interleukin receptors 17RA and 17RE mostly on epithelial cells. IL17C stimulates synthesis of interleukins, especially IL6 and chemokines which play a role in pathogen resistance and inflammation, especially in ocular diseases, such as dry eye disease and keratitis3,4. Having increased and played a significant role in the activation of neutrophils and immune responses, IL-17C contributes toinflammation resulting in tissue damage in ocular infection such as bacterial and fungal keratitis. Further, in corneal fibrosis, it is observed that IL- 17 contributes and increase in fibroblast activation and of new components of the extracellular matrix (ECM), such as collagen type I and III necessary for the process of tissue repair4,5. However, when the activity continues for a long time, remodeling abnormally results in tissue scarring. It also controls matrix metalloproteinases (MMPs) thereby promoting the buildup of ECM. IL- 17 can be either anti-fibrotic as it leads to myofibroblast apoptosis, as well as pro-fibrotic at other stages of diseases since prolonged activation of fibroblasts occurs3,67. It also self-activates or modulates fibrosis through the cross- talk with the TGF-P pathway on myofibroblast differentiation. In addition to directly contributing to inflammation, IL- 17 is involved in angiogenesis during corneal fibrosis. It also generates pro-angiogenic cytokines such as VEGF and bFGF that provoke neo-vascularization in corneal stroma, fibroblasts are nourished by these new vessels resulting in augmented fibrosis. This angiogenesis generates a cycle in which vessel formation enhances fibroblast actions that exacerbate fibrous scarring and chronic inflammation7. Engagement with IL- 17 might present the possibility of therapeutic intervention aimed at reducing inflammation, fibroblast activation and preventing uncontrolled angiogenesis which affects corneal scarring and visual acuity. Though this is known in IL17 A and F but IL17C role is completely unknown in fibrosis, vascularization and corneal inflammation

[0034] In an embodiment, the present invention relates to IL-17C-based therapeutics designed for promoting epithelial healing and inhibiting fibrosis in ocular disorders. Such a therapeutic approach may be particularly beneficial in addressing conditions such as corneal ulcers, dry eye syndrome, and other inflammatory ocular surface disorders.Therapeutic Formulation

[0035] In an embodiment, the IL-17C-based therapeutic agent may include recombinant IL-17C protein or biologically active fragments of IL-17C, which may be derived from recombinant DNA technology. Further, the formulation may include additional excipients to enhance stability, bioavailability, and tolerability.The therapeutic agent may be configured as a topical solution, gel, or injectable formulation, ensuring ease of application and effective delivery to the ocular surface.Mechanism of Action

[0036] In an embodiment, the IL-17C may function through its receptor IL-17RE, which is expressed in epithelial cells and TH17 cells. In an example, the binding of IL-17C to IL-17RE may activate downstream signaling pathways that are involved in cellular proliferation and migration. This may promote re-epithelialization of the ocular surface by enhancing the wound-healing process.Promotion of Epithelial Healing

[0037] In an embodiment, the therapeutic agent may be configured to stimulate epithelial cell proliferation and migration. In an embodiment, the application of the IL-17C-based agent may lead to an increase in the expression of growth factors and cytokines that facilitate epithelial repair. The re-epithelialization process may include, but is not limited to, the activation of specific signaling pathways that promote cell growth and tissue regeneration.Inhibition of Fibrosis

[0038] In an embodiment, the IL-17C may also play a role in inhibiting fibrosis associated with ocular disorders. In an example, IL-17C may inhibit TGF-P activity, which is known to contribute to fibrotic processes. The therapeutic formulation may be designed to minimize collagen and fibronectin deposition in the affected ocular tissues. This may include mechanisms that downregulate the expression of genes involved in fibrosis and extracellular matrix remodeling.Administration Method

[0039] In an embodiment, the IL-17C-based therapeutic agent may be administered topically to the affected ocular surface, potentially in a specific dosage and frequency as determined by the treating clinician. In an embodiment, the method of administration may allow for targeted delivery to enhance therapeutic efficacy while minimizing systemic exposure.Monitoring and Assessment

[0040] In an embodiment, the present invention may include methods for monitoring the efficacy of the treatment. This may involve evaluating the healing progress of the ocular surface through clinical assessments, imaging techniques, and laboratory assays. In an example, monitoring may include tracking epithelial thickness, inflammation levels, and fibrosis markers over time to assess the therapeutic outcomes. The method has been discussed in detail in the following paragraphs.Long-term Effects

[0041] In an embodiment, the long-term effects of the IL-17C-based therapeutics may include sustained epithelial healing and reduced risk of scar formation. The therapy may be evaluated for its impact on visual acuity and quality of life in patients with ocular surface disorders.Potential Applications

[0042] In an embodiment, the IL-17C-based therapeutics may find applications beyond ocular disorders. In an example, the principles underlying this invention may be extended to treat other inflammatory diseases characterized by impaired wound healing and fibrosis, including, but not limited to, skin disorders, respiratory conditions, gastrointestinal diseases, Stevens-Johnson syndrome, vernal keratoconjunctivitis, and keratoconus, among others. Thus, the present invention presents a novel therapeutic approach that may leverage the unique properties of IL-17C to address significant challenges in the treatment of ocular surface disorders while also exploring potential applications in other areas of medicine.Materials and methodSample Collection

[0043] Epithelial scraping, a standard diagnostic and therapeutic procedure routinely employed for viral keratitis patients, was collected after taking informed consent. Scrapings taken from the leading edge of the ulcer were collected using a bard parker 15 no. blade under aseptic conditions, under the magnification of a slit lamp after the instillation of 0.5% proparacaine. When ulcer had healed, corneal impression cytology was collected using EYEPRIMTM. The samples werecollected in RNALater and immediately kept on ice. After collection, epithelial samples were stored at -20°C for further processing.

[0044] Similarly, the control corneal epithelial tissue were collected from individuals undergoing PRK surgery for the correction of refractive errors. The cornea was marked with an 8.5 mm trephine after which the corneal epithelium was debrided using a blunt spatula, under sterile conditions in the operating room under the magnification of an operating microscope after instillation of 0.5% proparacaine and preserved in RNA later for further processing. Alcohol-assisted epithelial debridement was not done. Tears from HSV1 patients as well as controls were collected with the Schirmer’s strips. They were transferred to a sterile 1.5 ml MCT and immediately kept on ice. After collection, tear samples were stored at -80°C for further processing.RNA isolation and qRT-PCR

[0045] RNA was isolated using the Qiagen Mini-RNA Isolation Kit (Cat. No.74004) as per the kit protocol. The isolated RNA was quantified using Nano Drop and stored at -80°C for further processing. Equal amount of RNA from HSV1 patients and control individuals was taken for cDNA preparation by using Revertaid First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, Cat. No. K1622) The reaction was incubated at 42°C for 60 minutes and then at 70°C for 5 minutes. The cDNA was stored at -20°C for further use.

[0046] Quantitative PCR (qRT-PCR) was performed to measure the gene expression. There actions were carried out in duplicate using SYBR Green dye on an Azureq RT-PCR instrument (Ceilo 6, Azure Biosystems, Dublin, CA, USA). EEF1A1 was used as the housekeeping gene for data normalization.The expression levels of the target genes were calculated relative to EEF1A1 using the AACt method.Results are presented as foldchanges. Plots for the analysis were generated using GraphPad Prism.Tear protein Isolation and quantification

[0047] Total protein was isolated from tear samples of HSV1 patients and controls using 300 pL of sterile IX PBS with IX protease inhibitor cocktail (PIC). Proteinquantification was performed using the BCA method with the Pierce BCA Protein Assay Kit (Cat.No.23227) following kit protocol. Equal amounts of protein from both patients and controls were used to quantify the levels of IL17c using BT Lab ELISA kits. The protein levels were assessed following the kit protocols. Data were analysed to determine the relative protein levels in patient samples compared to controls. Graphs illustrating this analysis were generated using GraphPad Prism.

[0048] Further, an O-link multiplex assay will be used to determine the cytokine milieu in tears obtained from patients with HSV-1 keratitis. This high proteomic technology permits multiple cytokines to be measured at once, giving information in relation to the inflammatory mediators of tear samples from patients with the disease.Cell culture experiments

[0049] In the cell culture studies we used human corneal limbal epithelial (HCLE) and isolated human corneal epithelial (HCE) cell lines. These different cell lines provide useful models for investigating corneal epithelial reactions because they closely reflect in vivo corneal tissue.

[0050] For the experiments, recombinant IL-17C protein was used to add exogenous treatment of the cultured cells. This strategy enables us to mimic the inflammatory context that might occur during the HSV-1 infection and directly evaluate the impact of IL-17C on cell response, viability, cytokine secretion, and intracellular signaling networks.

[0051] Furthermore, to achieve the development of the respective infected cell cultures, HSV-1 viral particles were added. This method allows, for example, studying the host cell response to viral infection when using IL- 17 C.Statistical Analysis

[0052] A Student's t-test was performed to analyse the data between control and patient samples. Standard error of the mean was used to plot the graph comparing controls versus patient samples. p<0.05 was considered statistically significant.ResultsqPCR of HSV1 keratitis patients epithelial tissue

[0053] We collected samples from HSV-1 keratitis patients during both the active stage, when the virus was actively present in the corneal epithelial tissue, and the resolved stage, after the ulcer had healed. For both stages, we performed qPCR to assess the gene expression of IL-17C. In the active stage, 22 patients with HSV-1 keratitis were analyzed, and we observed significantly elevated levels of IL-17C in the corneal epithelial tissue compared to control samples from photorefractive keratectomy (PRK) patients. This suggests that IL-17C plays a role in the inflammatory response during active HSV-1 infection. In the resolved stage, 32 patient samples were recruited for analysis. Interestingly, we observed a marked decrease in IL-17C expression in this stage compared to both the active stage and the control samples.

[0054] Downstream of the IL-17C signaling pathway, the regulation of the JAK / STAT pathway plays a crucial role. IL-6, a popular cytokine which has been linked to HSV-1 infection, fuctions through this same pathway. We have shown in the previous paper that there is a synergistic effect between IL-17C and IL-6 and the molecular crosstalk occurs through the STAT3 cascade. Based on this relationship, we examined the levels of IL-6 and STAT3 in both the active group of patients with HSV-1 keratitis and the resolved group of patients. Indeed, our data showed that the levels of IL-6 are much higher in the active stage as compared to the PRK control possibly implying the importance of this cytokine in modulating inflammation during active infection. Nonetheless, the level of IL-6 as a representative of inflammatory cytokines is significantly lower in the resolved stage of the disease. On the other hand, STAT3 expression did not decrease to PRK control levels in both active and resolved phases. The persistently high level of STAT3 that is observed here indicates its further active participation in the regulation of immune processes and regeneration of tissues after the termination of the virus replication and the acute formation of immune response.

[0055] Based on these results, we investigated the regulatory status of the two receptor subunits that are involved in IL-17C signaling in HSV-1 patients with keratitis. Our study showed that the transcripts of both receptors were upregulatedduring the active phase of the disease. This seen increase infers that the IL-17C signaling pathway is intense during the early days of HSV-1 infection, eliciting the severe inflammato- pared to the normal conjunctiva. Such upregulation of both IL-17 receptors probably helps to enhance the synthesis of pro-inflammatory cytokines, accumulation of immune cells, and stromal remodeling, which represents the main pathogenetic signs of HSV-1 keratitis. The level of IL-17RA and IL-17RE was significantly lower in the resolved stage than in the active stage of the wound.Tear protein factors evaluated by ELISA

[0056] Tear samples were obtained from patients with HSV-1 keratitis both in the active phase of the disease and from non-infectious control patients. In the current study the quantification of IL-17C protein in these tear samples showed a substantial increase in active stage HSV-1 keratitis patients as compared to controls. Much higher levels of IL- 17C were measured in the active stage of infection, which points towards the involvement of this cytokine in directing the inflammation process in the infected corneal epithelial tissue. However, when comparing the levels of IL-17C protein in the resolved stage with active stage and control tears, substantial decrease of the score was detected. They found that the decrease in IL-17C during the healing phase indicates anti-inflammatory rhythms and promotes tissue repair and remodeling.Tear profiling by O-linkCell culture experiment

[0057] In this study, human corneal limbal epithelial (HCLE) and human corneal epithelial (HCE) cell lines were infected with HSV-1 viral particles at a 102dilution. Finally, following 6 hours, there was a sustained increase in the expression of IL-17C in both cell lines. This rapid increase indicates that it plays an important part in the initial immune response to HSV-1 infection, a process most probably associated with the recruitment of immune cells to the site of infection and resultant inflammation. It may therefore be important for IL-17C to be induced early enoughto control the immune environment in HSV-1 keratitis as a possible therapeutic target to prevent inflammation and tissue degradation.

[0058] FIG. 1 illustrates a flow chart 100 for a method for IL- 17c based therapeutics for promoting epithelial healing and inhibiting fibrosis in ocular disorders, in accordance with an embodiment of the present disclosure. The method starts at step 102.

[0059] At first, the ocular surface disorder may be identified, at step 104. Such identification may include a thorough clinical examination to diagnose conditions such as corneal ulcers, dry eye syndrome, or inflammatory ocular disorders. Further, such identification may be performed by utilizing various diagnostic imaging techniques and evaluations, including slit-lamp examinations, to determine the extent of epithelial damage and inflammation. By accurately diagnosing the specific ocular surface disorder, clinicians may tailor subsequent therapeutic interventions effectively, ensuring that the treatment strategy aligns with the unique needs of the patient and the characteristics of the disorder being treated.

[0060] Next, at step 106, the IL-17C-based therapeutic agent may be prepared and formulated, such as, by developing a suitable form of IL- 17C, such as a recombinant protein or biologically active fragment, which is designed for effective delivery to the ocular surface. The formulation may be created as a topical solution, gel, or injectable, prioritizing stability and bioavailability to ensure efficient absorption into the eye. Additionally, this step may include optimizing the composition with excipients that enhance the therapeutic agent's stability, prolong its shelf life, and minimize irritation to the sensitive ocular tissues, ultimately ensuring a safe and effective therapeutic option for patients.

[0061] Next, at step 108, epithelial healing may be promoted by designing the therapeutic application regimen to encourage epithelial cell regeneration and incorporating instructions for the administration frequency and dosage of the IL-17C-based therapeutic agent. Next, at step 110, a therapeutic agent may be administered by applying the IL-17C-based therapeutic agent directly to the injured ocular surface as prescribed and using sterile techniques to prevent contaminationduring the administration process. Next, at step 112, epithelial cell proliferation and migration may be stimulated by allowing the IL-17C agent to act on the epithelial cells, initiating an autocrine signaling cascade that promotes cell proliferation and migration and monitors cellular responses, such as increased epithelial thickness and healing rates, through clinical assessments and imaging techniques.

[0062] Next, at step 114, re-epithelialization and wound healing may be performed by assessing the progress of re-epithelialization through follow-up examinations at regular intervals and evaluating the restoration of the epithelial barrier and the resolution of inflammation, as indicated by symptom relief and improved ocular surface stability. Next, at step 116, inhibition of fibrosis may be performed by concurrently monitoring for signs of fibrosis in the ocular tissue through the use of IL-17C’s properties to inhibit TGF-P signaling pathways, which are known to contribute to fibrotic processes. Next, at step 118, TGF-P activity may be assessed by evaluating the effectiveness of the IL-17C-based agent in inhibiting TGF-P activity through laboratory assays (e.g., enzyme-linked immunosorbent assay (ELISA) for TGF-P levels) and histological analysis of the ocular tissue. Next, at step 120, collagen and fibronectin deposition may be reduced and changes in the levels of collagen and fibronectin deposition may be monitored in the treated area through tissue analysis and imaging studies while the balance of extracellular matrix components may be assessed to ensure minimal fibrosis and optimal healing. Thereafter, at step 122, fibrosis and scar formation may be prevented such as, by conducting long-term follow-up to ensure sustained healing and prevention of scar formation and assessing visual acuity and quality of life improvements in patients receiving the IL-17C-based therapeutic regimen. The method ends at step 124.

[0063] FIG. 2 illustrates a schematic representation 200 of methodology of synthesis of lipid nanoparticle, in accordance with an embodiment of the present disclosure. The workflow commences by providing cholesterol in CHCL / MeOH as shown at 202 together with one or more lipids such as HSPC, DOPC, DOTAP and / or DOPE in CHCL / MeOH as shown at 206, which are mixed in anequimolar ratio as indicated at 204 to form a homogeneous pre-liposome solution.

[0064] In an embodiment, the pre-liposome solution is subjected to drying by rotary evaporation and / or vacuum desiccation as depicted at 220 so as to remove organic solvent and promote thin-film formation on a vessel surface as indicated at 208. The thin film 208 is then hydrated with phosphate-buffered saline as shown at 210 to form a multilamellar dispersion. In some implementations, the hydrated film is dislodged by brief water-bath sonication, for example about one minute, as shown at 222 to facilitate lamellae separation and prepare the dispersion for sizing.

[0065] In an embodiment, the hydrated dispersion is sized by extrusion as indicated at 212 through polycarbonate membranes using a mini-extruder as shown at 224 to obtain different sized liposomes as represented at 214.Selection of membrane pore sizes and number of extrusion passes may be used to tailor the particle size distribution for subsequent loading of IL-17C mRNA or IL-17C protein and for evaluation in ocular delivery screens.

[0066] In an embodiment, the obtained suspensions are characterized as indicated at 216 using one or more analytical techniques such as dynamic light scattering, nanoparticle tracking analysis and zeta-potential measurement to confirm hydrodynamic size, distribution and surface charge appropriate for the intended formulation profile. Batches meeting the desired criteria are stored at approximately 4 °C as shown at 218 for use in subsequent in-vitro or ex-vivo studies.

[0067] In an embodiment, cargo association is carried out within the same workflow 200 by hydrating the thin film 208 with an aqueous phase comprising IL-17C mRNA or by permitting passive entrapment of IL-17C protein during the hydration 210 and sonication 222 stages, thereby producing vesicles suitable for biological evaluation. Process parameters such as mix time at 204, evaporation endpoint at 220, hydration volume at 210, sonication interval at 222, and number of extrusion cycles at 212 with the mini-extruder 224 may be recorded and correlated with characterization outputs at 216 to supportreproducible preparation.

[0068] In an embodiment, the illustrated sequence of steps is implemented to generate IL-17C-bearing lipid systems that enable investigations aligned to program objectives. In particular, preliminary research motivating this workflow suggests that IL-17C may participate in aspects of corneal homeostasis and could be relevant to persistent corneal epithelial defect and fibrosis, while the precise impact remains under study. Accordingly, Objective 1 contemplates validating IL-17C expression in patient samples by qPCR, ELISA or immunohistochemistry, and Objective 2 contemplates developing an IL-17C mRNA delivery system using the formulations prepared by the methodology of FIG. 2 for evaluation in corneal epithelial cell models and appropriate exploratory model systems.

[0069] FIG. 3A illustrates a schematic representation 300A of complexation of RNA with ionic lipids, in accordance with an embodiment of the present disclosure.FIG. 3B illustrates a schematic representation 300B of RNA encapsulated LNP exhibit positive charge while at low pH and more neutral charge when exposed to physiological pH, in accordance with an embodiment of the present disclosure. For the sake of brevity, FIGs. 3A and 3B have been explained together.

[0070] In an embodiment, FIG. 3A illustrates a schematic representation 300A of a lipid bilayer vesicle configured to associate and protect a polyanionic RNA cargo. The bilayer includes a population of complexing or ionizable lipids distributed in the inner and outer leaflets to promote electrostatic association with the RNA during assembly, together with helper lipids such as DOPE that facilitate membrane curvature and endosomal escape, and cholesterol that modulates membrane packing and stability. The vesicle is drawn with an interior aqueous compartment and an exterior aqueous phase to depict that the RNA can be located within the internal lumen and / or intercalated at the inner leaflet, while the distribution of the lipids across the two leaflets is representative and not limiting. In certain implementations, a surface-presented PEG-lipid may be included (not drawn to scale in FIG. 3A) to provide colloidal stability and to modulate interfacial interactions during ocularformulation handling.

[0071] In an embodiment, FIG. 3B illustrates a schematic representation 300B of charge behavior for RNA-encapsulated lipid nanoparticles under differing pH conditions encountered during manufacturing and use. At acidic or assembly pH, cationic or ionizable lipids are predominantly protonated, yielding a net positive surface character that strengthens complexation with the RNA and assists encapsulation within the particle. The helper lipid and cholesterol contribute to membrane structure, while PEG-lipid forms a hydrated corona that improves dispersion. Upon exposure to near-physiological pH, the ionizable lipids become less protonated, and the particle exhibits a more neutral apparent surface character while maintaining association with the RNA within the vesicle interior. The transition from the positive character at low pH to the more neutral character at physiological pH is depicted conceptually to indicate that charge state is a function of the ionization of lipids and the surrounding medium and can be tuned by lipid selection and formulation conditions.

[0072] In an embodiment, the arrangements shown in FIGS. 3A and 3B are exemplary and may be implemented with different mole ratios of complexing lipids, helper lipids, cholesterol, and PEG-lipids to balance encapsulation efficiency, particle integrity, and dispersion stability. The size of the vesicle and the length of PEG chains are not to scale and are provided to indicate relative placement only. Without being bound by theory, the inclusion of helper lipids such as DOPE can favor membrane fusion or destabilization events that, together with the pH-responsive ionizable lipids, may facilitate cytoplasmic access of the RNA after cellular uptake.

[0073] FIG. 4A illustrates patient cornea 400A with PCED by fluorescein dye and diffuse light using a slit lamp, in accordance with an embodiment of the present disclosure. FIG. 4B illustrates expression 400B of H 17C at the mRNA level and at the protein level, in accordance with an embodiment of the present disclosure. For the sake of brevity, FIGs. 4A and 4B have been explained together.

[0074] In an embodiment, FIG. 4A illustrates patient cornea 400A documented on a slit lamp for a case of persistent corneal epithelial defect (PCED). The upper-leftpanel shows cobalt-blue illumination with fluorescein instillation demonstrating pooling and edge staining consistent with a non-healing epithelial defect; the upperright panel shows the same eye under diffuse white light highlighting stromal haze and surface irregularity. The lower-left panel depicts a subsequent fluorescein view again showing linear / geographic staining along the defect margins, while the lower-right panel provides a companion diffuse-light view indicating residual opacity and surface disturbance. The photographs were acquired with standard clinical settings on a slit lamp using fluorescein dye and cobalt -blue excitation for the fluorescein panels, and diffuse illumination for the companion panels. No patient identifiers are shown; image magnification and color balance are illustrative and not limiting. The panels collectively depict typical clinical features associated with PCED, namely persistent epithelial staining, irregular epithelium, and variable stromal haze and opacity.

[0075] In an embodiment, FIG. 4B illustrates expression analysis 400B of IL-17C performed on clinical samples obtained from a PCED cohort and from control subjects. The left graph presents messenger RNA measurements for IL-17C using a quantitative polymerase chain reaction assay expressed as relative fold change; individual data points from control specimens and from samples obtained during an active disease stage are plotted with a central tendency indicator and, where shown, dispersion bars. The right graph presents protein measurements for IL-17C obtained by an enzyme-linked immunosorbent assay and reported in ng / mL, again displaying individual values for control and active-stage groups with corresponding central tendency and dispersion. The dotted reference line (if present) denotes an analytical baseline or normalization level used in the assay pipeline. Without limiting the disclosure to any particular numerical outcome, the plotted distributions in FIG.4B are representative of a trend in which higher IL-17C readouts may be observed in samples collected during an active disease stage relative to control samples. The data are preliminary and provided to illustrate the analytical framework that informs the invention; sample sizes, assay parameters, and statistical treatments can vary according to laboratory protocol.

[0076] FIG. 5A illustrates 2D scratch assay 500A to evaluate the migration ofHuman Comeal Epithelial Cells (HCLE), in accordance with an embodiment of the present disclosure. FIG. 5B illustrates quantitative bar diagram 500B of IL17C protein promoting epithelial migration and barrier function, in accordance with an embodiment of the present disclosure. FIG.5C illustrates establishment of ex-vivo model 500C to evaluate effect of IL17C protein in wound healing, in accordance with an embodiment of the present disclosure. FIG. 5D illustrates brief procedure 500D to create ex vivo comeal model, in accordance with an embodiment of the present disclosure. FIG. 5E illustrates effect 500E of IL17C in promoting wound healing post debridement, in accordance with an embodiment of the present disclosure. For the sake of brevity, FIGs. 5A-5E have been explained together.

[0077] In an embodiment, FIG.5A depicts a 2D scratch assay 500A conducted with human corneal epithelial cells to visualize epithelial migration over time. Representative micrographs are shown at the starting time point (0 hours) and after an incubation interval, with the denuded “scratch” area outlined to indicate the wound margin. The vehicle-treated control frames demonstrate partial closure of the defect, whereas the frames corresponding to IL-17C protein exposure (for example, 50 ng / mL) show visibly greater advance of the epithelial front into the scratched region, consistent with enhanced migratory activity under IL-17C.

[0078] In an embodiment, FIG. 5B presents a quantitative bar diagram 500B summarizing the effect of IL-17C on epithelial migration and barrier function. The plotted measure reflects percentage wound closure calculated from image analysis of the scratch assay at a fixed time point. The bar corresponding to the IL-17C condition exceeds that of the control, and a significance indicator is shown over the paired bars, illustrating that IL-17C treatment is associated with increased closure versus untreated cells under the assay conditions.

[0079] In an embodiment, FIG. 5C illustrates the establishment of an ex-vivo model 500C for evaluating the effect of IL-17C protein on corneal wound healing. The depiction shows corneal tissue mounted and prepared for standardized debridement, followed by exposure to test and control preparations. This model provides a controlled platform to assess epithelialrestoration and surface integrity after injury while maintaining native tissue architecture.

[0080] In an embodiment, FIG. 5D summarizes a brief procedure 500D to create the ex-vivo corneal model. The sequence shows corneal isolation, epithelium debridement, placement into holders or chambers, and subsequent allocation for downstream analyses such as gene-expression studies, hematoxylin-and-eosin staining, and immunohistochemistry. Arrows in the sequence indicate the order of operations from tissue preparation through to analytical readouts.

[0081] In an embodiment, FIG. 5E shows the effect 500E of IL-17C in promoting wound healing after debridement in the ex-vivo model. Comparative histology panels (for example, hematoxylin-and-eosin and periodic acid-Schiff stains) display the epithelial surface and stromal interface in control tissue versus tissue exposed to IL-17C. The IL-17C panels exhibit a more continuous superficial epithelial layer and a smoother epithelial-stromal boundary relative to control, consistent with improved re-epithelialization under the test condition.

[0082] FIG. 6A illustrates reduction of alpha-SMA 600A, in accordance with an embodiment of the present disclosure. FIG. 6B illustrates prevention of collagen gel contraction 600B, in accordance with an embodiment of the present disclosure.FIG. 6C illustrates validating antifibrotic potential of IL17C 600C, in accordance with an embodiment of the present disclosure. For the sake of brevity, FIGs. 6A-6C have been explained together.

[0083] In an embodiment, FIG. 6A illustrates reduction of alpha-SMA 600A using a corneal fibroblast assay imaged under phase-contrast microscopy. Representative fields are shown for basal conditions, for exposure to IL-17C alone, for stimulation with TGF-pi, and for co-exposure to TGF-pi together with IL-17C. The TGF-pi panel shows organized stress-fib er-like features and elongated morphology consistent with myofibroblast transition, whereas the co-exposure panel displays a visibly attenuated stress-fiber appearance and a return toward a less contractile morphology relative to TGF-pi alone. The basal and IL-17C-alone panels exhibitcomparatively fine cytoskeletal texture. Collectively, these images depict a qualitative decrease in alpha-SMA-associated features in the presence of IL-17C under the illustrated conditions.

[0084] In an embodiment, FIG. 6B illustrates prevention of collagen gel contraction 600B using a three-dimensional gel assay photographed from above at two time points. Gels maintained under control conditions retain a larger diameter over time, whereas gels stimulated with TGF-pi exhibit pronounced circumferential contraction, evidenced by a reduced gel footprint. Gels exposed to IL-17C alone show minimal contraction similar to control, and gels co-exposed to IL-17C with TGF-pi maintain a visibly larger area than TGF-pi alone at both time points. The side-by-side series demonstrates that inclusion of IL-17C is associated with preservation of gel size, indicating reduced contractile remodeling in this model.

[0085] In an embodiment, FIG. 6C illustrates validating antifibrotic potential of IL-17C 600C using a quantitative readout of alpha-SMA. The plotted bars summarize relative alpha-SMA signal for control, IL-17C, TGF-pi, and TGF-pi plus IL-17C conditions. The TGF-pi bar is elevated compared to control, consistent with profibrotic induction, whereas the bar for TGF-pi plus IL-17C is reduced relative to TGF-pi alone. The IL-17C-alone bar is near the control level. Without limiting the disclosure to any particular numerical value, the pattern supports the qualitative observations in the microscopy and gel assays and is consistent with an antifibrotic effect associated with IL-17C under the illustrated conditions.

[0086] FIG. 7A-7B illustrate an in-vitro anti-angiogenesis assay 700A, 700B on Matrigel, in accordance with an embodiment of the present disclosure.

[0087] In an embodiment, FIG. 7A illustrates an in-vitro anti-angiogenesis assay 700A performed on Matrigel using endothelial network formation as a readout. Representative image panels are arranged by condition, media alone (control), IL-17C at 50 ng, and IL-17C at 100 ng, and by imaging mode, with phase-contrast micrographs captured at approximately 4xmagnification and companion fluorescence micrographs captured at approximately 1 Oxmagnification. Under media alone, contiguous capillary-like meshes and polygonal lattices are evident.In the presence of IL-17C at 50 ng, the meshes appear more fragmented with fewer interconnections and thinner cords across multiple fields. At 100 ng, the fluorescent images further illustrate sparse, interrupted segments and reduced nodal interlinks relative to control. The panels together depict a qualitative reduction in network complexity with IL-17C under the illustrated assay conditions.

[0088] In an embodiment, FIG. 7B presents quantification 700B of the networkformation assay corresponding to the imaging in FIG. 7A. The upper scatter plot summarizes the number of junctions per field, while the lower scatter plot summarizes total segment length per field; individual data points are shown for media alone, IL-17C 50 ng, and IL-17C 100 ng, with central tendency and dispersion indicators. Relative to media alone, both the number of junctions and the total segment length are reduced in cultures exposed to IL-17C, with the effect evident across biological replicates in the data displayed. Without limiting the disclosure to any particular numerical outcome, the plotted trends are consistent with inhibition of tube-like network formation in the presence of IL-17C in this Matrigel assay.

[0089] In an embodiment, the present invention relates to identifying marker of ocular surface diseases. The present invention particularly relates to an in-vitro method of identifying biomarker associated with one or more ocular surface diseases, such as for determining one or more ocular surface diseases, in a subject. Moreover, the present invention also particularly relates to a point-of-care device for identifying biomarker associated with one or more ocular surface diseases. The principal object of the present invention is to provide a solution to the existing problem of inaccurate determination (for the purposes of diagnosis, monitoring and management) of one or more ocular surface diseases by providing a novel in-vitro method for a spectrum of one or more ocular surface diseases, spanning a diverse array of conditions impacting the ocular surface, including, but not restricted to, Ocular fibrosis, keratoconus (KC), vernal keratoconjunctivitis (VKC), KC with VKC, KC with allergies, dry eye syndrome, conjunctivitis, keratitis, corneal dystrophies, and ocular surface secondary to mechanical, chemical, infection, biological and environmental injury, by detecting and quantifying expression levelsof IL17C and IL6 gene expression and proteins, within ocular samples, as diagnostic markers and therapeutic targets for one or more ocular surface diseases.

[0090] This invention claims for recombinant IL17C (R-IL17C), either alone or in combination with other protein, chemical and biologies, or in conjugation other molecule such as chemical conjugates, peptide, nanobodies, scFvs, encapsulate in nanoparticle, nanovesicles, exosomes for the below mentioned conditions.

[0091] Pro-regeneration effect of recombinant IL17C (R-IL17C) on ocular surface and corneal epithelium secondary to mechanical, chemical, infection, biological and environmental injury.

[0092] Anti-fibrotic effect of R-IL17C in ocular surface and ocular tissue secondary to above mention condition.

[0093] Anti-fibrotic effect of R-IL17C due to TGF-beta expression in ocular surface and other tissues.

[0094] Anti-angiogenic effect of R-IL17C in ocular surface and ocular tissues.

[0095] Pro-neurogenic effect of R-IL17C in neurotrophic keratitis, ocular surface and other ocular tissues.

[0096] R-IL17C as a modulator of Extracellular matrix (ECM) in the above-mentioned conditions.

[0097] In an embodiment, the present invention offers several advantages in the treatment of ocular surface disorders through the use of IL-17C-based therapeutics. Firstly, it may promote effective re-epithelialization and enhance wound healing, addressing a critical need in conditions like corneal ulcers and dry eye syndrome where rapid recovery of the epithelial layer is essential. Additionally, by inhibiting TGF-P activity, the therapeutic agent may reduce fibrosis and minimize collagen and fibronectin deposition, potentially preventing long-term complications associated with scarring and tissue remodeling. The targeted delivery of IL-17C may allow for localized treatment, thereby reducing the risk of systemic side effects commonly associated with broader therapeutic approaches. Furthermore, the antiinflammatory properties of IL-17C may provide a dual benefit, not only alleviating symptoms but also modulating the immune response to enhance overall ocular health. Overall, the innovative approach of utilizing IL-17C for therapeuticintervention may significantly improve patient outcomes and quality of life for individuals suffering from various ocular surface disorders.

[0098] While the embodiments of the invention have been described in detail for illustrative purposes, it is important to note that these examples are not intended to limit the scope of the invention. Various modifications, alterations, and adaptations may be made by those skilled in the art without departing from the spirit and scope of the invention. The invention encompasses all such variations and is not restricted to the specific embodiments presented herein.

Claims

CLAIMSWe Claim:

1. A pharmaceutical composition for ocular administration, the pharmaceutical composition comprising a therapeutic agent selected from:(a) interleukin- 17C (IL-17C) protein;(b) messenger RNA (mRNA) encoding IL-17C; and(c) IL-17C protein encapsulated in a delivery vehicle;wherein the therapeutic agent is formulated with a delivery system selected from lipid nanoparticles (LNPs), exosomes, hydrogels, and ocular inserts.

2. The pharmaceutical composition as claimed in claim 1, wherein when evaluated in ocular models, the pharmaceutical composition reduces fibrosis and neovascularization, promotes epithelial regeneration, and enhances corneal nerve repair.

3. The pharmaceutical composition as claimed in claim 1, wherein the therapeutic agent is at least one of:recombinant IL-17C protein formulated for topical ocular use; and IL-17C mRNA encapsulated in LNPs or exosomes.

4. The pharmaceutical composition as claimed in claim 1, wherein the IL-17C protein is encapsulated in LNPs or exosomes for sustained release.

5. The pharmaceutical composition as claimed in claim 1 , wherein the exosomes are derived from mesenchymal stem cells, corneal epithelial cells, or dendritic cells, and are loaded with IL-17C mRNA or IL-17C protein.

6. The pharmaceutical composition as claimed in claim 1, wherein the LNP formulation comprises an ionizable lipid, cholesterol, a phospholipid, and a PEG-lipid.

7. The pharmaceutical composition as claimed in claim 1, wherein the LNPs have a particle size of 60-120 nm and a near-neutral zeta potential.

8. The pharmaceutical composition as claimed in claim 1, wherein the IL-17C mRNA includes chemically modified nucleotides selected frompseudouridine and 5-methyl cytidine, and further comprises a 5' cap, optimized untranslated regions (UTRs), and a poly(A) tail.

9. The pharmaceutical composition as claimed in claim 1, whereinwhen evaluated in corneal fibroblast assays, the pharmaceutical composition reduces a-SMA expression and collagen gel contraction;when evaluated in a rabbit alkali burn model, the pharmaceutical composition decreases corneal opacity and neovascularization by at least 40- 50% compared to untreated controls;when evaluated in corneal debridement models, the pharmaceutical composition accelerates wound closure by at least 30% compared to untreated controls; andwhen evaluated following corneal nerve injury, the pharmaceutical composition restores corneal nerve density and / or sensitivity by at least 25% compared to baseline.

10. A sustained-release ocular device comprising a pharmaceutical composition for ocular administration, the pharmaceutical composition comprising a therapeutic agent selected from:(a) interleukin- 17C (IL-17C) protein;(b) messenger RNA (mRNA) encoding IL-17C; and(c) IL-17C protein encapsulated in a delivery vehicle;wherein the therapeutic agent is formulated with a delivery system selected from lipid nanoparticles (LNPs), exosomes, hydrogels, and ocular inserts;wherein the sustained-release ocular device is selected from hydrogel implants, corneal bandage contact lenses, and biodegradable ocular inserts.

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