Compositions and methods for treating pterygium

Abstract

The present invention provides compositions and methods for inducing pterygium regression from the visual axis / corneal center, stabilizing pterygium, treating hyperemia and symptoms in pterygium patients, and treating pterygium recurrence after pterygium excision. [Solution] The method involves administering to the patient a multi-kinase inhibitor, an antimetabolite, or a combination thereof.

Description

[Technical Field]

[0001] This disclosure relates to ophthalmic compositions and methods for treating primary and recurrent pterygium, more specifically, compositions and methods for inducing pterygium regression from the visual axis / corneal center, stabilizing pterygium, and suppressing ocular conjunctival hyperemia and pterygium recurrence before, during, or after pterygium surgery. [Background technology]

[0002] Pterygium is an ocular surface disease characterized by abnormal proliferation of epithelial cells and fibroblasts that extends from the nasal or temporal conjunctiva, across the limbus, and into the cornea. Patients with pterygium often experience eye discomfort and redness, and if the lesion invades the visual axis, there is a risk of causing visual impairment. [Overview of the project] [Means for solving the problem]

[0003] In certain embodiments, the disclosure provides a method for treating primary and recurrent pterygium by administering (1) a multi-kinase inhibitor, (2) an antimetabolite, or (3) a combination of a multi-kinase inhibitor and an antimetabolite to the eye of a subject requiring treatment. In certain embodiments, the method disclosed herein stabilizes the pterygium and prevents further proliferation of the affected tissue. In other embodiments, the method disclosed herein induces pterygium regression from the optic axis / corneal center. In certain embodiments, the multi-kinase inhibitors disclosed herein target the kinase receptors of VEGFR(1,2,3) and PDGFR(α,β). In certain embodiments, the multi-kinase inhibitor is incorporated into a topical ophthalmic formulation for topical administration to the affected eye. In certain embodiments, the topical ophthalmic formulation is a solution, suspension, or emulsion. In other embodiments, the multi-kinase inhibitor is incorporated into an implant or semi-solid sustained-release formulation for insertion into the affected eye. In certain embodiments, the antimetabolite is mitomycin C, 5-fluorouracil, and thiotepa. In a particular embodiment, the antimetabolite is incorporated into a topical ophthalmic formulation administered locally to the affected eye. In a particular embodiment, the topical ophthalmic formulation is a solution, suspension, or emulsion. In another embodiment, the antimetabolite is incorporated into an implant or semi-solid sustained-release formulation inserted into the affected eye. In a particular embodiment, the method disclosed herein is carried out by a combination of a multi-kinase inhibitor and an antimetabolite. In a particular embodiment, the combination of a multi-kinase inhibitor and an antimetabolite is incorporated into a topical ophthalmic formulation administered locally to the affected eye. In a particular embodiment, the topical ophthalmic formulation is a solution, suspension, or emulsion. In another embodiment, the combination of a multi-kinase inhibitor and an antimetabolite is incorporated into an implant or semi-solid sustained-release formulation inserted into the affected eye. In a particular embodiment, the method disclosed herein alleviates hyperemia, abnormal neovascularization, and other symptoms in pterygium patients. In another embodiment, the method disclosed herein prevents pterygium recurrence after pterygium surgery. In certain embodiments, the methods disclosed herein are performed before, concurrently with, or after surgical removal of a pterygium to suppress or prevent pterygium recurrence.In certain embodiments, the multikinase inhibitor disclosed herein targets a minimum number of kinase receptors, including VEGFR(1, 2, 3), PDGFR(α, β), and FGFR(1, 2, 3, 4), as well as optimal kinase receptors, including FLT3, Lck, Lyn, and Src. In certain embodiments, the multikinase inhibitor disclosed herein is nintedanib. In certain embodiments, the method disclosed herein uses a topical ophthalmic formulation. In certain embodiments, the formulation is an aqueous solution, suspension, or emulsion. In certain embodiments, the concentration of nintedanib in the formulation is 0.001% to 10%. In certain embodiments, the formulation is an implant inserted into the affected eye or a semi-solid sustained-release formulation. In certain embodiments, the amount of nintedanib in the implant is 1 μg to 100 mg.

[0004] In one embodiment, the Disclosure provides a method for inducing pterygium regression from the visual axis / corneal center, comprising administering a therapeutically effective amount of (1) a multi-kinase inhibitor; (2) an antimetabolite that inhibits epithelial and fibroblast proliferation; or (3) a combination thereof to the affected eye of a subject requiring such treatment. In a predetermined embodiment of the entire spectrum, the administration of the multi-kinase inhibitor, the antimetabolite, or a combination thereof results in a reduction in the size of the pterygium in the affected eye. In some cases, the administration of the multi-kinase inhibitor, the antimetabolite, or a combination thereof results in a negative pterygium growth rate in the affected eye.

[0005] In another embodiment, the Disclosure provides a method for stabilizing a pterygium, comprising administering a therapeutically effective amount of (1) a multi-kinase inhibitor; (2) an antimetabolite that inhibits epithelial and fibroblast proliferation; or (3) a combination thereof to the affected eye of a subject requiring such treatment. In some cases, as a result of the administration of the multi-kinase inhibitor, the antimetabolite, or a combination thereof, the size of the pterygium in the affected eye is stabilized. In some cases, as a result of the administration of the multi-kinase inhibitor, the antimetabolite, or a combination thereof, the rate of pterygium growth in the affected eye becomes nearly zero.

[0006] In all predetermined embodiments, the multikinase inhibitor reduces the activity of one or more intracellular and / or cell surface protein kinases selected from EGFR, ErbB2, ErbB3, FGFR1, FGFR2, FGFR3, FGFR4, TrkA, NGFR, VEGFR(1, 2, 3), PDGFR(α, β), TGF-βR(I, II, III), FLT3, Lck, Lyn, Src, c-Kit, c-Fms, Raf-1, B-Raf, RET, and CSF-1R in the pterygium. The multikinase inhibitor may have an IC50 <200 nM for VEGFR(1, 2, 3), an IC50 <200 nM for PDGFR(α, β), and / or an IC50 <1000 nM for FGFR(1, 2, 3).

[0007] In a predetermined embodiment of the entire set, the multikinase inhibitor is afatinib, amvatinib, axitinib, cabozantinib, canertinib, cedilanib, ceritinib, clenolanib, crizotinib, dabrafenib, dacomitinib, dasatinib, erlotinib, foretinib, gefitinib, golbatinib, ibrutinib, icotinib, idelalisib The antimetabolite is selected from the group consisting of imatinib, lapatinib, lenvatinib, neratinib, nilotinib, nintedanib, palbociclib, pazopanib, ponatinib, quizartinib, regorafenib, ruxolitinib, sorafenib, sunitinib, tanzutinib, tivantinib, tivozanib, trametinib, vandetanib, batalanib, and vemurafenib. In some cases, the antimetabolite is selected from the group consisting of mitomycin C, 5-fluorouracil, floxuridine, cytarabine, 6-azauracil, azathioprine, methotrexate, mycophenolate mofetil, and thiotepa.

[0008] In all of the specified embodiments, the multi-kinase inhibitor, the antimetabolite, or a combination thereof is administered to the affected eye in the form of a topical ophthalmic preparation, ointment, gel, sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant. If necessary, the multi-kinase inhibitor, the antimetabolite, or a combination thereof is administered to the affected eye in the form of a topical ophthalmic preparation, or administered locally to the affected eye.

[0009] Depending on the circumstances, the topical ophthalmic preparation may be a solution, suspension, or emulsion. Depending on the circumstances, the topical ophthalmic preparation may further contain one or more pharmaceutically acceptable additives selected from stabilizers, surfactants, polymer-based carriers, gelling agents, organic auxiliary solvents, pH adjusting components, and osmotic pressure adjusting components, and may or may not contain preservatives. Depending on the circumstances, the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant is inserted into the affected eye. In a given embodiment, the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant may further contain pharmaceutically acceptable additives. Depending on the circumstances, the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant may contain a multi-kinase inhibitor, an antimetabolite, or a combination thereof; and a biodegradable polymer selected from polylactic acid (PLA), polyglycolic acid (PLGA), and copolymers of polylactic acid and polyglycolic acid.

[0010] In all of the specified embodiments, the administration is performed on a patient with pterygium.

[0011] In another embodiment, the present disclosure provides a method for reducing hyperemia and symptoms in pterygium, pinguecula, and pseudopterygium in patients requiring treatment, comprising administering a therapeutically effective dose of a multikinase inhibitor to the affected eye of the subject.

[0012] In another embodiment, the Disclosure provides a method for reducing or preventing pterygium recurrence in subjects requiring treatment, comprising administering a therapeutically effective dose of a multikinase inhibitor to the affected eye of the subject. In all predetermined embodiments, the administration is performed before surgical removal of the pterygium. Optionally, the administration is performed during the surgical removal procedure. Optionally, the administration is performed after the surgical removal of the pterygium.

[0013] In a predetermined embodiment of the entire set, the multi-kinase inhibitor has an IC50 of <50 nM for VEGFR(1, 2, 3), an IC50 of <100 nM for PDGFR(α, β), an IC50 of <150 nM for FGFR(1, 2, 3), an IC50 of <1000 nM for FGFR4, an IC50 of <50 nM for FLT3, an IC50 of <50 nM for Lck, an IC50 of <200 nM for Lyn, and an IC50 of <200 nM for Src. In some cases, the multikinase inhibitor is selected from the group consisting of nintedanib {(3Z)-3-{[(4-{methyl[(4-methylpiperazine-1-yl)acetyl]amino}phenyl)amino](phenyl)methylidene}-2-oxo-2,3-dihydro-1H-indole-6-carboxylate methyl}, its free base, hydrate, solvate, or pharmaceutically acceptable salt. In some cases, the multikinase inhibitor is nintedanib free base or nintedanib esylate (ethanesulfonate).

[0014] In all of the specified embodiments, the multikinase inhibitor is administered to the affected eye in the form of a topical ophthalmic preparation, a sustained-release semi-solid preparation, a sustained-release solid preparation, or an intraocular implant. In some cases, nintedanib is administered to the affected eye in the form of a topical ophthalmic preparation. In some cases, the topical ophthalmic preparation is a solution, suspension, or emulsion. In all of the specified embodiments, the concentration of nintedanib in the topical ophthalmic preparation is 0.001% to 10% by weight of the total amount of the preparation.

[0015] In all predetermined embodiments, the topical ophthalmic formulation further contains one or more pharmaceutically acceptable additives selected from stabilizers, surfactants, polymer-based carriers, gelling agents, organic auxiliary solvents, pH adjusting components, osmotic pressure adjusting components, and preservatives. Optionally, the sustained-release semi-solid formulation, sustained-release solid formulation, or intraocular implant is inserted into the affected eye. Optionally, the sustained-release semi-solid formulation, sustained-release solid formulation, or intraocular implant contains nintedanib and pharmaceutically acceptable additives. Optionally, the amount of nintedanib in the sustained-release semi-solid formulation, sustained-release solid formulation, or intraocular implant is 1 μg to 100 mg. Optionally, the sustained-release semi-solid formulation, sustained-release solid formulation, or intraocular implant contains nintedanib and a biodegradable polymer selected from polylactic acid (PLA), polyglycolic acid (PLGA), and copolymers of polylactic acid and polyglycolic acid.

[0016] The term "one or more" as used in this application includes the fact that the number designated as "one or more" is at least one, and more appropriately, 1, 2, 3, 4, 5, 10, 20, 50, 100, 500, etc.

[0017] The term "subject" refers to an animal, a human, or one or more cells derived from an animal or a human. The subject is preferably human. The subject may also include non-human primates. Human subjects are sometimes referred to as patients.

[0018] Unless otherwise defined, all scientific and technical terms used in this application have the same meaning as those generally understood by those skilled in the art to which the present invention pertains. While this application describes the methods and materials used in the present invention, other suitable methods and materials known in the art may also be used. These materials, methods, and specific examples are illustrative and not limiting. All publications, patent applications, patents, sequences, database entries, and other reference materials referenced herein are incorporated herein by their entirety. In case of any inconsistency, the specification, including definitions, shall prevail.

[0019] Other features and advantages of the present invention will be readily apparent from the following detailed description and drawings, as well as from the claims.

[0020] This patent or application document shall be accompanied by at least one color drawing. A copy of this patent or the published patent application, including the color drawing, will be issued by the Japan Patent Office after the application and payment of the required fees. [Brief explanation of the drawing]

[0021] [Figure 1A] Figures 1A and 1B are graphs demonstrating the anti-angiogenic effect of nintedanib in a rabbit corneal suture model. Figure 1A shows the results on day 12. It shows the area of ​​corneal neovascularization in each treatment group. Group 1: Positive control 0.05% sunitinib TID; Group 2: 0.2% nintedanib BID; Group 3: 0.2% nintedanib TID; Group 4: 0.05% nintedanib BID; Group 5: 0.05% nintedanib TID; Group 6: Vessel control TID. The significance level of the t-test comparing each group to the vehicle-administered group is indicated by an asterisk. [Figure 1B] Figures 1A and 1B are graphs demonstrating the anti-angiogenic effect of nintedanib in a rabbit corneal suture model. Figure 1B shows the results on day 14. The area of ​​corneal neovascularization in each treatment group is shown. Group 1: Positive control 0.05% sunitinib TID; Group 2: 0.2% nintedanib BID; Group 3: 0.2% nintedanib TID; Group 4: 0.05% nintedanib BID; Group 5: 0.05% nintedanib TID; Group 6: Vessel control TID. The significance level of the t-test comparing each group to the vehicle-administered group is indicated by an asterisk. [Figure 2A] Figure 2A is a graph demonstrating the effect of nintedanib (CBT-001) in a mouse model of human pterygium. 0.2% nintedanib reduced the area of ​​pterygium lesions on the mouse cornea. The pterygium lesion area on days 14 and 17 was significantly smaller than the baseline level on day 7. In contrast, the pterygium area in the eyes of the saline control group increased over 10 days, and was significantly larger on days 14 and 17 than on day 7. [Figure 2B]Figure 2B is a graph demonstrating the effect of nintedanib (CBT-001) in a human pterygium mouse model. 0.2% nintedanib reduced neovascularization on the cornea, and the reduction was significant when comparing the level on day 17 with the baseline on day 7. In the eyes of the saline control group, neovascularization was slightly enhanced, but there was no statistically significant difference from the baseline. [Figure 3A] Figure 3A is a graph demonstrating the effect of sunitinib (CBT-003) in a pterygium mouse model. 0.05% sunitinib showed a tendency to reduce the pterygium lesion area, but the differences at each time point from the baseline on day 7 were not statistically significant. The pterygium area in the eyes of the saline control group increased over 10 days and was significantly larger on days 14 and 17 than on day 7. [Figure 3B] Figure 3B is a graph demonstrating the effect of sunitinib (CBT-003) in a pterygium mouse model. 0.05% sunitinib reduced neovascularization on the cornea, and the reduction was significant when comparing the levels on days 14 and 17 with the baseline on day 7. In the eyes of the saline control group, neovascularization did not change significantly. [Figure 4A] Figure 4A is a graph demonstrating the effect of mitomycin (CBT-002) in a pterygium mouse model. 0.002% mitomycin showed a tendency to reduce the pterygium lesion area, but the differences at each time point from the baseline on day 7 were not statistically significant. The pterygium area in the eyes of the saline control group increased, and was significantly larger on days 10, 14, and 17 than on day 7. [Figure 4B] Figure 4B is a graph demonstrating the effect of mitomycin (CBT-002) in a pterygium mouse model. 0.002% mitomycin reduced neovascularization on the cornea, and the reduction was significant when comparing the level on day 17 with the baseline on day 7. In the eyes of the saline control group, neovascularization did not change significantly. [Figure 5A]Figure 5A is a graph demonstrating the effect of combining nintedanib (CBT-001) and mitomycin (CBT-002) in a pterygium mouse model. The pterygium area in the eyes of the saline control group increased, and was significantly larger on days 14 and 17 than on day 7, but the area did not increase significantly in the combination therapy group. [Figure 5B] Figure 5B is a graph demonstrating the effect of combining nintedanib (CBT-001) and mitomycin (CBT-002) in a pterygium mouse model. Neovascularization area did not change significantly in either the saline administration group or the combination therapy group. [Modes for carrying out the invention]

[0022] Pterygium is an ocular surface disease characterized by fibrovascular proliferation that extends from the nasal or temporal conjunctiva, across the limbus, and into the cornea. Patients with pterygium often experience eye discomfort and redness, and if the lesion invades the visual axis, there is a risk of causing visual impairment. The incidence of pterygium is clearly associated with lifetime sun exposure and other risk factors such as age, male gender, and rural living, and protective effects have been shown by wearing glasses and hats. Furthermore, pterygium is more prevalent in certain occupational groups, such as welders, manual laborers, and outdoor workers, reflecting the important role of ultraviolet (UV) exposure in the pathogenesis of this disease.

[0023] Pterygium is often considered a benign tumor because, despite its invasive growth and recurrence tendencies, it does not metastasize. Current understanding of the pathogenesis of pterygium suggests that multiple processes are involved, which can be divided into genetic factors, environmental triggers (UV radiation, viral infections), and factors that sustain its growth (cytokines, growth factors, and matrix proteases). Of these, chronic UV exposure is the sole and greatest contributing factor to the pathogenesis of pterygium. The relationship between UV exposure and pterygium is well supported by epidemiological studies, as well as its association with other UV-related conditions such as photoaging, cataracts, climatic guttate keratopathy, squamous cell carcinoma, and basal cell carcinoma. UV-activated molecular mechanisms, such as oxidative stress and growth factor receptor (GFR) signaling, lead to the synthesis and secretion of effector molecules that sustain pterygium growth, such as cytokines, growth factors, and matrix proteases. UV radiation is a well-known oxidative stress inducer and also a contributing factor to photoaging. Ultraviolet-induced oxidative stress mediates the activation of the epidermal growth factor receptor (EGFR), which in turn mediates downstream signaling via the mitogen-activated protein kinase pathway.

[0024] Currently, there are no approved drug therapies for treating pterygium. Conjunctival autografting after pterygium resection remains the definitive treatment method for both primary and recurrent pterygium. While many of these lesions can be easily removed, initially satisfying both the surgeon and the patient, pterygium recurrence can occur. To reduce the recurrence rate, antimetabolites such as 5-FU and MMC are used concurrently with or after pterygium surgery (Almond et al., Pterygium: Techniques and Technologies for Surgical Success. Hovanesian JA. Ed. SLACK Incorporated. 2012; pp 55-63).

[0025] Pterygium is a multifactorial disease, and several growth factors, such as VEGF and PDGF, may be involved in its pathogenesis. However, no drugs have been developed to combat these growth factors and treat this disease. The anti-VEGF antibodies bevacizumab and ranibizumab are undergoing clinical trials in pterygium patients worldwide, but the results are very mixed, and it remains unclear whether such antibody-based treatments are effective. A few trials have reported that bevacizumab was able to inhibit pterygium growth, but the majority have reported negative results. To date, there are no reports of small-molecule anti-angiogenic drugs being clinically tested for pterygium. Although there are only two published studies, a human pterygium mouse model has recently been developed (Lee et al. Graefes Arch Clin Exp Ophthalmol. 2014;252(4):609-18; Cox et al. Ophthalmology. 2010;117(9):1782-91). However, anti-angiogenic drugs have not been tested in this model to date. As described below, the inventors are the first to demonstrate that a multi-kinase inhibitor with anti-angiogenic activity effectively inhibits and / or stabilizes pterygium growth and reduces the size of pterygium tissue lesions in a mouse model. Furthermore, the inventors demonstrate that similar results can be obtained with antimetabolites. This disclosure is based in part on these novel findings. Accordingly, this disclosure provides compositions and methods for administering a multi-kinase inhibitor, antimetabolite, or a combination of a multi-kinase inhibitor and antimetabolite to treat pterygium by stabilizing and inducing regression of the pterygium.

[0026] As used in this application, the term "pterygium regression" means a decrease or reduction in the size of the pterygium in the affected eye. For example, the term "pterygium regression" means a decrease or reduction of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100% of the size of the pterygium in the affected eye.

[0027] As used in this application, the terms "conjunctival congestion" or "conjunctivitis" refer to a condition where there is excessive blood flow to the white of the eye (sclera), causing the eye to become red. The term "reduce congestion" refers to a reduction in the red area and / or an increase in the white area of ​​the affected eye. The reduction in the red area and / or the increase in the white area can be confirmed or measured by methods well known to those skilled in the art, including visual evaluation by a specialist.

[0028] The terms "stabilizing the pterygium" or "stabilizing the size of the pterygium" as used in this application mean maintaining the size of the pterygium in the affected eye.

[0029] The term "pterygium recurrence" as used in this application refers to the reappearance of a pterygium in the eye after removal of the primary pterygium (e.g., surgical removal).

[0030] As used in this application, the terms “therapeutic effectiveness” and “effective dose” mean the amount of drug effective in producing the target pharmacological, therapeutic, or preventive outcome. A pharmacological effective dose is one that results in improvement of one or more symptoms of a disorder, prevents the progression of the disorder, causes regression of the disorder, or prevents the disorder. For example, with respect to the induction of pterygium regression, a therapeutic effective dose means the amount of drug that reduces the size of the pterygium by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.

[0031] The effective therapeutic dose in the method described herein can be determined by the treating physician. For example, a physician may initiate treatment with the manufacturer's recommended dose for a multi-kinase inhibitor or antimetabolite and adjust it based on the physician's observation of the therapeutic effect. Detailed guidance is provided in this specification and examples. Furthermore, clinical trials may be conducted to determine an effective dose that produces a statistically significant therapeutic effect when treating a patient population.

[0032] The terms "or a combination thereof" or "in combination with" refer to all forms of administration in which a first drug is provided together with a second drug, such as a second inhibitory nucleic acid molecule or chemotherapeutic agent, and these two drugs are administered simultaneously or sequentially in any order. When two or more drugs are administered in combination with each other, it is not necessary to administer them simultaneously or as identical formulations. Drugs administered in combination with each other exhibit or possess simultaneous biological activity in the body of the target to which they are delivered. Whether or not a drug is present in the target's body can be easily determined by empirical monitoring or by calculation using the known pharmacokinetic properties of the drug.

[0033] The terms “treatment,” “therapeutic,” and “to treat” are generally used in this application to mean obtaining a desired pharmacological and / or physiological effect. The effect may be a preventive effect, meaning completely or partially preventing the disease or its symptoms, and / or a therapeutic effect, meaning the partial or complete stabilization or cure of the disease and / or adverse effects caused by the disease. The term “treatment” encompasses all treatments of diseases in mammals, especially humans, and includes (a) preventing the onset of the disease and / or symptoms in subjects who appear to have a predisposition to the disease or symptoms but have not yet been diagnosed as having the disease; (b) preventing the onset of the disease and / or symptoms, i.e., inhibiting its development; or (c) alleviating the symptoms of the disease, i.e., reducing the disease and / or symptoms. Those who require treatment include those who are already ill (e.g., cancer patients, infected persons, etc.) and those for whom prevention is desired (e.g., those who are prone to cancer, those who are susceptible to infection, those suspected of having cancer, those suspected of having infection, etc.).

[0034] As used in this application, the term "multi-kinase inhibitor" (MKI) means a pharmaceutical compound (e.g., a small molecule) that reduces or inhibits the expression or activity of two or more kinases, including, for example, intracellular and / or cell surface protein kinases.

[0035] As used in this application, "low molecular weight" means a compound with a molecular weight of less than 2,000 daltons, more preferably 200 to 1,000 daltons, and even more preferably 300 to 700 daltons. These low molecular weights are preferably organic molecules. In certain embodiments, the "low molecular weight" does not include peptides or nucleic acid molecules.

[0036] The exemplary multikinase inhibitors used in the methods described herein exhibit a specific kinase inhibition profile. For example, the multikinase inhibitors used in the methods described herein exhibit IC50 for VEGFR(1,2,3). 50 <200nM, and IC for PDGFR(α,β) 50 <200nM, and IC relative to FGFR(1, 2, 3) 50It can have a kinase inhibition profile with an IC50 of <1 μM.

[0037] Examples of multikinase inhibitors used in the method described herein include, for example, afatinib, amvatinib, axitinib, cabozantinib, canertinib, cedilanib, ceritinib, clenolanib, crizotinib, dabrafenib, dacomitinib, dasatinib, erlotinib, foretinib, gefitinib, golbatinib, ibrutinib, and icoti. Examples include nib, idelalisib, imatinib, lapatinib, lenvatinib, neratinib, nilotinib, nintedanib, palbociclib, pazopanib, ponatinib, quizartinib, regorafenib, ruxolitinib, sorafenib, sunitinib, tanzutinib, civantinib, cibozanib, trametinib, vandetanib, batalanib, and vemurafenib.

[0038] Nintedanib {(3Z)-3-{[(4-{methyl[(4-methylpiperazine-1-yl)acetyl]amino}phenyl)amino](phenyl)methylidene}-2-oxo-2,3-dihydro-1H-indole-6-carboxylate methyl} is an example of a multi-kinase inhibitor as described in this application. Nintedanib primarily inhibits receptor tyrosine kinases such as vascular endothelial growth factor receptor (VEGFR1-3), platelet-derived growth factor receptor (PDGFRα and β), and fibroblast growth factor receptor (FGFR1-4) (see Table 1 below), exhibiting a unique kinase inhibition profile.

[0039] [Table 1]

[0040] As used in this application, the term “antometabolic antagonist” means a pharmaceutical compound that inhibits the utilization of metabolites and therefore reduces, interferes with, or inhibits the proliferation of rapidly dividing cells. For example, the antimetabolic antagonists of this disclosure can inhibit DNA replication by various mechanisms that result in the reduction, interference with, or inhibition of cell division. Examples of antimetabolic antagonists used in the methods described herein include, for example, purine and pyrimidine analogs (e.g., 5-fluorouracil (5-FU)), antibiotics (e.g., mitomycin C (MMC)), and antifolate compounds (e.g., methotrexate). Examples of antimetabolic antagonists used in the methods described herein include, for example, mitomycin C, 5-fluorouracil, phloxuridine, cytarabine, 6-azauracil, azathioprine, methotrexate, mycophenolate mofetil, and thiotepa.

[0041] Mitomycin C (MMC) is an example of an antimetabolite as described in this application. MMC is an antitumor antibiotic that is activated by reduction and becomes a potent alkylating agent. Under hypoxic conditions, it is most effective in actively dividing cells because it interferes with DNA replication by crosslinking DNA at the N2 position of guanine. Under aerobic conditions, it generates toxic oxygen radicals that can nonspecifically interfere with RNA and protein synthesis. MMC is used intravenously as an antitumor agent, particularly for tumors of the gastrointestinal tract, pancreas, lung, and breast. It is also used intravesically for bladder cancer.

[0042] 5-FU is an example of an antimetabolite as described in this application. 5-FU is a fluoropyrimidine, and its primary antimetabolite effect is the inhibition of thymidylate synthase, which is thought to result in a deficiency of intracellular thymidine for DNA production. In addition, 5-FU has other effects resulting from the inhibition of other enzymes or the incorporation of their metabolites into RNA.

[0043] Multi-kinase inhibitors and antimetabolites are well known to those skilled in the art and are widely used in the treatment of cancer.

[0044] In the early stages of pterygium, pinguecula, or pseudopterygium, patients experience eye discomfort, redness, irritation, blurred vision, foreign body sensation, and pain. The compositions and methods described herein are useful in treating such patients, with the goal of reducing redness and symptoms. For example, this disclosure provides compositions and therapeutic methods using nintedanib, a multi-kinase inhibitor, in an appropriate ophthalmic dosage form to reduce redness and other symptoms.

[0045] In the later stages of pterygium, when fibrovascular proliferation extends from the conjunctiva beyond the limbus to the center of the cornea and invades the visual axis, the generally selected treatment method has been pterygium resection by scleral exposure followed by conjunctival autograft or amniotic membrane grafting. The compositions and methods described herein are also useful for treating such patients. Although the risk of recurrence has decreased significantly with advances in surgical techniques and adjuvant therapies, recurrence remains a significant problem for surgeons and patients. For the purpose of preventing pterygium recurrence, this disclosure provides compositions and treatment methods for using nintedanib in an appropriate ophthalmic dosage form to reduce pterygium recurrence.

[0046] As shown in the rabbit suture model study described in Example 1, the compositions and methods described in this application are also useful for treating hyperemia and alleviating associated symptoms. In this example, nintedanib was highly effective in inhibiting neovascularization induced by sutures in the cornea. As shown in Table 2 below, the percentage change in neovascular area was substantially reduced in the eyes administered with nintedanib compared to the base-administered group. The effect of nintedanib depends on the dose concentration and administration frequency regimen. Interestingly, nintedanib showed a clearly superior trend to sunitinib in terms of reducing neovascular area. Although the targets of these two kinase inhibitors substantially overlap, different effects were observed in this rabbit model. Another advantage of nintedanib is that it inhibits fewer targets than sunitinib, resulting in a better safety limit and the ability to increase to higher doses. In fact, as shown in Example 1, one rabbit in the sunitinib-treated group exhibited a potential toxicity in the form of lens abnormalities, including partial opacity of the lens, during the administration period, while this was not observed at all in the nintedanib-treated group. Furthermore, in an in vivo corneal suture rabbit model study in which sunitinib was administered for 7 days after sutures were placed in the cornea of ​​rabbits, an anterior chamber cell response was observed on day 1 (Perez-Santonja JJ et al, Am J Ophthalmol. 2010;150(4):519-528). Yellowish discoloration of the iris surface, indicating sunitinib deposition, was observed throughout the entire administration period. Sunitinib is deposited in the inferior quadrant of the iris, starting from the pupillary margin, and spreads to some extent between the pupil and the anterior chamber angle, potentially causing long-term toxic effects. Furthermore, at concentrations of >3.3 μg / mL, significant toxicity of sunitinib was observed after 24-hour incubation in human corneal epithelial cells (Bayyoud T et al., Current Eye Research, 39(2):149-154, 2014).

[0047] [Table 2]

[0048] Both nintedanib and sunitinib inhibit major VEGFR and PDGFR families, but several targets do not overlap (see Table 1). However, nintedanib appears to be more effective and safer than sunitinib due to its unique combination of kinase targets, and it appears to be one of the most potent multi-kinase inhibitors for reducing corneal neovascularization. Specifically, the unique inhibitory profiles of nintedanib, as expressed by its maximum in vitro IC50, against the following intracellular and / or cell surface protein kinases are: VEGFR(1, 2, 3) (IC50 < 50 nM), PDGFR(α, β) (IC50 < 100 nM), FGFR(1, 2, 3) (IC50 < 150 nM), FGFR4 (IC50 < 1000 nM), FLT3 (IC50 < 50 nM), Lck (IC50 < 50 nM), Lyn (IC50 < 200 nM), and Src (IC50 < 200 nM). The three targets of FGFR4, Lyn, and Src are not inhibited by sunitinib or other common kinase inhibitors, and these targets allow nintedanib to be distinguished from sunitinib and others. Furthermore, nintedanib has substantially higher efficacy against FGFR1-3 than sunitinib, which is likely one of the reasons why nintedanib shows superior results in the corneal suture rabbit model.

[0049] In the mid-stage of pterygium, fibrovascular proliferation extends from the conjunctiva to the limbus and cornea, and the compositions and methods described herein are also useful in treating such patients. In the mid-stage of pterygium, the goal is to stabilize pterygium progression, delay or avoid surgery to remove the pterygium, or induce pterygium regression from the visual axis / corneal center. To achieve this goal, the disclosure provides compositions and therapeutic methods using multi-kinase inhibitors, antimetabolites, or combinations thereof in appropriate ophthalmic dosage forms.

[0050] An example of a composition and method for treating an affected eye using a multi-kinase inhibitor to stabilize pterygium and induce pterygium regression is described in Example 2 below, demonstrating the effect of such a composition on the proliferation of human pterygium cells on the cornea of ​​immunodeficient mice. In this study, nintedanib and sunitinib inhibited the growth of human pterygium on the mouse cornea, and nintedanib significantly reduced the size of the pterygium. As shown in Table 3 below, pterygium cells proliferated throughout the treatment period up to day 17, but in the group treated with nintedanib, sunitinib, MMC, or a combination of nintedanib and MMC, pterygium cells did not proliferate or regressed in the case of nintedanib. Therefore, our novel insights from the mouse model demonstrate that multi-kinase inhibitors such as nintedanib or sunitinib can be used to inhibit pterygium growth or even induce pterygium tissue regression. As an example, the target kinase profile of the multi-kinase inhibitor used in the composition and method described herein is the following kinases in specific in vitro IC 50 It can be targeted with VEGFR(1,2,3)(IC 50 <200nM), PDGFR(α,β)(IC 50 <200nM), FGFR(1,2,3)(IC 50 (<1 μM).

[0051] [Table 3]

[0052] The compositions and methods described herein are also useful for treating patients using antimetabolites, which are inhibitors of epithelial and fibroblast proliferation, to stabilize pterygium and induce pterygium regression (see Example 2). As shown in Table 3 above, mitomycin C (MMC) was able to prevent the proliferation of human pterygium cells and showed a tendency to reduce the size of pterygium tissue on the cornea.

[0053] Considering the multifactorial nature of pterygium, it appears that combination therapy with multiple drugs is necessary to achieve optimal efficacy. As shown in Table 3, pterygium cells on the cornea treated with nintedanib and MMC did not show significant proliferation, but they proliferated significantly in saline control.

[0054] Formulations and administration regimens The method described in this application includes the manufacture and use of a pharmaceutical composition containing a compound identified as an active ingredient by the method described in this application. Such a pharmaceutical composition itself is also included in this application.

[0055] Pharmaceutical compositions generally contain pharmaceutically acceptable additives. As used in this application, the terms "pharmaceutically acceptable additive" or "pharmaceutically acceptable carrier" include physiological saline, solvents, dispersions, coatings, antibacterial agents, antifungal agents, isotonic agents, and absorption retarders that are suitable for drug administration.

[0056] Appropriate methods for formulating pharmaceutical compositions are well known in this field; see, for example, Remington: The Science and Practice of Pharmacy, 21st ed., 2005, and the series of textbooks in Drugs and the Pharmaceutical Sciences: A Series of Textbooks and Monographs (Dekker, NY). For example, solutions, suspensions, or emulsions used for ophthalmic applications may contain the following components: sterile diluents such as sterile water for injection, physiological saline, non-volatile oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents; antioxidants; chelating agents; buffers such as acetates, citrates, or phosphates; and tonicity adjusters such as sodium chloride or glucose. The pH can be adjusted with an acid or base such as hydrochloric acid or sodium hydroxide.

[0057] Suitable pharmaceutical compositions for injection include sterile aqueous solutions (if water-soluble) or dispersions, and sterile powders for the immediate preparation of sterile injection solutions or dispersions. These compositions must be stable under manufacturing and storage conditions and must be protected from contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol) and suitable mixtures thereof. Appropriate fluidity can be maintained by using coating agents such as lecithin, maintaining the required particle size in the case of dispersions, or by using surfactants. Microbial activity can be prevented by various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc.). In many cases, it is preferable to include isotonic agents (e.g., sugars, polyalcohols such as mannitol and sorbitol, and sodium chloride) in the composition. Long-term absorption of the injection composition can be achieved by including substances that slow absorption (e.g., aluminum monostearate and gelatin) in the composition.

[0058] Sterile injection solutions can be prepared by compounding the required amount of the active compound with one or a combination of the above components in a suitable solvent, and then sterilizing by filtration. Generally, dispersions are prepared by compounding the active compound with a sterile base containing a basic dispersion medium and other necessary components selected from the above components. In the case of sterile powders for the preparation of sterile injection solutions, preferred manufacturing methods are vacuum drying and freeze-drying, from which powders of the active ingredient and other desired components can be obtained from a pre-sterilized and filtered solution.

[0059] In one embodiment, the therapeutic compound is manufactured using a carrier that protects the therapeutic compound from rapid excretion from the body (e.g., a controlled-release formulation including an implant and a microcapsule delivery system). Biodegradable and biocompatible polymers such as ethylene vinyl acetate, polyacid anhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Such formulations can be manufactured using standard techniques or are available commercially.

[0060] The pharmaceutical composition can be contained in a container, pack, or dispenser with an instruction sheet for administration attached.

[0061] Compositions and formulations containing multi-kinase inhibitors, antimetabolites, or combinations of multi-kinase inhibitors and antimetabolites as described herein may be administered topically, inserted as semi-solid formulations or solid implants, or administered by any other suitable method known in the art. While the agents disclosed herein may be used alone for treatment, it is preferable to administer the agents as pharmaceutical formulations mixed with appropriate pharmaceutical excipients, diluents, or carriers selected, for example, in view of the intended route of administration and standard pharmaceutical practice. The pharmaceutical formulations contain at least one active compound together with pharmaceutically acceptable excipients, diluents, and / or carriers.

[0062] The composition or preparation may be administered once, twice, three times, four times, or more times per day. During the maintenance phase of treatment with the therapeutic agent, the frequency may be reduced, for example, once every two or three days instead of daily or twice daily. The dosage and frequency of administration may be adjusted at the discretion of the treating physician, taking into consideration, for example, the clinical signs, pathological signs, and clinical and subclinical symptoms of the disease being treated by the method of this application, as well as the patient's clinical history.

[0063] Naturally, the amount of the drug disclosed herein required for therapeutic use will vary depending on the route of administration, the type of condition requiring treatment, and the patient's age, weight, and health condition, and will ultimately be left to the discretion of the attending physician. The compositions generally contain an effective amount of the active ingredient as a single agent or combination. Provisional doses can be determined according to animal studies, and the dose can be increased or decreased to suit human administration in accordance with generally accepted practices in the art.

[0064] The treatment period, i.e., the number of days, is easily determined by the physician treating the patient, but the treatment period can range from approximately 1 day to approximately 365 days. The therapeutic effect provided by the method of this invention can be monitored during the treatment period, and it is possible to determine whether the treatment is successful or whether additional (or modified) treatment is necessary.

[0065] For example, the dosage, toxicity, and therapeutic effect of therapeutic compounds can be determined in cell cultures or experimental animals using standard pharmaceutical methods to determine the LD50 (the dose at which 50% of the population dies) and ED50 (the dose at which 50% of the population shows a therapeutic effect). The dosage and formulation of multi-kinase inhibitors and antimetabolites, as well as their individual dose strengths in combination therapy, can be easily determined by those skilled in the art, and can be obtained, for example, from animal models and clinical trials reported in the literature as means of determining dosage, safety, and efficacy according to standard methods known in the art. The exact prescription, route of administration, and dosage can be selected by individual physicians taking into account the patient's health condition.

[0066] For example, the dosage strength of multikinase inhibitors is approximately 0.001 to 100.0 mg, 0.01 to 90 mg, 0.1 mg to 75 mg, 0.25 to 50 mg, 0.5 to 25 mg, 0.75 to 20 mg, 1.0 to 15 mg, 1.25 to 10 mg, 1.5 to 5.0 mg, 1.75 to 2.5 mg, and 0. Examples include 0.001 mg, 0.01 mg, 0.1 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1.0 mg, 1.25 mg, 1.5 mg, 1.75 mg, 2.0 mg, 2.5 mg, 5.0 mg, 10.0 mg, 15.0 mg, 25.0 mg, 30.0 mg, 40.0 mg, 50.0 mg, 60.0 mg, 75.0 mg, or 100.0 mg. For example, possible dosages of nintedanib include 0.1 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1.0 mg, 1.25 mg, 1.5 mg, 1.75 mg, 2.0 mg, 2.5 mg, 5.0 mg, 10.0 mg, 15.0 mg, 25.0 mg, 30.0 mg, 40.0 mg, 50.0 mg, 60.0 mg, 75.0 mg, or 100.0 mg.

[0067] The compositions used in the present invention may contain a multikinase inhibitor at a concentration of 0.001% to 10% of the total composition by weight or volume. For example, an aqueous composition may contain nintedanib at concentrations of 0.001%, 0.01%, 0.1%, 0.5%, 1.0%, 1.5%, 2.0%, 5.0%, or up to 10%.

[0068] For example, the dosage strength of antimetabolites is approximately 0.001 to 100.0 mg, 0.01 to 90 mg, 0.1 mg to 75 mg, 0.25 to 50.0 mg, 0.5 to 25 mg, 0.75 to 20 mg, 1.0 to 15 mg, 1.25 to 10 mg, 1.5 to 5.0 mg, 1.75 to 2.5 mg, for example 0 Examples include 0.001 mg, 0.01 mg, 0.1 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1.0 mg, 1.25 mg, 1.5 mg, 1.75 mg, 2.0 mg, 2.5 mg, 5.0 mg, 10.0 mg, 15.0 mg, 25.0 mg, 30.0 mg, 40.0 mg, 50.0 mg, 60.0 mg, 75.0 mg, or 100.0 mg. For example, possible dosage intensities for MMC include 0.1 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1.0 mg, 1.25 mg, 1.5 mg, 1.75 mg, 2.0 mg, 2.5 mg, 5.0 mg, 10.0 mg, 15.0 mg, 25.0 mg, 30.0 mg, 40.0 mg, 50.0 mg, 60.0 mg, 75.0 mg, or 100.0 mg.

[0069] The compositions used in the present invention may contain an antimetabolite at a concentration of 0.001% to 10% of the total composition by weight or volume. For example, an aqueous composition may contain up to 0.001%, 0.01%, 0.1%, 0.5%, 1.0%, 1.5%, 2.0%, 5.0%, or 10% of MMC.

[0070] As will be obvious to those skilled in the art, aqueous solutions can be administered to the eyes as "droplets" or multiple droplets (e.g., multi-kinase inhibitor solutions, antimetabolite solutions, or combinations thereof) from a dropper, pipette, or other dedicated sterile instrument. Such droplets generally have a volume of up to 50 microliters, but may be smaller, for example, less than 10 microliters. [Examples]

[0071] The present invention will be described in more detail below with reference to examples, but the following examples are not intended to limit the scope of the present invention as described in the claims.

[0072] [Example 1]: Rabbit corneal suture model of neovascularization and hyperemia A rabbit corneal suture model has been established to evaluate the effects of pharmaceuticals on corneal neovascularization (Ko et al. Cornea. 2013; 32(5): 689-695; Perez-Santonja et al. Am J Ophthalmol. 2010; 150(4): 519-528). Nintedanib was tested for its anti-neovascular activity in this model.

[0073] Topical ophthalmic preparations Topical compositions containing 0.2% or 0.05% nintedanib were prepared in phosphate buffer (pH 7.4) supplemented with 10% 2-hydroxypropyl-β-cyclodextrin. A composition containing 0.05% sunitinib on the same base was also prepared and used as a positive control.

[0074] Animals and treatment procedures The study was conducted using 36 female New Zealand White rabbits. In summary, on day 1, sutures were placed in five locations in the corneal epithelium of each animal's right eye to induce neovascularization. The drug was administered to both eyes of the animals as shown in Table 4.

[0075] [Table 4]

[0076] During the experiment, the animals were closely observed for various eye signs and general health conditions, including body weight. Eye images were taken for analysis on days 7, 10, 12, 14, 21, and 28.

[0077] Data Analysis Eye images were analyzed using NIH ImageJ(R) software. Each image was opened in ImageJ(R), scales were set on the photograph using the ruler, and neovascularization areas on the cornea near sutures were selected using the selection tool. The area (mm²) was measured using the software's measurement tool.2 The values ​​were calculated, recorded in Excel, images were captured, and saved. A two-tailed t-test was used to determine whether there was a significant difference between each pair of groups. The results were plotted as histograms of the mean and standard deviation for easy comparison.

[0078] Results and Discussion The results of this study are summarized in Figures 1A and 1B and Table 5 below. These results demonstrated that nintedanib had a significant inhibitory effect on suture-induced neovascularization in rabbit corneas. High-dose 0.2% nintedanib showed improved efficacy compared to 0.05% nintedanib, and the high-frequency dosing regimen (TID) showed improved efficacy compared to BID. Surprisingly, nintedanib showed a clear superior trend to the positive control sunitinib in terms of reducing neovascular area in this model.

[0079] In a rabbit suture model, sunitinib has been shown to inhibit angiogenesis more effectively than the anti-VEGF antibody bevacizumab (Ko et al. Cornea. 2013;32(5):689-695; Perez-Santonja et al. Am J Ophthalmol. 2010;150(4):519-528). These observations suggest that small molecule kinase inhibitors targeting multiple receptor tyrosine kinase pathways may have advantages over antibody drugs targeting highly selective pathways. In this study, nintedanib was also found to inhibit neovascularization very effectively in the suture model. Completely unexpectedly, the inventors discovered that kinase inhibitors may have different efficacy even if their kinase targets substantially overlap. Both nintedanib and sunitinib inhibit major VEGFR family members, but several of their targets do not overlap. As shown in Table 1, the target profile of nintedanib differs from that of sunitinib and several other MKIs that are considered very similar in the research community. The unique combination of kinase targets of nintedanib appears to provide at least two remarkable advantages: 1) Nintedanib is a highly effective inhibitor of neovascularization, more effective than sunitinib, as demonstrated in the rabbit suture model; 2) The number of targeted kinases is smaller than that of sunitinib, resulting in better safety limits and the ability to increase the dose. For these reasons, it is presumed that nintedanib shows improved efficacy and a better safety profile compared to sunitinib.

[0080] [Table 5]

[0081] The rationale for the inventors' claims is supported by previous research findings in cancer research, an area in which MKIs are widely used. In cancer, several MKIs with substantially overlapping targets may still exhibit very different efficacy in patients. For example, while numerous small molecule MKIs with overlapping targets have been tested in patients with non-small cell lung cancer (NSCLC), interestingly, only nintedanib has shown efficacy and has been endorsed in combination therapy with other drugs (Hall RD et al. Transl Lung Cancer Res. 2015;4(5), 515-23). ​​Such observations demonstrate that the target profiles of kinase inhibitors can exert dramatic efficacy in specific indications.

[0082] The inventors' novel insights can be summarized as follows: First, generally speaking, small molecule MKIs with specific target profiles are superior to antibody drugs for the treatment of abnormal corneal neovascularization. This had been implied in previous studies, but only through comparisons of the bevacizumab and sunitinib pair. Our current testing of nintedanib significantly increases the weight of this theory. Second, our novel idea is that while each MKI has a unique target profile, not all MKIs are equally safe and effective in treating pterygium symptoms. Without being bound by theory, we believe that nintedanib offers a unique profile that enables one of the most effective and stable treatments for pterygium.

[0083] In short, nintedanib, with its unique target profile, could be a more effective and safer treatment for pterygium. Nintedanib inhibits FGFR1-3 more effectively than sunitinib. It also inhibits FGFR4, Lyn, and Src, which are not targets of sunitinib (see Table 1 in the Background Technology section). Furthermore, sunitinib is known to attack more kinases than compounds in its class (Kumar et al. Br J Cancer. 2009;101(10):1717-23), and inhibits several other kinases not listed in Table 1, so nintedanib is expected to have a better safety profile than sunitinib. Some of these kinases, such as the CaMK family, are important for normal cellular function, and inhibition of these could lead to safety issues.

[0084] [Example 2]: Human pterygium mouse model A human pterygium mouse model has been described to evaluate human pterygium growth on the cornea of ​​immunodeficient mice (Lee et al., Graefes Arch Clin Exp Ophthalmol. 2014;252(4):609-18). In this study, we investigated the effects of several pharmaceuticals on pterygium growth. These pharmaceuticals are nintedanib, sunitinib, and mitomycin C.

[0085] Topical ophthalmic preparations The test drug was prepared in phosphate buffer (pH 7.4) supplemented with 10% 2-hydroxypropyl-β-cyclodextrin. Detailed information about the formulation is disclosed in the following section.

[0086] animal Seven-week-old male athymoid nude mice were adapted to pathogen-free conditions in sealed filter-top cages.

[0087] Human pterygium primary cell culture Human pterygium epithelial cells (hPEC) were isolated and cultured from specimens taken after surgical resection. All participants received a comprehensive explanation of the study and then submitted written informed consent. Fresh pterygium specimens were cultured for 3 days in DMEM / F12 medium supplemented with 10% fetal bovine serum, 0.5% dimethyl sulfoxide, and 1% antibiotic / antifungal agent on a surface coated with collagen (rat tail type I collagen). During this period, the cells migrated from the explants. Subsequently, the explants were removed, and the medium was changed to serum-free keratinocyte medium supplemented with 5% BCS and 1% antibiotic / antifungal agent to further promote epithelial cell proliferation.

[0088] Induction of human pterygium For subconjunctival injection, mice were anesthetized by intraperitoneal injection of ketamine (30 mg / kg) and rompun (2.5 mg / kg). On day 0, 1×10 4 hPEC were injected into the nasal subconjunctival space of both eyes to induce pterygium in mice. Seven days later, the mice induced with hPEC were used for the study.

[0089] Treatment The animals were treated as follows. Group 1: Vehicle in the right eye and saline in the left eye; Group 2: 0.2% nintedanib in the right eye and saline in the left eye; Group 3: 0.002% mitomycin in the right eye and saline in the left eye; Group 4: 0.05% sunitinib in the right eye and saline in the left eye; Group 5: A mixture of 0.2% nintedanib and 0.002% mitomycin in the right eye and saline in the left eye.

[0090] On days 7, 10, and 14, subconjunctival injections were performed, and the animals were treated by topical eye drops four times a day on days 8, 9, 11, 12, 13, 15, and 16. Before each injection and on day 17, the eyes were observed using a stereomicroscope and photographed.

[0091] Clinical findings During the experiment, mice were observed daily for signs of clinical toxicity. Their eyes were observed and photographed before injection and on day 17. All findings, including disease, diagnosis, and treatment, were recorded. The body weight of the mice was measured on days 0, 7, 11, 15, and 17.

[0092] Corneal pterygium analysis Image analysis of photographs was performed using ImageJ(R), and the size of the lesion was measured on days 0, 7, 10, 14, and 17. These data were calculated as the ratio of the pterygium to the entire cornea.

[0093] Corneal neovascularization analysis The clinical characteristics of the eyes of all mice were evaluated. The degree of corneal neovascularization (NV) was scored on a scale of 0 to 3, with 0 = no NV, 1 = NV limited to the periphery of the cornea, 2 = NV extending to the pupillary margin, and 3 = NV extending beyond the pupillary margin to the center of the cornea.

[0094] statistical analysis The data was analyzed using SPSS version 18.0 for Windows (SPSS, Chicago, IL) and expressed as mean ± standard deviation.

[0095] Results and Discussion The results are shown in Figures 2A-B, 3A-B, 4A-B, and 5A-B, and further summarized in Table 4 above. In the human pterygium mouse model, administration of 0.2% nintedanib resulted in a reduction of pterygium area on days 14 and 17 compared to the baseline level on day 7 (Figures 2A and 2B). In contrast, the eyes of the control group, which were administered saline, showed an increase in pterygium area on days 14 and 17. Nintedanib also reduced the neovascularization score on the cornea during the administration period, with a significant difference at day 17 compared to the baseline at day 7 (Figures 2A and 2B), while the eyes of the control group showed a slight increase in neovascularization that was not statistically significant (Figures 2A and 2B). In this model, 0.002% mitomycin and 0.05% sunitinib also showed a tendency towards reduction in pterygium area, but did not reach statistical significance at any time point (Figures 3A-3B, 4A-4B). In contrast, the eyes of the control group administered with saline showed an increase in pterygium area at almost all time points, and the increase was almost linear over time. Mitomycin and sunitinib also reduced the neovascularization score, with mitomycin showing a significant decrease on day 17 and sunitinib showing a significant decrease on days 14 and 17 (Figures 3A-3B, 4A-4B). In this case as well, the eyes of the control group did not show a significant change in neovascularization during the administration period. In eyes administered with nintedanib and mitomycin, pterygium area did not increase, but it increased significantly in the saline control group (Figures 5A-5B). The neovascularization score did not show a significant change in this animal group.

[0096] [Example 3]: Formulation Nintedanib ophthalmic solution This pharmaceutical preparation is an isotonic ophthalmic solution prepared with 2-hydroxypropyl-β-cyclodextrin or other similar cyclodextrins in a buffer solution with a pH range of 5.5 to 8.0. Other viscosity modifiers, lubricants, and preservatives may be added to enhance the functionality of the preparation. The composition of this ophthalmic solution is disclosed in Table 6.

[0097] [Table 6] JPEG2026102822000007.jpg91150

[0098] Nintedanib ophthalmic suspension This pharmaceutical preparation is an isotonic ophthalmic suspension prepared with sodium carboxymethylcellulose and a buffer solution with a pH range of 5.5 to 8.0. The particle size of the active pharmaceutical ingredient is reduced to less than 40 microns. Other viscosity modifiers, lubricants, solubilizers, and preservatives may be added to enhance the functionality of the suspension. The composition is disclosed in Table 7.

[0099] [Table 7] JPEG2026102822000009.jpg91150

[0100] Nintedanib ophthalmic emulsion This pharmaceutical preparation is an isotonic ophthalmic emulsion. The active pharmaceutical ingredient is dissolved in a mixed oil phase and an emulsifier, then emulsified and mixed with an aqueous phase with a pH range of 5.5 to 8.0. Other viscosity modifiers, lubricants, solubilizers, and preservatives may be added to enhance the functionality of the emulsion preparation. The composition is disclosed in Table 8.

[0101] [Table 8] JPEG2026102822000011.jpg30150

[0102] Nintedanib sustained-release semi-solid formulation This pharmaceutical preparation is an isotonic sustained-release semi-solid formulation. The active pharmaceutical ingredient is dissolved and / or suspended in a semi-solid medium with a pH range of 5.5 to 8.0. Other viscosity modifiers, lubricants, solubilizers, and preservatives may be added to enhance the functionality of the sustained-release semi-solid formulation. The composition is disclosed in Table 9.

[0103] [Table 9]

[0104] Nintedanib sustained-release implant This pharmaceutical preparation is a solid implant. The active pharmaceutical ingredient (API) is mixed and blended with one or more polymers. The mixture of API and polymers is melted at a predetermined temperature and extruded into a filament of a predetermined diameter. The preparation filament is cut into segments of a predetermined size that can be implanted in ocular tissue. The composition is disclosed in Table 10.

[0105] [Table 10]

[0106] The exemplary compositions used in the methods of the present invention may be modified from existing ophthalmally acceptable compositions.

[0107] Other embodiments The present invention has been described above, including a detailed explanation of the present invention. Naturally, the above description is intended to specifically illustrate the present invention and is not intended to limit the scope of the present invention. The scope of the present invention is defined by the following claims. Other embodiments, advantages and modifications are also included in the following claims.

Claims

1. A method for inducing pterygium regression from the visual axis / corneal center, comprising administering a therapeutically effective amount of (1) a multikinase inhibitor; (2) an antimetabolite that inhibits epithelial cell proliferation and fibroblast proliferation; or (3) a combination thereof to the affected eye of a subject requiring treatment.

2. The method according to claim 1, wherein the size of the pterygium in the affected eye is reduced as a result of administration of the multi-kinase inhibitor, the antimetabolite, or a combination thereof.

3. The method according to claim 1, wherein the pterygium growth rate in the affected eye becomes negative as a result of administration of the multi-kinase inhibitor, the antimetabolite, or a combination thereof.

4. A method for stabilizing a pterygium, comprising administering a therapeutically effective amount of (1) a multikinase inhibitor; (2) an antimetabolite that inhibits epithelial cell proliferation and fibroblast proliferation; or (3) a combination thereof to the affected eye of a subject requiring such treatment.

5. The method according to claim 4, wherein the size of the pterygium in the affected eye is stabilized as a result of administration of the multi-kinase inhibitor, the antimetabolite, or a combination thereof.

6. The method according to claim 4, wherein, as a result of administering the multi-kinase inhibitor, the anti-angiotensin, or a combination thereof, the pterygium growth rate in the affected eye becomes nearly zero.

7. The method according to any one of claims 1 to 6, wherein the multikinase inhibitor reduces the activity of one or more intracellular and / or cell surface protein kinases selected from EGFR, ErbB2, ErbB3, FFFR1, FFFR2, FFFR3, FFFR4, TrkA, NGFR, VEGFR(1,2,3), PDGFR(α,β), TGF-βR(I,II,III), FLT3, Lck, Lyn, Src, c-Kit, c-Fms, Raf-1, B-Raf, RET, and CSF-1R in the pterygium.

8. The multi-kinase inhibitor is IC50 for VEGFR(1,2,3) is <200nM, The IC50 for PDGFR(α,β) is <200 nM, and / or The method according to claim 7, wherein IC50 for FGFR(1,2,3) is <1000 nM.

9. The aforementioned multikinase inhibitors include afatinib, amvatinib, axitinib, cabozantinib, canertinib, cejilanib, ceritinib, clenolanib, crizotinib, dabrafenib, dacomitinib, dasatinib, erlotinib, foretinib, gefitinib, golbatinib, ibrutinib, icotinib, idelalisib, imatinib, lapatinib, and lenvatinib. The method according to any one of claims 1 to 8, selected from the group consisting of bu, neratinib, nilotinib, nintedanib, palbociclib, pazopanib, ponatinib, quizartinib, regorafenib, ruxolitinib, sorafenib, sunitinib, tanzutinib, tivantinib, tivozanib, trametinib, vandetanib, batalanib, and vemurafenib.

10. The method according to any one of claims 1 to 6, wherein the antimetabolite is selected from the group consisting of mitomycin C, 5-fluorouracil, phloxuridine, cytarabine, 6-azauracil, azathioprine, methotrexate, mycophenolate mofetil, and thiotepa.

11. The method according to any one of claims 1 to 10, wherein the multi-kinase inhibitor, the antimetabolite, or a combination thereof is administered to the affected eye in the form of a topical ophthalmic preparation, ointment, gel, sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant.

12. The method according to claim 11, wherein the multi-kinase inhibitor, the antimetabolite, or a combination thereof is administered locally to the affected eye in the form of a topical ophthalmic formulation.

13. The method according to claim 11, wherein the topical ophthalmic preparation is a solution, a suspension, or an emulsion.

14. The method according to claim 13, wherein the topical ophthalmic preparation further contains one or more pharmaceutically acceptable additives selected from stabilizers, surfactants, polymer-based carriers, gelling agents, organic auxiliary solvents, pH adjusting components, and osmotic pressure adjusting components, and may or may not contain a preservative.

15. The method according to claim 11, wherein the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant is inserted into the affected eye.

16. The method according to claim 15, wherein the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant further contains a pharmaceutically acceptable additive.

17. The sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant is Multi-kinase inhibitors, antimetabolites, or combinations thereof; and Biodegradable polymers selected from polylactic acid (PLA), polyglycolic acid (PLGA), and copolymers of polylactic acid and polyglycolic acid. The method according to claim 16, which contains

18. The method according to any one of claims 1 to 17, comprising administering the treatment to a patient with pterygium.

19. A method for alleviating congestion and symptoms in pterygium, pinguecula, and pseudopterygium in patients requiring treatment, comprising administering a therapeutically effective amount of a multikinase inhibitor to the affected eye.

20. A method for reducing or preventing pterygium recurrence in a subject requiring treatment, comprising administering a therapeutically effective amount of a multikinase inhibitor to the affected eye of the subject.

21. The method according to claim 20, wherein the administration is performed before surgical removal of the pterygium.

22. The method according to claim 20, wherein the administration is performed during a surgical removal procedure for pterygium.

23. The method according to claim 20, wherein the administration is performed after surgical removal of the pterygium.

24. The multi-kinase inhibitor is IC50 for VEGFR(1,2,3) is <50nM, The IC50 for PDGFR(α,β) is <100 nM, IC50 for FGFR(1,2,3) is <150nM IC50 for FGFR4 is <1000 nM, IC50 for FLT3 is <50nM, IC50 for Lck is <50 nM, IC50 for Lyn is <200 nm, The method according to any one of claims 19 to 23, wherein IC50 for Src is <200 nM.

25. The method according to claim 24, wherein the multikinase inhibitor is selected from the group consisting of nintedanib {(3Z)-3-{[(4-{methyl[(4-methylpiperazine-1-yl)acetyl]amino}phenyl)amino}amino}(phenyl)methylidene}-2-oxo-2,3-dihydro-1H-indole-6-carboxylate methyl}, its free base, hydrate, solvate, or pharmaceutically acceptable salt.

26. The method according to claim 25, wherein the multi-kinase inhibitor is nintedanib free base or nintedanib esylate (ethanesulfonate).

27. The method according to any one of claims 19 to 26, wherein the multikinase inhibitor is administered to the affected eye in the form of a topical ophthalmic preparation, a sustained-release semi-solid preparation, a sustained-release solid preparation, or an intraocular implant.

28. The method according to claim 27, wherein nintedanib is administered to the affected eye in the form of a topical ophthalmic formulation.

29. The method according to claim 28, wherein the topical ophthalmic preparation is a solution, a suspension, or an emulsion.

30. The method according to claim 28, wherein the concentration of nintedanib in the topical ophthalmic preparation is 0.001% to 10% by weight of the total amount of the preparation.

31. The method according to claim 27, wherein the topical ophthalmic preparation further contains one or more pharmaceutically acceptable additives selected from stabilizers, surfactants, polymer-based carriers, gelling agents, organic auxiliary solvents, pH adjusting components, osmotic pressure adjusting components, and preservatives.

32. The method according to claim 27, wherein the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant is inserted into the affected eye.

33. The method according to claim 32, wherein the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant contains nintedanib and a pharmaceutically acceptable additive.

34. The method according to claim 33, wherein the amount of nintedanib in the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant is 1 μg to 100 mg.

35. The method according to claim 27, wherein the sustained-release semi-solid preparation, sustained-release solid preparation, or intraocular implant contains nintedanib; and a biodegradable polymer selected from polylactic acid (PLA), polyglycolic acid (PLGA), and copolymers of polylactic acid and polyglycolic acid.

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