Treatment of dry eye syndrome using tanfanercept ophthalmic composition

Abstract

To provide an appropriate ophthalmic composition for treatment of dry eye syndrome using Tanfanercept.SOLUTION: Provided is an ophthalmic composition comprising Tanfanercept as an active ingredient for treating dry eye syndrome in a patient having moderate to severe dry eye.SELECTED DRAWING: Figure 26

Description

[Technical Field]

[0001] The present invention relates to the treatment of xerophthalmia with tamperenacept ophthalmic compositions. [Background technology]

[0002] Xerophthalmia is an eye condition that affects millions of people each year. It is particularly prevalent in postmenopausal women due to hormonal changes caused by sterilization. Xerophthalmia manifests in varying degrees from person to person. In mild cases, patients experience a burning, dry, and foreign body sensation, while in severe cases, vision can be severely impaired. Other diseases, such as Sjögren's syndrome and cicatricial pemphigoid, can also present with a dry eye complex.

[0003] Based on current research findings, it is understood that xerophthalmia is a disease that occurs when various stresses cause an autoimmune reaction involving cytokines and antigen-presenting cells on the ocular surface, leading to the concentration of immune cells in the corneal tissue, which then damages the tissue.

[0004] The typical treatment for dry eye syndrome involves the use of artificial tears to supplement the tear film, reducing tear evaporation and stabilizing the tear film. Alternatively, tear inserts may be used to stimulate internal tear production. The main components of artificial tears are cellulose ether, carbomer, polyvinyl alcohol, polyvinylpyrrolidone, or sodium hyaluronate, and are typically prepared in a buffer or isotonic saline solution. These components either increase the viscosity of the solution to prevent it from easily flowing from the eye, or act as a lubricant by preventing tear evaporation. However, these treatments are limited to treating the symptoms rather than providing a fundamental cure.

[0005] Meanwhile, it has become clear that the cause of dry eye is related to inflammatory reactions on the surface of the eye, and research is underway to apply various types of anti-inflammatory substances to treatment, and their effectiveness has been demonstrated.

[0006] Specifically, it has been reported that patients suffering from dry eye symptoms exhibit disproportionately excessive inflammatory properties in related ocular tissues, such as the lacrimal gland and meibomian gland, and various compounds, such as steroids, cytokine efflux inhibitors, cyclosporine A, and 15-HETE, are known to be effective in treating dry eye syndrome.

[0007] Tumor necrosis factor alpha (TNFα), a key factor involved in inflammatory responses, binds to human tumor necrosis factor receptors (TNF receptors, TNFR) I and II present on the cell surface, initiating various cellular responses, including cell death and inflammation. Since it was demonstrated that inhibiting the binding of TNFα to tumor necrosis factor receptors (TNFRs) can treat various autoimmune-related inflammatory diseases, various TNFα inhibitors have been developed. Representative TNFα inhibitors include etanercept (Enbrel), which combines soluble TNFRII with Fc, and the anti-TNFα antibodies infliximab (Remicade) and adalimumab (Humira). These are primarily used as treatments for rheumatoid arthritis, psoriasis, Crohn's disease, and other conditions.

[0008] However, this anti-TNF-α antibody formulation has a limitation in that its large molecular weight prevents the formulation from efficiently reaching inflammatory diseases induced at local sites. Therefore, the present applicant has developed a polypeptide molecule that is small in size and highly active, making it suitable for treating local inflammatory diseases. The TNF-α inhibitor of the present invention, a modified human tumor necrosis factor receptor-1 polypeptide, tampercept, is disclosed in the applicant's patent application, Korean Patent Publication No. 2012-0072323, and its use in treating dry eye syndrome is also described in Korean Patent Publication No. 2013-0143484. Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention aims to identify patient populations suitable for the treatment of xerophthalmia with tampernacept ophthalmic compositions.

[0010] The present invention provides suitable ophthalmic compositions for the treatment of xerophthalmia with tampercept, and methods for treating xerophthalmia therewith. [Means for solving the problem]

[0011] Tamperenacept is a polypeptide consisting of a total of 171 amino acids. As with general protein-containing pharmaceutical compositions, one of the most important challenges in drug development is to ensure stability and optimal efficacy before administration to patients. In addition to drug stability, another important issue in drug development is identifying the patient group most likely to respond to the drug and thereby enhance the drug's efficacy. Even a drug with proven efficacy does not necessarily work for all patients. Therefore, in order to provide patients with information that allows them to select the optimal drug for disease treatment, it is preferable to identify the appropriate patient group that can be treated with the drug.

[0012] After identifying the use of tampercept for dry eye syndrome, the inventors conducted extensive research to commercialize this drug as an ophthalmic composition for the treatment of dry eye syndrome. Given the characteristics of tampercept, a protein drug, the inventors studied the composition of an ophthalmic composition that could enhance the stability of the drug, and through clinical trials identified the patient group with the highest therapeutic efficacy when treated with tampercept.

[0013] As a result, the present invention provides an ophthalmic composition comprising tampanacept as an active ingredient for treating xerophthalmia in patients with moderate to severe dry eye.

[0014] The results of clinical trials of tamperacept confirmed that ophthalmic compositions containing tamperacept exhibited superior efficacy in treating dry eye in patients with moderate to severe dry eye than in patients with mild dry eye. This contradicts the fact that most ophthalmic compositions for treating dry eye have been evaluated as being more suitable for treating mild dry eye than moderate to severe dry eye. The fact that ophthalmic compositions containing tamperacept exhibit superior efficacy in treating dry eye in patients with moderate to severe dry eye than in patients with mild dry eye provides a substantially new therapeutic approach in the field of ophthalmic compositions for treating dry eye.

[0015] The patient with moderate to severe dry eye may be a patient with a corneal staining index of 2 or more in at least one area of ​​the inferior cornea, central cornea, and superior cornea, and a Schirmer test score of 1 or more and 7 or less.

[0016] In the present invention, signs, which are objective evaluation indicators for xerophthalmia, and symptoms, which are subjective evaluation indicators, are used to evaluate the degree of dry eye in patients with dry eye and the degree of improvement of dry eye caused by medication.

[0017] The Corneal Staining Score (CSS) was used as an objective evaluation index. The CSS method involves assessing the degree of corneal damage after applying a stain to the eye and assigning a score. The CSS is expressed as a score between 0 and 4, with the greater the degree of corneal damage, the greater the staining. A CSS score of 2 indicates mild, 3 moderate, and 4 severe. Even if a CSS score is 2 for any part of the eye, other objective signs and subjective evaluation indexes may indicate moderate or severe dry eye. Therefore, a combined assessment is made based on other evaluation indices.

[0018] The objective evaluation index measured along with the corneal staining index is the Schirmer test score. The Schirmer test (STT) is a method in which a Schirmer test strip is placed on the margin of the lower eyelid of each patient's eye, and after 5 minutes of closing the eyes, the Schirmer strip is removed and the length (mm) of the moist area is measured and recorded. In the present invention, a patient with moderate to severe dry eye syndrome may have a corneal staining index of 2 or more in at least one of the inferior, central, and superior corneal regions and a Schirmer test score of 1 to 7, but a lower Schirmer test score indicates a more severe dry eye syndrome.

[0019] When an ophthalmic composition according to the present invention is administered to a patient suffering from dry eye, improvement in moderate to severe dry eye symptoms can begin to appear within 8 weeks of administration of the ophthalmic composition.

[0020] The period of 8 weeks after administration is an exemplary period, and improvement in symptoms may occur within a shorter period of time, such as 7, 6, 5, 4, 3, 2, or 1 week after administration. This may vary from individual, i.e., patient to patient, and can vary depending on various factors such as the severity or the dose of the drug, but it means that improvement in symptoms will occur within at least 8 weeks after administration.

[0021] In particular, the improvement of symptoms may be measured by a decrease in the overall corneal staining index and the central corneal staining index. Although the areas with a high corneal staining index may vary depending on the patient or the severity of dry eye, the overall corneal staining index includes all the corneal staining indices of each area, and therefore a decrease in the overall corneal staining index has a significant meaning in improving symptoms. Generally, when an ophthalmic composition is administered in the form of eye drops, it remains in the lower part of the cornea, and is more likely to improve the lower corneal staining index. However, in the present invention, a decrease in the central corneal staining index is measured as an indicator of improvement of symptoms.

[0022] On the other hand, whether or not moderate to severe dry eye symptoms have improved can also be evaluated based on the improvement of symptoms, which is a subjective evaluation index.

[0023] When the ophthalmic composition according to the present invention is administered to patients with dry eye syndrome, improvement in moderate to severe dry eye symptoms can begin to appear within 8 weeks of administration of the ophthalmic composition.

[0024] The period of 8 weeks after administration is an exemplary period, and symptomatic improvement may occur within a shorter period, such as 7, 6, 5, 4, 3, 2, or 1 week after administration. This may differ for each individual, i.e., patient, and may vary depending on various factors such as the severity or the dosage of the drug, but it means that symptomatic improvement will occur within at least 8 weeks after administration.

[0025] The improvement of the symptoms may be measured by a decrease in the eye dryness score (EDS). The eye dryness score used in the present invention is an index of the eye dryness item in the visual analog scale (VAS). Subjects evaluate the eye dryness index using a number from 0 to 100, with 0 corresponding to "no dryness" and 100 corresponding to "extreme dryness."

[0026] In a specific embodiment of the present invention, the patient may have a central corneal staining index of 2 or greater (CCSS≧2) before administration of the ophthalmic composition. CCSS≧2 means that the central corneal staining index is at or above the mild level. Alternatively, the patient may have a total corneal staining index of 5 or greater (TCSS≧5) before administration of the ophthalmic composition. TCSS≧5 means that the sum of the lower, central, and upper corneal staining indexes is at or above the mild level.

[0027] In another embodiment of the present invention, the patient may have a Schirmer test score of 1 or more and 7 or less before administration of the ophthalmic composition. In yet another embodiment, the patient may have a Schirmer test score of 1 or more and 3 or less before administration of the ophthalmic composition. As described above, a lower Schirmer test score indicates a more severe dry eye condition. The ophthalmic composition according to the present invention exhibits excellent therapeutic effects for dry eye even in patients with a Schirmer test score of 1 or more and 3 or less.

[0028] In yet another specific example, the patient may have an ocular discomfort index (ODS) of 3 or greater before administration of the ophthalmic composition. The ocular discomfort index is a score of 0 to 4 that indicates the degree of ocular discomfort the patient experiences subjectively, with higher scores indicating greater ocular discomfort. An ocular discomfort index (ODS) of 3 or greater means that the patient experiences intermittent or persistent discomfort in the eye, with moderate to severe dryness.

[0029] In yet another embodiment of a patient population suitable for administration of the ophthalmic composition of the present invention, the patient may have a Total Corneal Staining Score (TCSS) of 5 or greater and an Ocular Discomfort Index (ODS) of 3 or greater prior to administration of the ophthalmic composition.

[0030] In addition, patients suitable for administration of the ophthalmic composition of the present invention may be patients with a total corneal staining index (TCSS) of 4 or more and an eye dryness index (EDS) of 40 or more before administration of the ophthalmic composition, for example, patients with a total corneal staining index (TCSS) of 5 or more and an eye dryness index (EDS) of 60 or more.

[0031] Furthermore, patients suitable for administration of the ophthalmic composition of the present invention may be patients who have used artificial tears within one month prior to administration of the ophthalmic composition.

[0032] The ophthalmic composition according to the present invention may be formulated in the form of eye drops.

[0033] Specifically, the ophthalmic composition according to the present invention may be administered in the form of eye drops once or more times a day, for example, the ophthalmic composition may be administered in the form of eye drops twice a day.

[0034] Meanwhile, the present inventors have conducted extensive research into the stability of tampercept in order to develop an ophthalmic composition that can minimize the formation of tampercept-derived impurities (acidic / basic variants) not only under refrigerated storage conditions but also during storage under accelerated and harsh conditions. In particular, the present inventors have conducted research into various formulations, including buffers, isotonicity agents, pH ranges, and functional excipients. As a result, the present inventors have found that the use of stabilizers such as histidine and sucrose actually produces acidic / basic variants at certain pH levels, thereby affecting the stability of tampercept compositions. Therefore, the present invention provides a stabilizer-free tampernacept ophthalmic composition that is stable at pH 5.0 to pH 6.5.

[0035] Specifically, the present invention provides a stable tampercept-containing ophthalmic composition that contains tampercept and a buffer system of pH 5.0 to pH 6.5 and is substantially free of stabilizers.

[0036] Tamperecept is a TNFRI variant disclosed in Korean Patent Publication No. 2013-0143484, and is represented by an amino acid sequence containing amino acid mutations L68V / S92M / H95F / R97P / H98G / K161N in the amino acid sequence (TNFRI171) consisting of amino acid positions 41 to 211 of the native TNFRI sequence.

[0037] Since tampercept is a polypeptide consisting of a total of 171 amino acids, one of the most important challenges in drug development is to ensure its stability and optimal efficacy before administration to patients, as is the case with general protein-containing pharmaceutical compositions.

[0038] To develop a formulation that can ensure the stability of tamperacept, the inventors first tested the storage stability of tamperacept and observed that tamperacept formed charge variants during storage (Experimental Example 1). As used herein, the term "charge variant" refers to a protein or polypeptide that has been modified from its native state, resulting in a different charge. In one example, a charge variant is more acidic than the native protein or polypeptide, i.e., has a lower pI value than the native protein or polypeptide. In another example, a charge variant is more basic than the native protein or polypeptide, i.e., has a higher pI value than the native polypeptide. Such modifications may be engineered or the result of natural processes, such as oxidation, deamidation, C-terminal processing of lysine residues, N-terminal pyroglutamate formation, and non-enzymatic glycation. In one example, a protein or polypeptide charge variant is a glycoprotein in which the glycans attached to the protein have been modified, for example, by the addition of sialic acid or its derivatives, resulting in a different charge of the glycoprotein compared to the parent glycoprotein. As used herein, a "tampercept charge variant" is a substance that is modified from the native state of tampercept and that differs from the charge of tampercept.

[0039] Charge variants are generally known to reduce the activity of drugs, so it is necessary to control the amount of charge variants produced below a certain level. Therefore, the inventors investigated whether it is possible to minimize the production of impurities such as charge variants using ingredients that can be used as ophthalmic stabilizers, and initially determined that sucrose and histidine are preferred (Experimental Example 2). However, in further research into the optimal pH for tampanacept-containing compositions, they found that the rate of charge variant production was high even with the use of these stabilizers at certain pH levels. Rather, they found that adjusting the pH to a level between pH 5.0 and pH 6.5 without using stabilizers was the best way to minimize the rate of charge variant production (Experimental Example 3).

[0040] Therefore, the present invention provides a tampercept-containing ophthalmic composition that contains tampercept and a buffer system of pH 5.0 to pH 6.5 and is substantially free of stabilizers.

[0041] It is preferable to include tamperenacept in the composition at an appropriate content, because a higher content may increase the content of impurities such as aggregates. In the tamperenacept-containing ophthalmic composition of the present invention, tamperenacept may be included in a content of 0.01% (w / v) to 1% (w / v), for example, 0.02% (w / v) to 1% (w / v), 0.05% (w / v) to 0.8% (w / v), 0.1% (w / v) to 0.7% (w / v), or 0.2% (w / v) to 0.6% (w / v). Considering commercial purposes, tamperenacept may be included in the composition at a content of 0.25% (w / v), 0.5% (w / v), 1% (w / v), or the like. The content of tamperenacept may vary depending on the type and severity of the disease of the patient to be administered.

[0042] The tampercept-containing ophthalmic composition according to the present invention comprises a buffer system having a pH of 5.0 to 6.5. The buffer system may have a pH of 5.0 to 6.5, and all numerical ranges within the range of pH 5.0 to 6.5, such as a buffer system of pH 5.0 to 6.0, a buffer system of pH 5.5 to 6.5, a buffer system of pH 5.5 to 6.0, and a buffer system of pH 5.8 to 6.3, are included in the category of the present invention. In one specific example of the present invention, the tampercept-containing ophthalmic composition according to the present invention comprises a buffer system of pH 5.0 to 6.0. In another specific example of the present invention, the tampercept-containing ophthalmic composition according to the present invention comprises a buffer system of pH 5.5 to 6.0.

[0043] Methods for implementing a buffer system in a tampercept-containing ophthalmic composition according to the present invention are well known to those skilled in the art. A buffer system having a pH of 5.0 to 6.5 may contain one or more buffers selected from the group consisting of phosphate buffer, histidine buffer, acetate buffer, succinate buffer, citrate buffer, glutamate buffer, and lactate buffer. It is believed that the stability of tampercept can be ensured regardless of the buffer system used, as long as the pH range of 5.0 to 6.5 is satisfied. However, the use of a specific buffer system may be relatively preferable. In the following examples, it was confirmed that citrate buffer is relatively advantageous over acetate buffer in terms of controlling the formation of aggregates and charge variants (Experimental Example 4). Therefore, in certain embodiments of the present invention, the buffer system may be a buffer system containing a citrate buffer, for example, a citrate buffer system or a citrate-phosphate buffer system, but is not limited thereto. The buffering agent included in the buffer system may be composed of a conjugate acid-conjugate base combination to enhance the buffering effect. For example, in one embodiment of the present invention, the buffer system includes a sulphate buffer, wherein the sulphate buffer includes trisodium sulphate (conjugate base) and citric acid (conjugate acid).

[0044] The buffer system of the present invention contains a buffering agent at a concentration of 5 mM to 50 mM, for example, a buffering agent at a concentration of 10 mM to 30 mM.

[0045] The ophthalmic composition of the present invention is characterized by being substantially free of stabilizers. Here, stabilizers refer to additional components contained in the formulation to prevent a decrease in the chemical, physical stability, or biological activity of tamperenacept, which is used as an active ingredient. For example, it is well known that ophthalmic compositions use sugars such as sucrose and mannitol to inhibit protein aggregation, or amino acid stabilizers such as proline, arginine, glycine, lysine, or methionine to stabilize proteins. As confirmed in the following examples, unlike typical cases, the inclusion of such stabilizers has been found to adversely affect the stability of tamperenacept, and therefore the present invention is substantially free of stabilizers. The buffers and tonicity agents described in the present invention are not included in the stabilizers.

[0046] By "substantially free" of stabilizers is meant less than 0.1% (w / v), less than 0.05% (w / v), less than 0.03% (w / v), less than 0.02% (w / v), less than 0.01% (w / v), less than 0.005% (w / v), less than 0.001% (w / v), or most preferably, no stabilizers at all.

[0047] The osmolality of the ophthalmic composition may be 260 mOsm / kg to 320 mOsm / kg.

[0048] The ophthalmic composition of the present invention may further contain an isotonicity agent in addition to the active ingredient tampercept and a buffer system. The isotonicity agent is used to adjust the osmotic pressure of the ophthalmic composition of the present invention. In the present invention, the isotonicity agent is included so that the ophthalmic composition of the present invention has an osmotic concentration of 260 mOsm / kg to 320 mOsm / kg. Osmotic pressure is measured by the number of particles dissolved per unit of water. The fewer the number of solute particles in a solution relative to the number of water units (solvent), the less concentrated the hypo-osmotic solution. When solutions with different solute concentrations are separated using a semi-permeable membrane (a membrane that allows only solvent molecules to pass), osmosis occurs, where solvent molecules cross the membrane from a low concentration to a high concentration, forming a concentration equilibrium. The pressure that drives this movement is called osmotic pressure and depends on the number of solute "particles" in the solution. Solutions containing the same particle concentration and subjected to the same osmotic pressure are called iso-osmotic. If a hypo-osmotic or hyper-osmotic solution is placed in the eye, it may damage the eye, therefore, an iso-osmotic solution is necessary for drugs used in the eye. In the present invention, sodium chloride is used as the tonicity agent. The content of the tonicity agent in the ophthalmic composition according to the present invention may be 0.5% (w / v) to 1% (w / v). The concentration of the tonicity agent in the ophthalmic composition according to the present invention may be 100 mM to 150 mM.

[0049] In one embodiment of the present invention, the ophthalmic composition according to the present invention comprises tamperacept, a buffer system having a pH of 5.5 to 6.0, a tonicity agent, and water. In one example, the ophthalmic composition according to the present invention comprises tamperacept, a buffer system having a pH of 5.5 to 6.0 containing a citrate buffer, sodium chloride, and water.

[0050] The tamperenacept-containing ophthalmic compositions according to the present invention are highly stable even under accelerated or harsh conditions.

[0051] The tamperenacept-containing ophthalmic composition according to the present invention may have an amount of charge variants of 20% or less after 6 months of storage under accelerated conditions.

[0052] The tamperenacept-containing ophthalmic composition according to the present invention may have an amount of basic variant of 10% or less after 6 months of storage under accelerated conditions.

[0053] The tamperenacept-containing ophthalmic compositions of the present invention have an amount of acidic variant of 10% or less after 6 months of storage under accelerated conditions.

[0054] The tampercept-containing ophthalmic composition according to the present invention has an amount of charge variants of tampercept of 20% or less after 36 months of storage under long-term storage conditions.

[0055] The tampercept-containing ophthalmic composition according to the present invention has an amount of basic variant of tampercept of 10% or less after 36 months of storage under long-term storage conditions.

[0056] The tampercept-containing ophthalmic composition according to the present invention has an amount of acidic variant of tampercept of 10% or less after 36 months of storage under long-term storage conditions.

[0057] The present invention further provides a method for treating xerophthalmia, which comprises administering to a patient with moderate to severe dry eye an ophthalmic composition containing tampanacept as an active ingredient.

[0058] The ophthalmic composition containing tamperacept as an active ingredient is as described in the specific example above, and the patients suitable for administration of the ophthalmic composition are also as described above.

[0059] The ophthalmic pharmaceutical composition according to the present invention may be administered by a conventional method, such as intravenous administration, to patients suffering from TNF-mediated eye diseases such as xerophthalmia, by improving the physicochemical and biological stability of tamperacept. [Effects of the Invention]

[0060] According to the present invention, a tamperacept-containing ophthalmic composition has demonstrated excellent efficacy in improving the clinically important central and global corneal staining index in clinical trials for dry eye syndrome. Furthermore, the improvement in the central and global corneal staining index was greater in patients with moderate to severe dry eye, and the degree of improvement was also greater in patients with higher subjective symptoms such as ocular discomfort index and dry eye index. Despite using the same dose of tamperacept, the tamperacept-containing ophthalmic composition demonstrated superior efficacy in patients with more severe signs and symptoms, an unexpected finding. This provides a novel therapeutic alternative in the current situation where there is no treatment for moderate to severe dry eye syndrome. [Brief explanation of the drawings]

[0061] [Figure 1] 1 shows the results of isoelectric focusing (IEF) of tampernacept after storage at 37°C for 0 to 4 weeks. [Figure 2] 1 shows the results of IEX-HPLC analysis of tamperacept after storage at 37°C for 0 to 4 weeks. [Figure 3] IEF analysis of charge mutants is shown. [Figure 4] The results of IEX-HPLC analysis of charge mutants are shown. [Figure 5] The figure shows the change in the IEX-HPLC main peak for a control group containing no stabilizer and stabilizer screening solution groups containing methionine, glycine, histidine hydrochloride, and sucrose as stabilizers. [Figure 6] The figure shows the change in IEX-HPLC acidic mutants in a control group containing no stabilizer and stabilizer screening solution groups containing methionine, glycine, histidine hydrochloride, and sucrose as stabilizers. [Figure 7] The graph shows the change in IEX-HPLC basic mutants in a control group containing no stabilizer and stabilizer screening solution groups containing methionine, glycine, histidine hydrochloride, and sucrose as stabilizers. [Figure 8]1 shows the results of analyzing the stability of tamperacept ophthalmic compositions under various pH and stabilizer conditions after storage at 40° C. for 4 weeks using RP-HPLC. [Figure 9] 1 shows the stability of tamperacept ophthalmic compositions after storage at 40° C. for 4 weeks under various pH and stabilizer conditions, as shown by the change in the main peak of IEX-HPLC. [Figure 10] 1 shows the stability of tamperacept ophthalmic compositions after storage at 40° C. for 4 weeks under various pH and stabilizer conditions, as shown by the change in the amount of acidic variants measured by IEX-HPLC. [Figure 11] 1 shows the stability of tamperacept ophthalmic compositions after storage at 40° C. for 4 weeks under various pH and stabilizer conditions, as shown by the change in IEX-HPLC basic variants. [Figure 12] The results of SEC-HPLC analysis of four types of tamperacept ophthalmic compositions stored at 4°C, 25°C, and 40°C are shown. [Figure 13] The results of SEC-HPLC analysis of four types of tamperacept ophthalmic compositions stored at 4°C, 25°C, and 40°C are shown. [Figure 14] The results of SEC-HPLC analysis of four types of tamperacept ophthalmic compositions stored at 4°C, 25°C, and 40°C are shown. [Figure 15] 1 shows the results of an analysis of acidic variants using IEX-HPLC performed on four types of tamperacept ophthalmic compositions while storing them under conditions of 4°C, 25°C, and 40°C. [Figure 16] 1 shows the results of an analysis of acidic variants using IEX-HPLC performed on four types of tamperacept ophthalmic compositions while storing them under conditions of 4°C, 25°C, and 40°C. [Figure 17] 1 shows the results of an analysis of acidic variants using IEX-HPLC performed on four types of tamperacept ophthalmic compositions while storing them under conditions of 4°C, 25°C, and 40°C. [Figure 18] 1 shows the results of basic charge variant analysis using IEX-HPLC performed on four types of tamperacept ophthalmic compositions while storing them under conditions of 4° C., 25° C., and 40° C. [Figure 19] 1 shows the results of basic charge variant analysis using IEX-HPLC performed on four types of tamperacept ophthalmic compositions while storing them under conditions of 4° C., 25° C., and 40° C. [Figure 20] 1 shows the results of basic charge variant analysis using IEX-HPLC performed on four types of tamperacept ophthalmic compositions while storing them under conditions of 4° C., 25° C., and 40° C. [Figure 21] The stability of tamperacept ophthalmic compositions is demonstrated through aggregate analysis using SEC-HPLC. [Figure 22] The stability of tamperacept ophthalmic compositions is demonstrated through charge variant analysis using IEX-HPLC. [Figure 23] The stability of tamperacept ophthalmic compositions is demonstrated through charge variant analysis using IEX-HPLC. [Figure 24] 1 is a schematic diagram of a clinical trial schedule for the tamperacept ophthalmic composition of the present invention, targeting patients with dry eye syndrome. [Figure 25] This is a diagram showing the method for evaluating the corneal staining index. [Figure 26] This shows the change in the inferior, superior, and central corneal values, as well as the total of these, for the entire cornea (total) from baseline to the end of medication (week 8) for randomly assigned subjects (Intent-to-Treat Population, ITT Population). [Figure 27] The mean change in CCSS scores over 8 weeks for all subjects is shown. [Figure 28] The mean change in TCSS index over 8 weeks for all subjects is shown. [Figure 29] Shown is the mean change in EDS from baseline at Week 8 for subjects who had used artificial tears within one month of Visit 1. [Figure 30] The mean change in CCSS scores according to disease severity at the 8-week baseline is shown. [Figure 31] Mean change in TCSS according to disease severity from baseline to 8 weeks is shown. DETAILED DESCRIPTION OF THE INVENTION

[0062] The advantages, features, and methods for achieving the same of the present invention will become apparent from the following examples, examples, and experimental examples, which are provided to aid in the understanding of the present invention and are not intended to limit the scope of the present invention.

[0063] [Part A] Tamperacept Ophthalmic Composition Experimental Example 1: Analysis of Tamperacept Charge Variants Because the generation of charge variants can affect drug activity, stability, and safety, we first analyzed the charge variants of tamperacept. After storing tamperacept at 37°C for 4 weeks, we performed isoelectric focusing (IEF) and IEX-HPLC analysis.

[0064] Isoelectric focusing 10 μg was loaded per well on a gel ranging from pH 3.0 to pH 7.0, and electrophoresis was performed at 100 V for 1 hour, 200 V for 1 hour, and 500 V for 30 minutes. After fixation with 12% trichloroacetic acid for 30 minutes, the sample was stained with Coomassie blue.

[0065] IEX-HPLC (ion exchange high performance liquid chromatography) Ion-exchange high-performance liquid chromatography (IEX-HPLC) uses an ion exchanger to separate proteins based on their affinity for the stationary phase of the column, which is determined by their net charge. This test method was performed using a cation-exchange column and a high-performance liquid chromatograph (HPLC) equipped with a temperature controller (set at 25°C), an automatic sample extractor (set at 4°C), a UV detector driven at 280 nm, and a flow rate of 0.7 mL / min.

[0066] Separation and purification of charge variants To separate and analyze the charge variants present in the tamperacept ophthalmic composition based on their characteristics, separation and purification was performed according to salt concentration using an SP-HP column and fast particle chromatography (FPLC). Based on the characteristics of the charge variants, samples were prepared as acidic variants (A), main peak samples (B), and basic variants (C), and each was analyzed by IEF and IEX-HPLC.

[0067] FIG. 1 shows the results of isoelectric focusing (IEF) of tampernacept after storage at 37° C. for 0 to 4 weeks.

[0068] As a result of isoelectric focusing, as shown in Figure 1A, the bands at lower pI values ​​became stronger with increasing storage time, clearly confirming the generation of acidic variants of tampercept.

[0069] FIG. 2 shows the results of IEX-HPLC analysis of tamperacept after storage at 37° C. for 0 to 4 weeks.

[0070] The results of IEX-HPLC also confirmed that the amount of acidic variants increased with increasing storage time, as shown in Figure 2 .

[0071] To investigate the characteristics of the charge variants, the acidic variant, the main peak sample, and the basic variant sample obtained by charge separation using an SP-HP column were analyzed by IEF and IEX-HPLC.

[0072] Figure 3 shows the results of IEF analysis of the charge variants, and Figure 4 shows the results of IEX-HPLC analysis of the charge variants. As can be seen from Figures 3 and 4, Sample A, which separated only the acidic variants, eluted earlier than the main peak sample in the chromatogram, and the IEF results also showed that it had a lower pI than the main peak sample. In addition, Sample B, which is a basic variant sample, eluted later than the main peak sample in the chromatogram, and the IEF results also showed that it had a somewhat higher pI value, including the main peak sample.

[0073] Experimental Example 2: Screening of ophthalmic stabilizers

[0074] Stabilizers added to protein compositions are necessary to stably preserve the formulation before administration to patients. These stabilizers minimize impurities, such as aggregates and charge variants, that may be induced during storage, thereby maintaining a stable formulation during storage. Therefore, it is of utmost importance to select an appropriate stabilizer that stabilizes the main component of a protein composition to produce a stable composition. The present inventors first conducted the following experiment to conduct a stress test (storage at 40°C for 4 weeks) on each stabilizer to select the type of stabilizer that stabilizes the main component, tampanacept.

[0075] 1) Preparation of tampercept solution sample A 10 mg / mL solution of tamperacept in 20 mM sodium citrate buffer containing 125 mM sodium chloride was prepared.

[0076] 2) Preparation of buffer pH 7.0, 20 mM citrate phosphate 0.37 g of citric acid anhydrous and 2.58 g of disodium hydrogen phosphate were added to 900 mL of ultrapure water and mixed thoroughly. The solution was titrated to pH 7.0 with 37% hydrochloric acid or 40% sodium hydroxide, and then ultrapure water was added to make a final volume of 1 L.

[0077] 3) Preparation of stabilizer screening solution Four stabilizers (methionine 0.149 g, glycine 0.751 g, histidine hydrochloride 1.55 g, sucrose 6.84 g) were added to 100 mL of the buffer prepared in 2) to prepare four stabilizer screening compositions at pH 7.0.

[0078] [Table 1]

[0079] 4) Sample preparation and evaluation Tampercept and 10 mL of the ophthalmic stabilizer screening solution prepared in 3) were added to a 3.5 kDa centrifugal filter, and the sample was centrifuged at 4°C and 4000 rpm. The tampercept buffer in 1) was replaced with the stabilizer screening buffer in 3). The above process was repeated to prepare a stabilizer screening solution (sample) containing 1 mg / mL of tampercept. This was stored at 40°C for 4 weeks, and the samples at 0 and 4 weeks were analyzed by IEX-HPLC to analyze the physicochemical impurities detected in each sample.

[0080] [Table 2]

[0081] Table 2 and Figures 5 to 7 show the results of IEX-HPLC analysis of a control group containing no stabilizer and stabilizer screening solution groups containing methionine, glycine, histidine hydrochloride, and sucrose as stabilizers, respectively.

[0082] As a result, as shown in Table 2 and Figures 5 to 7, it was confirmed that when sucrose and histidine hydrochloride were added, the amount of basic and acidic mutants produced tended to decrease. In particular, it was confirmed that histidine hydrochloride significantly reduced the amount of acidic mutants produced compared to the control group and other stabilizers.

[0083] Experimental Example 3: Preparation and Stability Evaluation of Ophthalmic Compositions with Various pHs Since the pH of tears is 7.0 to 7.5, it is most preferable to prepare an ophthalmic composition under similar pH conditions. However, since protein stability can be significantly affected by pH, ophthalmic compositions with various pH values ​​were prepared and their stability was evaluated.

[0084] (1) Manufacture of ophthalmic compositions 1) Preparation of buffer pH 5.0 to pH 7.0, 20 mM sodium citrate Citric acid anhydrous and disodium hydrogen phosphate were added to 400 mL of ultrapure water and mixed thoroughly to adjust the pH. (For pH 5.0 / 5.5 buffer preparation: 0.62 g of citric acid anhydrous, 0.97 g of disodium hydrogen phosphate. For pH 6.0 / 6.5 buffer preparation: 0.43 g of citric acid anhydrous, 1.11 g of disodium hydrogen phosphate. For pH 7.0 buffer preparation: 0.19 g of citric acid anhydrous, 1.29 g of disodium hydrogen phosphate.) The solution was titrated to pH 5.0-7.0 using 37% hydrochloric acid or 40% sodium hydroxide, and then ultrapure water was added to make the final volume of 500 mL.

[0085] 2) Preparation of ophthalmic compositions containing stabilizers (pH 5.0 to pH 7.0) Five 20 mM sodium citrate buffers containing 200 mM sucrose and 1.55 g of histidine were added to 100 mL of each of the five buffers corresponding to pH 5.0 to pH 7.0 prepared in 1), yielding five 20 mM sodium citrate buffers containing 200 mM sucrose and five 20 mM sodium citrate buffers containing 100 mM histidine. The 20 mM sodium citrate buffers containing pH 5.0 to pH 7.0 prepared in 1) were used as is for the stabilizer-free experimental group, yielding a total of 15 buffers containing and not containing stabilizers, yielding pH 5.0 to pH 7.0.

[0086] 3) Sample preparation and evaluation Using a 3.5 kDa centrifugal filter, 4 mL of the buffer prepared in 2) and tampercept were added, and the sample was centrifuged at 4°C and 4000 rpm to replace the existing tampercept buffer with a buffer containing a stabilizer ranging from pH 5.0 to pH 7.0. This process was repeated to prepare 15 samples with different pHs and stabilizers. These were stored at 40°C for 4 weeks, and samples were analyzed at 0 and 4 weeks. Physicochemical impurities detected in each sample were analyzed.

[0087] [Table 3]

[0088] (2) Evaluation of tamperacept stability by pH and stabilizers 1) Analysis by reversed-phase chromatography RP-HPLC (reverse-phase chromatography) is a method for assessing protein purity based on its polarity. This test method was performed using a high-performance liquid chromatograph (HPLC) equipped with a reverse-phase chromatography column, a temperature controller (set at 60°C), an automatic sample extractor (set at 4°C), a UV detector driven at 214 nm, and a flow rate of 1.0 mL / min.

[0089] The 15 samples prepared above were stored under harsh conditions (storage at 40°C) for 4 weeks, and the samples at week 0 and week 4 were analyzed to compare the change in mutant formation observed in each sample. As a result, as shown in Table 4 and Figure 8, at pH 7.0, the experimental groups containing sucrose or histidine as a stabilizer showed a lower mutant formation rate than the experimental group containing no stabilizer. However, the experimental group containing histidine as a stabilizer showed a higher mutant formation rate than the control group at all pH levels from 5.0 to 6.5, and the experimental group containing sucrose as a stabilizer showed a higher mutant formation rate than the control group at pH levels from 5.0 to 6.0.

[0090] [Table 4]

[0091] 2) Analysis by ion-exchange high-performance liquid chromatography Ion-exchange high-performance liquid chromatography (IEX-HPLC) uses an ion exchanger to separate proteins based on their affinity for the stationary phase of the column, which is determined by their net charge. This test method was performed using a cation-exchange column and a high-performance liquid chromatograph (HPLC) equipped with a temperature controller (set at 25°C), an automatic sample extractor (set at 4°C), a UV detector driven at 280 nm, and a flow rate of 0.7 mL / min.

[0092] The 15 samples previously prepared were stored under harsh conditions (40°C) for 4 weeks, and the samples at week 0 and week 4 were analyzed to compare the changes in the production of acidic / basic mutants observed in each sample.

[0093] As shown in Table 5 and Figures 9 to 11, the stabilizer-free group showed better results than the sucrose- or histidine-containing experimental groups at pH 5.0 to pH 6.5 in terms of the amount of acidic mutants produced. In particular, the stabilizer-free experimental group showed a change in the amount of acidic mutants produced of less than 10% at pH 5.0 to pH 6.0. For basic mutants, the stabilizer-free group showed better results than the sucrose- or histidine-containing experimental groups at pH 5.0 to pH 6.0. At pH 6.5 to pH 7.0, the sucrose-containing experimental group showed the lowest change in the amount of basic mutants produced, and the histidine-containing experimental group showed no significant difference from the stabilizer-free group.

[0094] [Table 5]

[0095] Analysis of the results showed that at pH 7.0, the sucrose- and histidine-containing groups tended to produce lower amounts of mutants than the stabilizer-free group, but when the pH was lowered to pH 5.5 or pH 6.0, the sucrose- and histidine-containing groups tended to produce significantly higher amounts of mutants than the stabilizer-free group.

[0096] A comprehensive analysis of the experimental results revealed that, contrary to expectations, the tamperacept ophthalmic composition without stabilizer showed the lowest amount of mutants produced at pH 5.5 to pH 6.0.

[0097] Generally, ophthalmic compositions are considered to be set at a pH of 7.0 to resemble the pH conditions in the body. However, since the stability of the active ingredient in an ophthalmic composition is an important factor in demonstrating its efficacy, it has been concluded that the tampernacept ophthalmic composition of the present invention preferably has a pH of 5.5 to 6.0 and does not contain sucrose or histidine.

[0098] Experimental Example 4: Evaluation of stability with buffer systems The tampercept concentration, pH, sodium chloride concentration, osmolality, etc. were fixed, and four groups were formed to compare the differences between the buffer composition and the presence or absence of two functional excipients, and screening to select the final formulation was carried out at 4°C, 25°C, and 40°C. The pH of the ophthalmic composition was fixed at 5.5 in consideration of the results of previous experiments, and the stability of tampercept in the buffer system was evaluated using sodium acetate (20 mM) and sodium citrate (20 mM) as basic buffers that can generally be used within this range.

[0099] Four tampercept ophthalmic compositions were prepared and tested for tampercept stability, as shown in Table 6 below.

[0100] [Table 6]

[0101] A stability test of the tamperacept ophthalmic composition was conducted using SEC-HPLC for aggregate analysis and IEX-HPLC for charge variant analysis. The stability test was conducted for a total of two months at 4°C, 25°C, and 40°C, and aggregate analysis using SEC-HPLC and charge variant analysis using IEX-HPLC were performed at 0 weeks, 2 weeks, 1 month, and 2 months.

[0102] (1) Aggregate Analysis Using SEC-HPLC SEC-HPLC (size-exclusion high-performance liquid chromatography) separates proteins by size by injecting a sample into a column filled with a porous gel-based stationary phase. This test method was performed using a size-exclusion column, a high-performance liquid chromatograph (HPLC) with a temperature controller (set at 25°C), an automatic sample extractor (set at 4°C), a UV detector driven at 214 nm, and a flow rate of 0.5 mL / min.

[0103] Tables 7-9 and Figures 12-14 show the results of SEC-HPLC analysis of the four tamperacept ophthalmic compositions stored at 4°C, 25°C, and 40°C. The SEC-HPLC analysis showed a tendency for aggregates to increase with increasing temperature, with rapid formation occurring particularly in formulations using acetate buffer. The three formulations using acetate buffer showed high FFS3 values ​​at 4°C and 2 months. The formulation without FFS2 excipients showed the most stable results at 25°C and 40°C for 2 weeks, 1 month, and 2 months.

[0104] [Table 7]

[0105] [Table 8]

[0106] [Table 9]

[0107] (2) Analysis of Acidic Mutants Using IEX-HPLC Tables 10 to 12 and Figures 15 to 17 show the results of the analysis of acidic variants by IEX-HPLC performed on the four types of tamperenacept ophthalmic compositions while storing them at 4°C, 25°C, and 40°C.

[0108] [Table 10]

[0109] [Table 11]

[0110] [Table 12]

[0111] (3) Analysis of basic mutants using IEX-HPLC Tables 13 to 15 and Figures 18 to 20 show the results of basic charge variant analysis using IEX-HPLC, which was carried out while storing the four types of tamperenacept ophthalmic compositions at 4°C, 25°C, and 40°C.

[0112] [Table 13]

[0113] [Table 14]

[0114] [Table 15]

[0115] IEX-HPLC analysis showed that all formulations showed a tendency for an increase in acidic and basic charge variants under high-temperature storage conditions at 40°C, and the acetate buffer (FFS1) formulation showed a pattern in which basic variants clearly increased at all temperatures.

[0116] In conclusion, we confirmed that acetate buffer (FFS1) exhibited relatively lower stability compared to citrate buffers (FFS2-4) in terms of aggregate and basic variant formation. Furthermore, when comparing the citrate buffer composition with and without functional excipients, the group without excipients showed the highest stability. Taking into consideration the simplification of the product manufacturing process, contamination during the manufacturing process, and issues of uniformity, we selected a dosage form using citrate buffer (FFS2) as the final dosage form. However, considering that the physiological pH of tears is neutral, we selected pH 6.0 as the pH of the final product, as this is expected to provide acceptable material stability and higher patient compliance.

[0117] Preparation Example 1: Preparation of tamperacept ophthalmic composition A buffer was prepared by dissolving 5.35 g of trisodium citrate dihydrate, 0.35 g of anhydrous citric acid, and 7.3 g of sodium chloride in 900 ml of ultrapure water. The pH of the prepared buffer was confirmed to be 6.0 ± 0.1, and then the solution was adjusted to a final volume of 1 L using a measuring cylinder and filtered through a 0.22 μm bottle top filter system.

[0118] Tamperacept was added to the prepared buffer to prepare a 0.25% tamperacept eye drop composition having the following composition.

[0119] [Table 16]

[0120] Experimental Example 5: Evaluation of the stability of tamperacept ophthalmic compositions The stability of the tamperenacept ophthalmic composition of Preparation Example 1 was evaluated.

[0121] The composition of Preparation Example 1 was stored under long-term storage conditions of 5°C (with fine humidity adjustment) and accelerated storage conditions of 25°C / 60% RH, and the aggregate analysis using SEC-HPLC and charge variant analysis using IEX-HPLC were performed as described above.

[0122] [Table 17]

[0123] Table 17 and Figure 21 show the results of aggregate analysis using SEC-HPLC. As shown in Table 17 and Figure 21, the composition of Preparation Example 1 was confirmed to be stable, with aggregation occurring within 5% when stored for 3 years under long-term storage conditions at 5°C. Furthermore, it was confirmed that aggregation occurred within 5% when stored for 6 months under accelerated conditions at 25°C / 60% RH.

[0124] [Table 18]

[0125] Table 18 and Figures 22 to 23 show the results of charge variant analysis using IEX-HPLC. As shown in Table 18 and Figures 22 to 23, when the composition of Preparation Example 1 was stored for 3 years under long-term storage conditions at 5°C, acidic variants were induced within 10%, confirming that the composition was stable during the storage period. Basic variants were also induced within 10%, confirming that the composition was stable during the storage period.

[0126] Furthermore, when the strain was stored for six months under accelerated conditions of 25°C / 60% RH, acidic mutants were induced within 10%. Basic mutants were also induced within 10%, and the strain was stable during the storage period.

[0127] [Part B] Clinical trial of tamperacept ophthalmic composition The following clinical trial was conducted to evaluate the safety and efficacy of ophthalmic compositions containing tampercept versus placebo.

[0128] Clinical trial method This was a randomized, double-blind, placebo-controlled, multi-center clinical trial targeting patients with xerophthalmia. 637 patients were administered eye drops twice daily for eight weeks, and the primary evaluation indices were the Inferior Corneal Staining Score (ICSS) and Ocular Discomfort Score (ODS), with secondary evaluation indices being the Central Corneal Staining Score (CCSS), Total Corneal Staining Score (TCSS), and Eye Dryness Score (EDS).

[0129] 1. Clinical trial schedule The clinical trial was conducted according to the procedures shown in Figure 24. The subjects visited the clinical trial institution a total of six times during the 10-week clinical trial period, including a two-week screening process. Primary screening was conducted at the first visit, and secondary screening was conducted at the second visit. Patients who passed both screening processes were administered the drug according to the following administration method.

[0130] 2. Screening Method A total of 1,109 subjects were screened at 12 clinical trial sites, of which 472 dropped out during the screening process, leaving a total of 637 subjects receiving medication. During the screening period, subjects were exposed to a controlled CAE (Registered Trademark) chamber for 90 minutes twice (Visit 1 and Visit 2). The subject selection criteria were as follows:

[0131] Visit 1 Screening Criteria: 1) A history of dry eye syndrome for at least 6 months 2) People who have used or attempted to use artificial tears due to dry eyes within the past 6 months 3) Visual acuity 0.7 (logMAR standard) or less

[0132] Screening Criteria for Visits 1 and 2: 1) Ora Calibra® Ocular Discomfort & 4-Symptom Questionnaire score of 2 or more, 2) Schirmer score: 1mm or more and 10mm or less; 3) Corneal staining index of 2 or more points in at least one area; 4) Conjunctival redness score of 1 or more points, 5) After CAE exposure, the ICSS score must increase by 1 point or more, and the ODS score, measured every 5 minutes for 90 minutes of CAE exposure, must show a score of 3 points or more at two or more consecutive points. 2)~5) must be from the same eyeball.

[0133] Subjects who were finalized at Visit 2 were randomly assigned to receive the study treatment in a double-blind fashion for 8 weeks.

[0134] 3.Patient group A total of 637 subjects who passed the screening process were enrolled in the clinical trial and were randomly assigned to receive the study treatment in a double-blind manner. 318 patients received the study drug, tamperacept ophthalmic composition (0.25%), and 319 patients received a placebo. Demographic information for the clinical trial subjects is shown in Table 19 below.

[0135] [Table 19]

[0136] 4. Test drug and placebo The test drug used was the tamperacept ophthalmic composition (0.25%) prepared in Preparation Example 1. The placebo was prepared with the same composition as the test drug except that it did not contain tamperacept.

[0137] 5. Medication method and evaluation [Table 20]

[0138] Starting with the second visit, subjects self-administered the study drug or placebo twice daily (BID) for 8 weeks and completed daily symptom assessments, as shown in Table 20. Patients visited the clinical trial site on Days 1 (Visit 2), 8 (Visit 3), 15 (Visit 4), 29 (Visit 5), and 57 (Visit 6) after starting dosing. Signs and symptoms were assessed for these patients. Patients were exposed to the CAE® chamber on Days 15 (Visit 4), 29 (Visit 5), and 57 (Visit 6) after starting dosing. Signs and symptoms of xerophthalmia were assessed before, during, and after exposure. Signs and symptoms were assessed only on Day 8 (Visit 3) after starting dosing, without exposure to the CAE® chamber.

[0139] 6. Objective sign evaluation index The Corneal Staining Score (CSS) was used as an objective indicator for evaluating signs of dry eye. The CSS method involves dropping a stain into the eye, checking the degree of corneal damage, and assigning a score to the condition.

[0140] In this clinical trial, the Ora Calibra® Corneal Staining Index was used. Figure 25 illustrates the method for assessing the corneal staining index. As shown in Figure 25, the cornea was divided into the central (black), upper (light gray), and lower (dark gray) regions, and the degree of staining in each region was evaluated in 0.5-point increments from 0 to 4 points according to the criteria in Table 21 (the higher the score, the more severe the corneal damage). The evaluation results were displayed as CCSS for the central region, SCSS for the upper region, and ICSS for the lower region, and these were all summed to form an index called TCSS.

[0141] [Table 21]

[0142] 7. Subjective Symptom Assessment Index The symptom index for dry eye syndrome is a questionnaire that measures the degree of improvement in the patient's disease symptoms subjectively, and is evaluated using ODS, EDS, etc. depending on the type and format of the questionnaire.

[0143] 7-1.Ocular discomfort index (ODS) In this clinical trial, the Ora Calibra® Ocular Discomfort Scale (ODS) was used to assess ODS.

[0144] The degree of ocular discomfort felt by the patient is evaluated and graded individually for each eye using a score of 0 to 4 points according to the criteria in Table 22 below (the higher the score, the greater the ocular discomfort).

[0145] [Table 22]

[0146] 7-2.Dry Eye Sensation Index (EDS) The dry eye sensitivity index is an index of the dry eye sensation item on the visual analog scale (VAS). Subjects rate the dry eye sensation index using a number between 0 and 100, with 0 corresponding to "no dryness" and 100 corresponding to "extreme dryness."

[0147] 8. Schirmer test The Schirmer Tear Test (STT) was used as another method for assessing dry eye syndrome. The Schirmer Tear Test (STT) involves placing a Schirmer test strip on the edge of the lower eyelid of each patient's eye, closing the eyes for 5 minutes, then removing the Schirmer strip and measuring and recording the length (mm) of the moist area.

[0148] Clinical trial results 1. Baseline evaluation As discussed above in the clinical trial method, the objective corneal staining index and the subjective symptoms of the dry eye syndrome (EDS) and ocular discomfort index (ODS) were assessed on the first day of the clinical trial before the subjects were exposed to the CAE® chamber. The four corneal staining indices (ICSS, CCSS, SCSS, TCSS), the dry eye syndrome (EDS), and the ocular discomfort index (ODS) for the 318 patients administered the test drug and the 319 patients administered the placebo are shown in Table 23 below.

[0149] [Table 23]

[0150] 2. Corneal staining index evaluation results After 8 weeks of clinical drug administration, the change in the corneal staining index from baseline was confirmed as an objective sign for randomly assigned subjects (Intent-to-Treat (ITT) Population). Figure 26 shows the change in the values ​​of the inferior, superior, and central cornea, as well as the sum of these values ​​for the entire cornea (Total), from baseline to the end of administration (Week 8) for randomly assigned subjects (ITT Population, Observed Data Only).

[0151] As shown in Figure 26, subjects administered the test drug demonstrated significant improvement in corneal damage compared to placebo at week 8 in the central (CCSS, p=0.024) and total corneal (TCSS, p=0.045) areas.

[0152] Table 24 and Figures 27 to 28 show the changes and statistical analysis results in the CCSS and TCSS indices for all subjects over the 8-week period.

[0153] [Table 24] Covariances and significance levels were calculated using treatment, baseline score, and clinical trial site as covariates.

[0154] 3. Ocular Discomfort Index (ODS) and Eye Dryness Index (EDS) Evaluation Results After 8 weeks of clinical drug administration, the changes in subjective symptoms, ODS and EDS, from the baseline were confirmed.

[0155] Table 25 shows the mean change in EDS from baseline at week 8 for all subjects and for subjects with or without experience of using artificial tears within one month of Visit 1, and Figure 29 shows the mean change in EDS from baseline at week 8 for subjects who had experience of using artificial tears within one month of Visit 1.

[0156] As shown in Table 25 and Figure 29, the test drug did not show a significant improvement in EDS, a subjective indicator of dry eye syndrome, compared to placebo in randomly assigned subjects (ITT Population, Observed Data Only). However, a significant effect was confirmed in a relatively symptom-sensitive patient group who had used artificial tears within 30 days of Visit 1 (Day -14) of the clinical trial (p=0.0334).

[0157] The Ocular Discomfort Index (ODS) showed an effect in all subjects at 2 and 4 weeks.

[0158] Regarding symptoms, which are assessed by questionnaires on patients' subjective sensations, no significant difference was confirmed in the ODS, which was the primary evaluation variable, in the ITT analysis. However, when a subgroup analysis was conducted targeting only patients who had used artificial tears within one month of Visit 1 and were relatively sensitive to symptoms, a significant difference was confirmed in the EDS.

[0159] [Table 25]

[0160] 4. Clinical trial results for patient subgroups Patients enrolled in the clinical trial were subgrouped according to baseline severity, i.e., the severity of their disease at the start of the clinical trial, and signs and symptoms-related indicators were analyzed.

[0161] Table 26 and Figure 30 summarize the mean change in CCSS according to disease severity at the 8-week baseline.

[0162] [Table 26]

[0163] As shown in Table 26 and Figure 30, it was confirmed that the efficacy of the test drug compared to the placebo was more pronounced in the patient group with a high baseline severity.

[0164] Analysis of all subjects enrolled in the clinical trial showed differences from placebo in CCSS indices from one week after starting study medication, which persisted through week 8 (p=0.0239, two-sample t-test).

[0165] Of all patients, only those with severe corneal damage, with a baseline TCSS score of 5 or more, were analyzed, and a significant improvement in CCSS after 8 weeks of treatment was confirmed compared to placebo (p=0.0031).

[0166] The significance (p=0.0001) was confirmed in the analysis of only patients who had central corneal damage, i.e., a baseline CCSS score of 2 or more, at the start of treatment.

[0167] Furthermore, the significance was confirmed when only patients with a baseline Schirmer test score between 1 and 7, i.e., patients with relatively low tear secretion, were analyzed (p=0.0008).

[0168] In conclusion, the efficacy of the study drug compared with placebo was more pronounced in patients with higher baseline severity of signs, and this was also observed in patients with higher symptom baseline severity or combined sign and symptom baseline severity.

[0169] Furthermore, in an analysis of all patients, the test drug also showed a significant improvement in corneal damage compared to placebo at week 8 in the overall corneal staining index, which is the sum of the three corneal regions (CCSS, ICSS, SCSS) - that is, the TCSS index (p=0.0452, ANCOVA).The therapeutic effect of the test drug on the TCSS index was confirmed to be more evident in the analysis of the patient group with a high baseline severity among all patients.

[0170] Table 27 below and Figure 31 show the mean change in TCSS according to disease severity from baseline to 8 weeks.

[0171] Of all subjects, only patients with a baseline TCSS score of 5 or higher, i.e., patients with severe corneal damage, were included in the analysis. Results confirmed that the improvement in TCSS after 8 weeks of treatment (p=0.0109) was significant compared to placebo.

[0172] Significance was also confirmed in the results of an analysis of only patients whose central corneal damage level, i.e., whose CCSS score at the baseline was 2 or more at the time of initiation of treatment (p=0.0019).

[0173] The significance was also confirmed in the results of an analysis of only patients whose STT at baseline was between 1 and 7, i.e., patients with relatively low tear secretion (p=0.0025).

[0174] Similar findings were also observed in patients with higher symptom criteria or higher sign and symptom criteria, and were confirmed by two-sample t-test statistical analysis.

[0175] [Table 27]

Claims

1. An ophthalmic composition comprising tampanacept as an active ingredient for treating xerophthalmia in patients with moderate to severe dry eye.

2. The ophthalmic composition of claim 1, wherein the patient has a corneal staining index of 2 or more in at least one area of ​​the inferior cornea, the central cornea, and the superior cornea, and a Schirmer test score of 1 or more and 7 or less.

3. 10. The ophthalmic composition of claim 1, wherein improvement in moderate to severe dry eye symptoms begins to appear within 8 weeks of administration of the ophthalmic composition.

4. 4. The ophthalmic composition of claim 3, wherein the improvement in symptoms is measured by a decrease in total corneal staining score (TCSS) and central corneal staining score (CCSS).

5. 10. The ophthalmic composition of claim 1, wherein the improvement in the moderate to severe dry eye symptoms begins to appear within 8 weeks of administration of the ophthalmic composition.

6. The ophthalmic composition of claim 5, wherein the improvement in symptoms is measured by a decrease in the dry eye symptom score (EDS).

7. 10. The ophthalmic composition of claim 1, wherein the patient has a central corneal staining score (CCSS) of 2 or greater before administration of the ophthalmic composition.

8. 10. The ophthalmic composition of claim 1, wherein the patient has a Total Corneal Staining Score (TCSS) of 5 or greater prior to administration of the ophthalmic composition.

9. The ophthalmic composition according to claim 1 , wherein the patient has a Schirmer test score of 1 or more and 7 or less before administration of the ophthalmic composition.

10. The ophthalmic composition according to claim 1 , wherein the patient has a Schirmer test score of 1 or more and 3 or less before administration of the ophthalmic composition.

11. The ophthalmic composition of claim 1 , wherein the patient has an ocular discomfort score (ODS) of 3 or greater before administration of the ophthalmic composition.

12. 10. The ophthalmic composition of claim 1, wherein the patient has a Total Corneal Staining Score (TCSS) of 5 or greater and an Ocular Discomfort Score (ODS) of 3 or greater prior to administration of the ophthalmic composition.

13. 2. The ophthalmic composition of claim 1, wherein the patient has a Total Corneal Staining Score (TCSS) of 4 or greater and an Eye Dryness Score (EDS) of 40 or greater prior to administration of the ophthalmic composition.

14. The ophthalmic composition of claim 1 , wherein the ophthalmic composition is administered at least once daily in the form of eye drops.

15. The ophthalmic composition of claim 1 , wherein the ophthalmic composition is administered twice daily in the form of eye drops.

16. 10. The ophthalmic composition of claim 1, wherein the ophthalmic composition comprises tamperenacept and a buffer system having a pH of 5.0 to 6.5, and the ophthalmic composition is substantially free of a stabilizer.

17. The ophthalmic composition of claim 1, wherein the ophthalmic composition comprises 0.01 to 1% (w / v) of tamperacept.

18. A method for treating dry eye, comprising administering to a patient having moderate to severe dry eye an ophthalmic composition containing tampercept as an active ingredient.

19. 19. The method for treating dry eye disease according to claim 18, wherein the patient has a corneal staining index of 2 or more in at least one region of the inferior cornea, the central cornea, and the superior cornea, and a Schirmer test score of 1 or more and 7 or less.

20. 20. The method of claim 18, wherein improvement in moderate to severe dry eye signs begins to appear within 8 weeks of administration of the ophthalmic composition.

21. 21. The method of treating dry eye disease of claim 20, wherein the improvement in symptoms is measured by a decrease in total corneal staining score (TCSS) and central corneal staining score (CCSS).

22. 20. The method of claim 18, wherein the improvement in moderate to severe dry eye symptoms begins to appear within 8 weeks of administration of the ophthalmic composition.

23. 23. The method of claim 22, wherein the improvement in symptoms is measured by a decrease in the eye dryness score (EDS).

24. 20. The method of claim 18, wherein the patient has a central corneal staining score (CCSS) of 2 or greater before administration of the ophthalmic composition.

25. 20. The method of treating dry eye disease of claim 18, wherein the patient has a Total Corneal Staining Scale (TCSS) of 5 or greater prior to administration of the ophthalmic composition.

26. The method for treating dry eye disease according to claim 18, wherein the patient has a Schirmer test score of 1 or more and 7 or less before administration of the ophthalmic composition.

27. The method for treating dry eye disease according to claim 18, wherein the patient has a Schirmer test score of 1 or more and 7 or less before administration of the ophthalmic composition.

28. 19. The method of treating dry eye disease according to claim 18, wherein the patient has an ocular discomfort score (ODS) of 3 or greater before administration of the ophthalmic composition.

29. 20. The method of claim 18, wherein the patient has a Total Corneal Staining Score (TCSS) of 5 or greater and an Ocular Discomfort Score (ODS) of 3 or greater prior to administration of the ophthalmic composition.

30. 20. The method of claim 18, wherein the patient has a Total Corneal Staining Score (TCSS) of 4 or greater and an Eye Dryness Score (EDS) of 40 or greater prior to administration of the ophthalmic composition.

31. 20. The method for treating dry eye disease according to claim 18, wherein the ophthalmic composition is administered in the form of eye drops at least once a day.

32. 20. The method of treating dry eye disease of claim 18, wherein the ophthalmic composition is administered twice daily in the form of eye drops.

33. 20. The method of treating dry eye disease of claim 18, wherein the ophthalmic composition comprises tamperenacept and a buffer system having a pH of 5.0 to 6.5, and the ophthalmic composition is substantially free of a stabilizer.

34. 19. The method of treating dry eye disease according to claim 18, wherein the ophthalmic composition comprises 0.01% (w / v) to 1% (w / v) of tamperacept.

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