Formulation for the treatment of dry eye disease

Abstract

The present disclosure relates to an ophthalmic formulation of a lipidated kemptin composition comprising, inter alia, (a) kemptin or a fragment or analog thereof, and (b) a lipid entity linked to kemptin or a fragment or analog thereof, which is stable in terms of pH, weight osmolality, physical appearance, and purity of the kemptin composition when stored under normal storage conditions. One aspect of the present disclosure also relates to the use of the aforementioned ophthalmic formulation for treating inflammatory conditions including, but not limited to, ocular inflammation, dry eye disease, and neuropathic pain of the eye.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Patent Application No. 63 / 341,205, filed May 12, 2022, and U.S. Provisional Patent Application No. 63 / 434,042, filed December 20, 2022, the contents of each of which are incorporated herein by reference in their entirety.

[0002] Incorporation by reference of sequence listing The contents of the electronic sequence listing (OKYO_008_001WO_SeqList_ST26.xml; size: 6,273 bytes; and creation date: April 19, 2023) are incorporated herein by reference in their entirety. [Background technology]

[0003] There are various inflammatory conditions that affect the eye, including, but not limited to, ocular inflammation, dry eye disease (DED), and ocular neuropathic pain. Ocular inflammation can be caused by microbial infection of the eye. Such infections can be fungal, viral, or bacterial. Ocular inflammation can also be caused by trauma, burns, autoimmune diseases, medications, contact lenses, or other external stimuli. Neuropathic pain is a major health problem affecting approximately 7% of the general population. Up to 50% of patients fail standard treatments.

[0004] DED is a multifactorial disorder of the tears and ocular surface, with inflammation contributing to its pathogenesis. Dry eye is often a common, long-term problem, especially in older adults. In 2000, its prevalence in the United States was estimated at approximately 17% in women and 12% in men, but has increased in recent years and is estimated to exceed 50%. People with dry eye either do not produce enough tears or have poor-quality tears. Tears are produced by several glands in and around the eyelids. Tear production tends to decrease with age, with various medical conditions, or as a side effect of certain medications. Environmental conditions such as wind and dry weather can also reduce tear production by increasing tear evaporation. If the normal amount of tear production decreases or tears evaporate too rapidly from the eye, dry eye symptoms can develop. Regarding tear quality, tears are composed of three layers: oil, water, and mucus. Each component protects and nourishes the anterior surface of the eye. The smooth oil layer helps prevent evaporation of the aqueous layer, and the mucin layer spreads the tears evenly over the surface of the eye. If any of the three tear layers is deficient, causing tears to evaporate too quickly or not spread evenly over the cornea, dry eye symptoms can develop. A common form of dry eye occurs when the aqueous layer of tears is insufficient. This condition is also called keratoconjunctivitis sicca (KCS).

[0005] Current treatments for dry eye are symptomatic, focusing on tear replacement to alleviate symptoms. Five approved products are available in the United States: Restasis®, Xiidra®, CEQUA™, Eysuvis™, and TYRVAYA™. Restasis® (cyclosporine ophthalmic emulsion, 0.05%), a calcineurin inhibitor immunosuppressant, is a topical immunomodulator with anti-inflammatory effects. While approved for use in the treatment of DED in the United States, most of these products cause adverse side effects, including ocular irritation, taste abnormalities, instillation site pain, and conjunctival hyperemia.

[0006] OK-101 is a lipidated chemerin peptide designed to bind with high affinity to the ChemR23 receptor. Binding of OK-101 to ChemR23 has been shown to result in anti-inflammatory activity in a mouse model of DED. OK-101 was developed using membrane-anchored peptide (MAP) technology to create a novel, long-acting drug candidate for treating DED. OK-101 was also designed to enhance its residence time in the ocular environment by including a lipid "anchor" within its molecular structure, thereby counteracting shedding. OK-101 has also been shown to resolve inflammation in animal models of asthma and to modulate the inflammatory environment in autoimmune diseases by recruiting regulatory T cells (Tregs) and attenuating neuropathic pain in mice (Stevenson, Chauhan et al. 2012; Doyle, Krishnaji et al. 2014). OKYO's studies demonstrated a significant reduction in corneal permeability with topical application of OK-101 (0.04%) compared with vehicle in an experimental model of dry eye disease in mice. Furthermore, OK-101 normalized goblet cell density, reduced the number of CD4+ T cells (a biomarker of inflammation), and increased regulatory T cells in the draining lymph nodes of OK-101-treated mice compared with vehicle in a dry eye mouse model. Furthermore, in a separate set of animal model experiments, OK-101 was shown to exhibit potent ocular pain-reducing activity in a mouse model of corneal neuropathic pain. Due to its potential anti-inflammatory and analgesic properties, OK-101 is currently being developed by OKYO Pharma US, Inc. for the treatment of dry eye disease.

[0007] There is a need for therapeutic formulations for the treatment of ocular inflammatory conditions, such as DED, that have fewer or no adverse effects and are well tolerated by the patient's eye. To this end, the stability of such ophthalmic formulations with respect to variability in pH, osmolality, etc. is important.

[0008] The present disclosure addresses this need for patients suffering from various inflammatory conditions, including but not limited to ocular inflammation, DED, and ocular neuropathic pain, by providing an ophthalmic formulation of OK-101 that is similar in composition to human tears and stable in terms of pH, osmolality, physical appearance, and purity for at least 1-6 months. Summary of the Invention

[0009] The present disclosure provides an ophthalmic formulation comprising: (a) about 0.1% to about 0.5% w / v NaCl; (b) about 25 mM to about 100 mM phosphate buffered saline; and (c) a chemerin fragment consisting of the sequence YFPGQFAFS (SEQ ID NO: 2) or YFPGQFAFS (SEQ ID NO: 2). * -FLPS * -QFA * -Tic-S (SEQ ID NO: 3), * represents a D-amino acid, Tic represents 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, and the chemerin fragment or chemerin analog is linked to a lipid entity via a linker; and a lipidated chemerin composition, the lipidated chemerin composition having a pH of about 6.5 to about 8.5 and an osmolality of about 200 to about 450 mOsm / kg.

[0010] The present disclosure also provides a method of treating an inflammatory condition in a subject in need thereof, comprising topically administering to the eye of the subject a therapeutically effective amount of a formulation disclosed herein.

[0011] The present disclosure also provides a method of treating pain in a subject in need thereof, comprising topically administering to the eye of the subject a therapeutically effective amount of a formulation disclosed herein.

[0012] The present disclosure also provides kits for administering the ophthalmic formulations disclosed herein to a subject in need thereof. Methods of making the ophthalmic formulations disclosed herein are also provided.

[0013] Any aspect or embodiment described herein can be combined with any other aspect or embodiment disclosed herein. While the present disclosure has been described in conjunction with the detailed description, the foregoing description is intended to be illustrative and not limiting of the scope of the disclosure, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0014] The patent and scientific literature referred to herein establishes knowledge that is available to those skilled in the art. All U.S. patents and published or unpublished U.S. patent applications cited herein are incorporated by reference. All published foreign patents and patent applications cited herein are incorporated by reference. All other published references, documents, manuscripts, and scientific literature cited herein are incorporated by reference. [Brief explanation of the drawings]

[0015] [Figure 1A] 1A and 1B are charts showing the pH and solubility changes of a lipidated Chemerin composition (OK-101) of the present disclosure in 100 mM sodium phosphate buffer (FIG. 1A) and 50 mM sodium phosphate buffer (FIG. 1B). The pH values and concentrations of OK-101 achieved are shown. [Figure 1B] 1A and 1B are charts showing the pH and solubility changes of a lipidated Chemerin composition (OK-101) of the present disclosure in 100 mM sodium phosphate buffer (FIG. 1A) and 50 mM sodium phosphate buffer (FIG. 1B). The pH values and concentrations of OK-101 achieved are shown.

[0016] [Figure 2A]The precision and detection limit of the HPLC assay used to detect OK-101 are shown. Figure 2A shows the concentration of OK-101 (mg / mL) plotted against the detection signal peak area. Figure 2B shows the detection signal peak area of replicate injections (input) of the first standard in Figure 2A to demonstrate the consistency and precision of detection. Figure 2C is a standard curve representing Figure 2A. Figure 2D shows various 0.2 μm membranes tested and considered for use in the HPLC assay. [Figure 2B] The precision and detection limit of the HPLC assay used to detect OK-101 are shown. Figure 2A shows the concentration of OK-101 (mg / mL) plotted against the detection signal peak area. Figure 2B shows the detection signal peak area of replicate injections (input) of the first standard in Figure 2A to demonstrate the consistency and precision of detection. Figure 2C is a standard curve representing Figure 2A. Figure 2D shows various 0.2 μm membranes tested and considered for use in the HPLC assay. [Figure 2C] The precision and detection limit of the HPLC assay used to detect OK-101 are shown. Figure 2A shows the concentration of OK-101 (mg / mL) plotted against the detection signal peak area. Figure 2B shows the detection signal peak area of replicate injections (input) of the first standard in Figure 2A to demonstrate the consistency and precision of detection. Figure 2C is a standard curve representing Figure 2A. Figure 2D shows various 0.2 μm membranes tested and considered for use in the HPLC assay. [Figure 2D] The precision and detection limit of the HPLC assay used to detect OK-101 are shown. Figure 2A shows the concentration of OK-101 (mg / mL) plotted against the detection signal peak area. Figure 2B shows the detection signal peak area of replicate injections (input) of the first standard in Figure 2A to demonstrate the consistency and precision of detection. Figure 2C is a standard curve representing Figure 2A. Figure 2D shows various 0.2 μm membranes tested and considered for use in the HPLC assay.

[0017] [Figure 3]1 is a chart showing the conditions and schedule for testing formulations containing 0.05% OK-101 product in 100 mM sodium phosphate buffer with 0.3% NaCl and 0.05% OK-101 product in 100 mM sodium phosphate buffer with 1.4% mannitol. Specific tests are shown as follows: A, tests performed on A include appearance, pH, osmolality, color, assay / potency, and impurities by HPLC; S includes storage only.

[0018] [Figure 4A] Figure 4A shows the changes in appearance, pH, osmolality, and purity of formulations of OK-101 product over a one-month storage period. Figure 4A is a photograph showing the clear, colorless appearance of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH. Figure 4B is a chart showing the changes in pH, osmolality, % assay purity, and % area for each formulation and storage condition corresponding to the respective relative retention times (RRTs) of T=0 and T=1 month for each formulation and storage condition. Figures 4C, 4D, and 4E are charts showing the purity of OK-101 product and formulations as determined using the HPLC assay at T = 0 months, T = 1 month under storage conditions of 40°C / 75%RH, and T = 1 month under storage conditions of 25°C / 60%RH. The % assay purity levels and % area values for each formulation and storage condition corresponding to each relative retention time (RRT) are shown. [Figure 4B]Figure 4A shows the changes in appearance, pH, osmolality, and purity of formulations of OK-101 product over a one-month storage period. Figure 4A is a photograph showing the clear, colorless appearance of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH. Figure 4B is a chart showing the changes in pH, osmolality, % assay purity, and % area for each formulation and storage condition corresponding to the respective relative retention times (RRTs) of T=0 and T=1 month for each formulation and storage condition. Figures 4C, 4D, and 4E are charts showing the purity of OK-101 product and formulations as determined using the HPLC assay at T = 0 months, T = 1 month under storage conditions of 40°C / 75%RH, and T = 1 month under storage conditions of 25°C / 60%RH. The % assay purity levels and % area values for each formulation and storage condition corresponding to each relative retention time (RRT) are shown. [Figure 4C] Figure 4A shows the changes in appearance, pH, osmolality, and purity of formulations of OK-101 product over a one-month storage period. Figure 4A is a photograph showing the clear, colorless appearance of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH. Figure 4B is a chart showing the changes in pH, osmolality, % assay purity, and % area for each formulation and storage condition corresponding to the respective relative retention times (RRTs) of T=0 and T=1 month for each formulation and storage condition. Figures 4C, 4D, and 4E are charts showing the purity of OK-101 product and formulations as determined using the HPLC assay at T = 0 months, T = 1 month under storage conditions of 40°C / 75%RH, and T = 1 month under storage conditions of 25°C / 60%RH. The % assay purity levels and % area values for each formulation and storage condition corresponding to each relative retention time (RRT) are shown. [Figure 4D]Figure 4A shows the changes in appearance, pH, osmolality, and purity of formulations of OK-101 product over a one-month storage period. Figure 4A is a photograph showing the clear, colorless appearance of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH. Figure 4B is a chart showing the changes in pH, osmolality, % assay purity, and % area for each formulation and storage condition corresponding to the respective relative retention times (RRTs) of T=0 and T=1 month for each formulation and storage condition. Figures 4C, 4D, and 4E are charts showing the purity of OK-101 product and formulations as determined using the HPLC assay at T = 0 months, T = 1 month under storage conditions of 40°C / 75%RH, and T = 1 month under storage conditions of 25°C / 60%RH. The % assay purity levels and % area values for each formulation and storage condition corresponding to each relative retention time (RRT) are shown. [Figure 4E] Figure 4A shows the changes in appearance, pH, osmolality, and purity of formulations of OK-101 product over a one-month storage period. Figure 4A is a photograph showing the clear, colorless appearance of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH. Figure 4B is a chart showing the changes in pH, osmolality, % assay purity, and % area for each formulation and storage condition corresponding to the respective relative retention times (RRTs) of T=0 and T=1 month for each formulation and storage condition. Figures 4C, 4D, and 4E are charts showing the purity of OK-101 product and formulations as determined using the HPLC assay at T = 0 months, T = 1 month under storage conditions of 40°C / 75%RH, and T = 1 month under storage conditions of 25°C / 60%RH. The % assay purity levels and % area values for each formulation and storage condition corresponding to each relative retention time (RRT) are shown.

[0019] [Figure 5A]Figure 5A shows the changes in appearance, pH, osmolality, and purity of formulations of 0.05% w / v OK-101 product over storage periods of 2, 3, and 6 months. Figure 5B shows the changes in appearance, pH, osmolality, and purity of formulations of 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 0.3% NaCl and 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for T = 2 months and T = 3 months. The pH values, osmolality values (mOsm), % assay purity levels, and % area values for each formulation and storage condition corresponding to each relevant retention time (RRT) for each formulation and storage condition are shown. Figure 5B is a chart showing the change in appearance, pH, osmolality, and % assay purity level of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for T = 6 months. Figure 5C is a photograph showing the clear and colorless appearance of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for 3 months. [Figure 5B]Figure 5A shows the changes in appearance, pH, osmolality, and purity of formulations of 0.05% w / v OK-101 product over storage periods of 2, 3, and 6 months. Figure 5B shows the changes in appearance, pH, osmolality, and purity of formulations of 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 0.3% NaCl and 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for T = 2 months and T = 3 months. The pH values, osmolality values (mOsm), % assay purity levels, and % area values for each formulation and storage condition corresponding to each relevant retention time (RRT) for each formulation and storage condition are shown. Figure 5B is a chart showing the change in appearance, pH, osmolality, and % assay purity level of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for T = 6 months. Figure 5C is a photograph showing the clear and colorless appearance of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for 3 months. [Figure 5C]Figure 5A shows the changes in appearance, pH, osmolality, and purity of formulations of 0.05% w / v OK-101 product over storage periods of 2, 3, and 6 months. Figure 5B shows the changes in appearance, pH, osmolality, and purity of formulations of 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 0.3% NaCl and 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for T = 2 months and T = 3 months. The pH values, osmolality values (mOsm), % assay purity levels, and % area values for each formulation and storage condition corresponding to each relevant retention time (RRT) for each formulation and storage condition are shown. Figure 5B is a chart showing the change in appearance, pH, osmolality, and % assay purity level of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for T = 6 months. Figure 5C is a photograph showing the clear and colorless appearance of a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 0.3% NaCl and a 0.05% OK-101 product formulation in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40°C / 75% RH and 25°C / 60% RH for 3 months.

[0020] [Figure 6A]Figure 6A shows the changes in appearance, pH, osmolality, and purity of formulations of 0.1% w / v OK-101 product over 1-month and 2-month storage periods. Figure 6A is a chart showing the conditions and schedule for testing formulations containing 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 0.3% NaCl and 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 1.4% mannitol. Specific tests are indicated as follows: A, tests performed on A include appearance, pH, osmolality, color, assay / potency, and impurities by HPLC; S includes storage only. 6B is a chart showing the change in appearance, pH, osmolality, and % assay purity level for a formulation of 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 0.3% NaCl and a formulation of 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40° C. / 75% RH and 25° C. / 60% RH for T=1 month and T=2 months. The pH values, osmolality values (mOsm), and % assay purity levels for each formulation and storage condition are shown. [Figure 6B]Figure 6A shows the changes in appearance, pH, osmolality, and purity of formulations of 0.1% w / v OK-101 product over 1-month and 2-month storage periods. Figure 6A is a chart showing the conditions and schedule for testing formulations containing 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 0.3% NaCl and 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 1.4% mannitol. Specific tests are indicated as follows: A, tests performed on A include appearance, pH, osmolality, color, assay / potency, and impurities by HPLC; S includes storage only. 6B is a chart showing the change in appearance, pH, osmolality, and % assay purity level for a formulation of 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 0.3% NaCl and a formulation of 0.05% OK-101 product in 100 mM sodium phosphate buffer containing 1.4% mannitol stored under storage conditions of 40° C. / 75% RH and 25° C. / 60% RH for T=1 month and T=2 months. The pH values, osmolality values (mOsm), and % assay purity levels for each formulation and storage condition are shown. DETAILED DESCRIPTION OF THE INVENTION

[0021] The present disclosure provides an ophthalmic formulation comprising: (a) about 0.1% to about 0.5% w / v NaCl; (b) about 25 mM to about 100 mM phosphate buffered saline; and (c) a chemerin fragment consisting of the sequence YFPGQFAFS (SEQ ID NO: 2) or YFPGQFAFS (SEQ ID NO: 2). * -FLPS * -QFA * -Tic-S (SEQ ID NO: 3), *represents a D-amino acid, Tic represents 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, and the chemerin fragment or chemerin analog is linked to a lipid entity via a linker; and a lipidated chemerin composition, the lipidated chemerin composition having a pH of about 6.5 to about 8.5 and an osmolality of about 200 to about 450 mOsm / kg.

[0022] In some embodiments, a formulation of the present disclosure comprises 0.1% to 0.5% w / v NaCl (e.g., 0.1% to 0.2%, 0.2% to 0.3%, 0.3% to 0.4%, or 0.4% to 0.5% w / v NaCl). In some embodiments, a formulation of the present disclosure comprises 0.1% to 0.3% w / v NaCl (e.g., 0.1% to 0.2% or 0.2% to 0.3% w / v NaCl). In some embodiments, a formulation of the present disclosure comprises 0.3% w / v NaCl.

[0023] In some embodiments, a formulation of the present disclosure comprises 0.05% to 0.1% w / v (e.g., 0.05% to 0.075%, 0.075% to 0.1% w / v) of a lipidated chemerin composition disclosed herein. In some embodiments, a formulation of the present disclosure comprises about 0.05% of a lipidated chemerin composition disclosed herein. In some embodiments, a formulation of the present disclosure comprises 0.05% of a lipidated chemerin composition disclosed herein. In some embodiments, a formulation of the present disclosure comprises about 0.1% of a lipidated chemerin composition disclosed herein. In some embodiments, a formulation of the present disclosure comprises 0.1% of a lipidated chemerin composition disclosed herein.

[0024] In some embodiments, a formulation of the present disclosure comprises 25 mM to 50 mM (e.g., 25 mM to 30 mM, 30 mM to 35 mM, 35 mM to 40 mM, 40 mM to 45 mM, or 45 mM to 50 mM) phosphate buffered saline. In some embodiments, a formulation of the present disclosure comprises 50 mM to 100 mM (e.g., 50 mM to 55 mM, 55 mM to 60 mM, 60 mM to 65 mM, 65 mM to 70 mM, 70 mM to 75 mM, 75 mM to 80 mM, 80 mM to 85 mM, 85 mM to 90 mM, 90 mM to 95 mM, or 95 mM to 100 mM) phosphate buffered saline. In some embodiments, a formulation of the present disclosure comprises 50 mM phosphate buffered saline. In some embodiments, the formulations of the present disclosure comprise 100 mM phosphate buffered saline.

[0025] In some embodiments, a formulation of the present disclosure has a pH of 6.5 to 8.5 (e.g., 6.5 to 7.0, 7.0 to 7.5, 7.5 to 8.0, or 8.0 to 8.5). In some embodiments, a formulation of the present disclosure has a pH of about 7.2 to about 7.4. In some embodiments, a formulation of the present disclosure has a pH of 7.3 to 7.4. In some embodiments, a formulation of the present disclosure has a pH of 7.4 to 7.6.

[0026] In some embodiments, the formulations of the present disclosure have an osmolality of 200-450 mOsm / kg (e.g., 200-250, 250-300, 300-350, 350-400, or 400-450 mOsm / kg). In some embodiments, the formulations have an osmolality of about 290 to about 320 mOsm / kg (e.g., 290-300, 300-310, 310-315, 315-320, 320-325, 325-330, 330-335, 335-340, 340-345, or 345-350 mOsm / kg). In some embodiments, a formulation of the present disclosure has an osmolality of 290-325 mOsm / kg (e.g., 290-295, 295-300, 300-305, 305-310, 310-315, 315-325 mOsm / kg). In some embodiments, a formulation of the present disclosure has an osmolality of 313-322 mOsm / kg (e.g., 313-315, 315-317, 317-319, or 319-322 mOsm / kg). In some embodiments, a formulation of the present disclosure has an osmolality of 314-319 mOsm / kg (e.g., 314-315, 315-316, 316-317, or 318-319 mOsm / kg).

[0027] In some embodiments, the pH of a formulation of the present disclosure changes by 1.5% or less (0% to 0.25%, 0.25% to 0.5%, 0.5% to 0.75%, 0.75% to 1%, 1% to 1.25%, or 1.25% to 1.5%) over a period of 1 month to 6 months (e.g., 1 month to 2 months, 2 months to 3 months, 3 months to 4 months, 4 months to 5 months, or 5 months to 6 months) when stored at 25° C. to 40° C. and 60% to 75% relative humidity.

[0028] In some embodiments, the pH of a formulation of the present disclosure changes by 1% to 1.5% over a one-month period when stored at 25°C and 60% relative humidity. In some embodiments, the pH of a formulation of the present disclosure changes by 1% to 1.5% over a two-month period when stored at 25°C and 60% relative humidity. In some embodiments, the pH of a formulation of the present disclosure changes by 1% to 1.5% over a three-month period when stored at 25°C and 60% relative humidity. In some embodiments, the pH of a formulation of the present disclosure changes by 1% to 1.5% over a six-month period when stored at 25°C and 60% relative humidity.

[0029] In some embodiments, the pH of a formulation of the present disclosure does not change over a period of 1 month when stored at 25° C. and 60% relative humidity. In some embodiments, the pH of a formulation of the present disclosure does not change over a period of 2 months when stored at 25° C. and 60% relative humidity. In some embodiments, the pH of a formulation of the present disclosure does not change over a period of 3 months when stored at 25° C. and 60% relative humidity. In some embodiments, the pH of a formulation of the present disclosure does not change over a period of 6 months when stored at 25° C. and 60% relative humidity.

[0030] In some embodiments, the pH of a formulation of the present disclosure changes by 1% to 1.5% over a one-month period when stored at 40°C and 75% relative humidity. In some embodiments, the pH of a formulation of the present disclosure changes by 1% to 1.5% over a two-month period when stored at 40°C and 75% relative humidity. In some embodiments, the pH of a formulation of the present disclosure changes by 1% to 1.5% over a three-month period when stored at 40°C and 75% relative humidity. In some embodiments, the pH of a formulation of the present disclosure changes by 1% to 1.5% over a six-month period when stored at 40°C and 75% relative humidity.

[0031] In some embodiments, the pH of a formulation of the present disclosure does not change over a period of 1 month when stored at 40° C. and 75% relative humidity. In some embodiments, the pH of a formulation of the present disclosure does not change over a period of 2 months when stored at 40° C. and 75% relative humidity. In some embodiments, the pH of a formulation of the present disclosure does not change over a period of 3 months when stored at 40° C. and 75% relative humidity. In some embodiments, the pH of a formulation of the present disclosure does not change over a period of 6 months when stored at 40° C. and 75% relative humidity.

[0032] In some embodiments, the osmolality of the formulation changes by about 8% or less over a period of 1 month to 6 months when stored at a relative humidity of 25° C. to 40° C. In some embodiments, the osmolality of the formulation changes by 0.3% to 10% (e.g., 0.3% to 0.5%, 0.5% to 1.0%, 1.0%) over a period of 1 month to 6 months (e.g., 1 month to 2 months, 2 months to 3 months, 3 months to 4 months, 4 months to 5 months, or 5 months to 6 months) when stored at 25° C. to 40° C. and a relative humidity of 60% to 75%. % to 1.5%, 1.5% to 2%, 2% to 2.5%, 2.5% to 3%, 3% to 3.5%, 3.5% to 4%, 4% to 4.5%, 4.5% to 5%, 5% to 5.5%, 5.5% to 6%, 6% to 6.5%, 6.5% to 7%, 7% to 7.5%, 7.5% to 8%, 8% to 8.5%, 8.5% to 9%, 9% to 9.5%, or 9.5% to 10%) change.

[0033] In some embodiments, the osmolality of the formulation changes by 2% or less (e.g., 0.1% to 0.25%, 0.25% to 0.3%, 0.3% to 0.5%, 0.5% to 0.75%, 0.75% to 1.0%, 1.0% to 1.25%, or 1.25% to 1.5%) over a period of 1 month to 6 months (e.g., when stored at 25° C. and 60% relative humidity). In some embodiments, the osmolality of the formulation changes by 0.25% to 2% (e.g., 0.25% to 0.3%, 0.3% to 0.5%, 0.5% to 0.75%, 0.75% to 1.0%, 1.0% to 1.25%, or 1.25% to 1.5%, 1.5% to 2%) over a period of 1 month to 6 months (e.g., when stored at 25° C. and 60% relative humidity). In some embodiments, the osmolality of the formulation changes by 0.25% to 2% over a period of 1 month when stored at 25° C. and 60% relative humidity. In some embodiments, the osmolality of the formulation changes by no more than 0.25% to 2% over a period of 2 months when stored at 25° C. and 60% relative humidity. In some embodiments, the osmolality of the formulation changes by 0.25% to 2% over a period of 3 months when stored at 25° C. and 60% relative humidity. In some embodiments, the osmolality of the formulation changes by 0.25% to 2% over a period of 6 months when stored at 25° C. and 60% relative humidity.

[0034] In some embodiments, the osmolality of a formulation of the present disclosure does not change over a period of 1 month when stored at 25°C and 60% relative humidity. In some embodiments, the osmolality of a formulation of the present disclosure does not change over a period of 2 months when stored at 25°C and 60% relative humidity. In some embodiments, the osmolality of a formulation of the present disclosure does not change over a period of 3 months when stored at 25°C and 60% relative humidity. In some embodiments, the osmolality of a formulation of the present disclosure does not change over a period of 6 months when stored at 25°C and 60% relative humidity.

[0035] In some embodiments, the osmolality of the formulation changes by 8% or less (e.g., 1% to 1.5%, 1.5% to 2%, 2% to 2.5%, 2.5% to 5%, or 5% to 5.5%, 5.5% to 6%, 6% to 6.5%, 6.5% to 7%, 7% to 7.5%, or 7.5% to 8%) over a period of 1 month to 6 months (e.g., 1 month to 2 months, 2 months to 3 months, 3 months to 4 months, 4 months to 5 months, or 5 months to 6 months) when stored at 40° C. and 75% relative humidity. In some embodiments, the osmolality of the formulation changes by 1% to 8% (e.g., 1% to 1.5%, 1.5% to 2%, 2% to 2.5%, 2.5% to 5%, or 5% to 5.5%, 5.5% to 6%, 6% to 6.5%, 6.5% to 7%, 7% to 7.5%, or 7.5% to 8%) over a period of 1 month to 6 months (e.g., 1 month to 2 months, 2 months to 3 months, 3 months to 4 months, 4 months to 5 months, or 5 months to 6 months) when stored at 40° C. and 75% relative humidity. In some embodiments, the osmolality of the formulation changes by 1% to 8% over a period of 1 month when stored at 40° C. and 75% relative humidity. In some embodiments, the osmolality of the formulation changes by 1% to 8% over a period of 2 months when stored at 40° C. and 75% relative humidity. In some embodiments, the osmolality of the formulation changes by 1% to 8% over a 3 month period when stored at 40° C. and 75% relative humidity. In some embodiments, the osmolality of the formulation changes by 1% to 8% over a 6 month period when stored at 40° C. and 75% relative humidity.

[0036] In some embodiments, the osmolality of a formulation of the present disclosure does not change over a period of 1 month when stored at 40°C and 75% relative humidity. In some embodiments, the osmolality of a formulation of the present disclosure does not change over a period of 2 months when stored at 40°C and 75% relative humidity. In some embodiments, the osmolality of a formulation of the present disclosure does not change over a period of 3 months when stored at 40°C and 75% relative humidity. In some embodiments, the osmolality of a formulation of the present disclosure does not change over a period of 6 months when stored at 40°C and 75% relative humidity.

[0037] In some embodiments, the purity of the lipidated chemerin composition used to make the formulation is >94.6% (e.g., 94.6% to 95%, 95% to 96%, 96% to 97%, 97% to 98%, 98% to 99%, or 99% to 100%) and the peptide content is >95.9% (e.g., 95.9% to 97%, 97% to 98%, 98% to 99%, or 99% to 100%) as determined by HPLC. In some embodiments, the purity of the lipidated chemerin composition used to make the formulation is 95% to 98% (e.g., 95% to 96%, 96% to 97%, or 97% to 98%).

[0038] In some embodiments, in lipidated Chemerin compositions used to make formulations of the present disclosure, Tic represents: [ka]

[0039] In some embodiments, any of a variety of lipid entities can be utilized in the lipidated chemerin compositions used to prepare the formulations of the present disclosure. According to various embodiments, the lipid entity can include an entity capable of inserting into a lipid bilayer (e.g., a cell membrane). In some embodiments, the lipid entity can be incorporated into lipid rafts within a lipid bilayer (e.g., a cell membrane).

[0040] In some embodiments, the lipid entity can comprise saturated or unsaturated fatty acids. The numbers in the lipid name are used to describe the fatty acid chain on the lipid. The numbers are generally expressed in the format (number of carbons in the fatty acid chain):(number of double bonds in the fatty acid chain), e.g., 16:0 is 16 carbons in the fatty acid chain with 0 double bonds. Saturated or unsaturated fatty acids can comprise at least 4 carbons, at least 5 carbons, at least 6 carbons, at least 7 carbons, at least 8 carbons, at least 9 carbons, at least 10 carbons, or at least 15 carbons in the fatty acid chain. In some embodiments, saturated or unsaturated fatty acids can comprise about 4 to 24 carbons in the fatty acid chain. The number of double bonds in the fatty acid chain can range from 0 to 10, e.g., 0 to 8, 0 to 6, 1 to 8, or 1 to 6. For example, the lipid entity can be C22:0, C22:1, C22:2, C22:3, C22:4, C22:5, C22:6, C20:0, C20:1, C20:2, C20:3, C20:4, C20:5, C20:6, C18:0, C18:1, C18:2, C18:3, C18:4, C18:5, C18:6, C10:0, C10:1, C10:2, C10:3, C10:4, etc.

[0041] For example, lipid entities include α-linolenic acid, γ-linolenic acid, stearidonic acid, eicosapentaenoic acid, docosahexaenoic acid, linoleic acid, dihomo-γ-linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, paulinic acid, oleic acid, elaidic acid, gondoic acid, erucic acid, nervonic acid, mead acid, myristic acid, palmitic acid, stearic acid, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), GM1 ganglioside, GM2 ganglioside, GM3 ganglioside, The lipid entity may be selected from the group consisting of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), glycosphingolipids, sphingolipids, phosphatidylinositol 4,5-bisphosphate (PIP2), ceramide, cholesterol, ergosterol, phytosterols, hopanoids, steroids, fluorinated-GM1, fluorinated-GM2, and fluorinated-GM3. In some embodiments, the lipid entity may be α-linolenic acid. In some embodiments, the lipid entity may be γ-linolenic acid. In some embodiments, the lipid entity may be palmitic acid. In some embodiments, the lipid entity may be vaccenic acid. In some embodiments, the lipid entity may be oleic acid. In some embodiments, the lipid entity may be elaidic acid.

[0042] The binding of lipid entities to polypeptides is called lipidation.In some embodiments, lipidation can include N-myristoylation.As used herein, "N-myristoylation" refers to the binding of myristate to N-terminal glycine.

[0043] In some embodiments, lipidation can include palmitoylation. As used herein, "palmitoylation" refers to the creation of a thioester bond of a long-chain fatty acid on one or more cysteine residues present in a peptide or protein.

[0044] In some embodiments, lipidation comprises GPI-anchoring. As used herein, "GPI-anchoring" refers to the attachment of glycosylphosphatidylinositol (GPI) to the C-terminus of a protein.

[0045] In some embodiments, lipidation comprises prenylation. As used herein, "prenylation" refers to the formation of a thioether bond of an isoprenoid lipid (e.g., farnesyl (C-15) or geranylgeranyl (C-20)) to a cysteine present in a peptide or protein. In some embodiments, lipidation comprises geranylation. In some embodiments, lipidation comprises geranylgeranylation. In some embodiments, lipidation comprises the association of a ligand entity with any compound that is soluble in a cell membrane (e.g., 10:1 with an equilibrium constant K > 10).

[0046] In some embodiments, lipidation may involve one or more of the following: attachment of diacylglycerol to the side chain of the N-terminal cysteine of the peptide or protein via a sulfur atom; attachment of O-octanoyl to serine or threonine of the peptide or protein; and attachment of S-archeol to cysteine of the peptide or protein. In some embodiments, lipidation may occur at, for example, any lysine, glutamic acid, aspartic acid, serine, threonine, cysteine, and / or tyrosine. In some embodiments in which the chemerin analog contains one or more ornithines, lipidation may occur at any ornithine.

[0047] In some embodiments, the lipid entity may be linked to or near the N-terminus of Chemerin or a fragment or analog thereof, hi some embodiments, the lipid entity may be linked to or near the C-terminus of Chemerin or a fragment or analog thereof.

[0048] In some embodiments, lipidation can comprise fluorination.Fluorination can comprise the addition of one or more C6F13 chains.Without being bound by theory, it is believed that the presence of one or more C6F13 chains can allow lipid entities to separate from hydrocarbon lipid membrane components (see J.Am.Chem.Soc.2007,129,9037-9043; J.Phys.Chem.B,2008,112,8250-8256; J.Am.Chem.Soc.,2009,131,12091-12093).

[0049] In some embodiments, the presence of at least one alkene in the structure of a lipid entity increases its fluidity within a membrane (i.e., its ability to move within a membrane) compared to a similar lipid entity lacking at least one alkene. In some embodiments, lipid entities with increased fluidity may exhibit enhanced activity against targets (e.g., receptors, ion channels, or enzymes) with low density within a membrane. Without being bound by theory, lipid entities with increased ability to move within a membrane may encounter low-density targets more quickly than lipid entities with lower mobility within a membrane.

[0050] In some embodiments, the lipidated chemerin compositions used to make the formulations of the present disclosure can optionally include a linker connecting the lipid entity to the chemerin or a fragment or analog thereof. For example, the linker can have a length of from about 2 Å to about 175 Å. In some embodiments, the linker is from 30 Å to about 150 Å.

[0051] In some embodiments, the linker can comprise a peptide. In some embodiments, the peptide linker is about 2-20 amino acid residues in length. In some embodiments, the peptide linker is about 5-10 amino acid residues in length. According to various embodiments, the peptide linker can be designed to form one or more α-helices between the chemerin or fragment or analog thereof and the lipid entity. In some embodiments, the peptide linker can include multiple α-helices. In some embodiments, the multiple α-helices are contiguous. In some embodiments, the multiple α-helices are 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or more α-helices.

[0052] In some embodiments, the peptide linker can include a repeating unit, for example, multiple repeating glycine-asparagine (GN) units. In some embodiments, the peptide linker can include an epitope tag (e.g., a c-Myc tag) or other marker, allowing for identification and / or characterization of the provided agents and their fate in vitro and / or in vivo.

[0053] In some embodiments, the linker can include a non-peptide entity. In some embodiments, the non-peptide linker can be a synthetic polymer. According to various embodiments, the synthetic polymer can be any of a variety of lengths. In some embodiments, the linker including a synthetic polymer includes a monomeric unit of the polymer. In some embodiments, the linker including a synthetic polymer includes two or more monomeric units of the synthetic polymer (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 100, or more monomeric units).

[0054] In some embodiments, the linker can comprise at least one molecule of polyethylene glycol (PEG). Specific non-limiting examples of suitable polymer linkers include linkers having one or more monomer units according to one of the following formulas: [ka] wherein n represents an integer greater than or equal to 1. In some embodiments, n is an integer greater than or equal to 2 and less than or equal to 50, greater than or equal to 4 and less than or equal to 24, and / or greater than or equal to 8 and less than or equal to 24.

[0055] In some embodiments, in lipidated Chemerin compositions used to make the formulations of the present disclosure, the linker is selected from the group consisting of: [ka]

[0056] In some embodiments, the linker comprises polyethylene glycol, GG, KGG, or a combination thereof.

[0057] In some embodiments, the lipid entity is linked to or near the N-terminus of the chemerin fragment or chemerin analog. In some embodiments, the lipid entity is linked to or near the C-terminus of the chemerin fragment or chemerin analog.

[0058] In some embodiments, the linker can include 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC), benzophenone-4-isothiocyanate, bis-((N-iodoacetyl)piperazinyl)sulfonerhodamine, succinimidyl 2-(2-pyridyldithio)propionate (SPDP), 4-azido-2,3,5,6-tetrafluorobenzoic acid (ATFB), (N-((2-pyridyldithio)ethyl)-4-azidosalicylamido), succinimidyl trans-4-(maleimidylmethyl)cyclohexane-1-carboxylate (SMCC), and / or N-(t-BOC)-aminooxyacetic acid. Those skilled in the art can identify additional candidate linkers according to known methods.

[0059] In some embodiments, the linker can include both peptide and non-peptide entities.

[0060] In some embodiments, the linker is formed, at least in part, as a result of a Click reaction, further described below. In some embodiments, the Click reaction is an azide-alkyne Huisgen cycloaddition reaction.

[0061] Further examples of lipid entities, linkers, and methods of lipidation can be found in U.S. Patent Application Publication No. 20160052982, the contents of which are incorporated herein by reference.

[0062] In some embodiments, the lipidated Chemerin composition has the following structure: [ka]

[0063] In some embodiments, the formulations of the present disclosure are formulated for topical administration as eye drops.

[0064] CMKLR1 is a G protein-coupled receptor that has been shown to regulate nociception. This receptor is expressed in glia, dorsal root ganglion neurons, and immune cells. The endogenous ligand (agonist) of CMKLR1 is a 163-amino acid protein, chemerin. Chemerin, also known as retinoic acid receptor response protein 2 (RARRES2), tazarotene-inducible gene 2 protein (TIG2), or RAR-responsive protein TIG2, is a protein encoded by the RARRES2 gene in humans. The amino acid sequence of chemerin (Homo sapiens) is shown below in SEQ ID NO: 1.

[0065] NCBI reference sequence: NP_002880.1 MRRLLIPLAL WLGAVGVGVA ELTEAQRRGL QVALEEFHKH PPVQWAFQET SVESAVDTPF PAGIFVRLEF KLQQTSCRKR DWKKPECKVR PNGRKRKCLA CIKLGSEDKV LGRLVHCPIE TQVLREAEEH QETQCLRVQR AGEDPHSFYF PGQFAFSKAL PRS (SEQ ID NO: 1).

[0066] Chemerin is inactive as preprochemerin (having SEQ ID NO: 1) and is activated via C- and N-terminal cleavage to form a chemerin fragment having the amino acid sequence of positions 21 to 157 of SEQ ID NO: 1, which can function as an agonist of CMKLR1. This chemerin fragment has the following amino acid sequence: ELTEAQRRGL QVALEEFHKH PPVQWAFQET SVESAVDTPF PAGIFVRLEF KLQQTSCRKR DWKKPECKVR PNGRKRKCLA CIKLGSEDKV LGRLVHCPIE TQVLREAEEH QETQCLRVQR AGEDPHSFYF PGQFAFS (SEQ ID NO: 4).

[0067] In one aspect, the present disclosure provides a composition comprising (a) a chemerin or a fragment or analog thereof, and (b) a lipid entity linked to the chemerin or a fragment or analog thereof. Without being bound by theory, the pharmacological properties of the chemerin or a fragment or analog thereof can be modulated by the selection of the lipid entity. In some embodiments, the compositions of the present disclosure can function as agonists of CMKLR1.

[0068] In some embodiments, the lipidated chemerin composition comprising a chemerin fragment consists of the sequence YFPGQFAFS (SEQ ID NO: 2). * -FLPS * -QFA * -Tic-S (SEQ ID NO: 3).

[0069] The chemerin analog of SEQ ID NO:3 has been found to be resistant to proteolysis. See Shimamura et al., "Identification of a stable chemerin analog with potent activity toward ChemR23," Peptides 30, 2009, 1529-1538, the contents of which are incorporated by reference.

[0070] The ophthalmic formulations of the present disclosure may be pharmaceutical compositions that may further comprise a pharmaceutically acceptable carrier. Techniques for formulating the compositions of the present disclosure are described in Remington: The Science and Practice of Pharmacy, 1999. th edition, Mack Publishing Co., Easton, PA (1995). Pharmaceutical compositions can be formulated for a variety of routes of administration.

[0071] Formulations for topical administration may further comprise one or more additional ingredients.

[0072] Treatment method The present disclosure also provides a method of treating an inflammatory condition in a subject in need thereof, comprising topically administering to the eye of the subject a therapeutically effective amount of a formulation disclosed herein.

[0073] The present disclosure also provides a method of treating pain in a subject in need thereof, comprising topically administering to the eye of the subject a therapeutically effective amount of a formulation disclosed herein.

[0074] The formulations of the present disclosure can be used to treat a variety of inflammatory conditions, including, but not limited to, ocular inflammation, dry eye disease (DED), uveitis, allergic conjunctivitis, or retinal inflammatory diseases and ocular neuropathic pain.

[0075] In some embodiments, the inflammatory condition is ocular inflammation. In some embodiments, the ocular inflammation is uveitis. Uveitis is a widespread inflammatory disease of the eye, specifically the uvea. The eye has three basic layers: the sclera and cornea on the outside, the retina on the inside, and the uvea between them. The uvea is primarily composed of blood vessels and connective tissue and contains pigment cells. The various parts of the uvea are the iris in the front, the ciliary body in the middle, and the choroid, which is located behind these and surrounds most of the eye. Uveitis can affect parts of the eye other than the uvea, such as the retina, vitreous, or optic nerve. The type of uveitis is based on which part of the eye is affected. For example, anterior uveitis is inflammation of the front of the eye, called iritis or iridocyclitis; intermediate uveitis is inflammation of the middle part of the eye, or pars planitis or vitritis; posterior uveitis is inflammation of the back of the eye, such as choroiditis, retinal vasculitis, retinitis, neuroretinitis, retinochoroiditis, or chorioretinitis.

[0076] Symptoms of uveitis generally include redness, blurred vision, pain, light sensitivity, and floaters and flashes of light.

[0077] Ocular inflammation can be diagnosed by reviewing the medical history, slit lamp examination, blood tests, or any combination thereof.

[0078] Current therapies for treating ocular inflammation include topically administered anti-cytokine or anti-inflammatory agents, hi some embodiments, the formulations of the present disclosure can be administered in combination with anti-cytokine or anti-inflammatory agents for treating ocular inflammation.

[0079] Anti-cytokine or anti-inflammatory agents include, but are not limited to, NF-kappa B inhibitors, glucocorticoids such as corticosteroids, fluocinolonone; nonsteroidal anti-inflammatory drugs (NSAIDs) such as sulindac and tepoxalin; antioxidants such as dithiocarbamates; and other compounds such as sulfasalazine [2-hydroxy-5-[-4-[C2-pyridinylamino)sulfonyl]azo]benzoic acid], clonidine, and autologous blood-derived products such as Orthokine.

[0080] In some embodiments, the inflammatory condition is DED. DED is primarily caused by breakdown of the tear film in the anterior part of the eye, resulting in dehydration of the exposed outer surface. People with DED may experience stinging, gritty, itchy, or burning sensation in the eye; a foreign body sensation in the eye; watery eyes; and blurred vision. The definition and classification of DED can be found in "The Definition and Classification of Dry Eye Disease: Report of the Definition and Classification Subcommittee of the International Dry Eye Workshop (2007)", Ocular Surface 2007, Vol. 5, 75-92, the contents of which are incorporated herein by reference.

[0081] DED can be diagnosed with a comprehensive eye examination. The examination, which focuses on assessing the quantity and quality of tears produced by the eye, may include: (a) a patient history to determine the patient's symptoms and note any general health issues, medications, or environmental factors that may be contributing to the dry eye problem; (b) an external examination of the eye, including eyelid structure and blink dynamics; (c) an evaluation of the eyelids and cornea using bright light and magnification; and (d) measurement of tear quantity and quality for any abnormalities. A special dye may be placed in the eye to better observe tear flow and highlight any changes to the outer surface of the eye caused by insufficient tears.

[0082] Without being bound by theory, it is theoretically believed that ocular inflammation as a result of pro-inflammatory cytokines and growth factors plays a major role in the underlying cause of DED. Therefore, topically administered anti-cytokine or anti-inflammatory agents are often used to treat DED. In some embodiments, the pharmaceutical composition of the present disclosure, or a composition comprising chemerin or a fragment or analog thereof, can be administered in combination with an anti-cytokine or anti-inflammatory agent to treat DED.

[0083] In some embodiments, the inflammatory condition is ocular neuropathic pain. Ocular neuropathic pain can be caused by inflammation. Therefore, it can be treated with the pharmaceutical compositions of the present disclosure, optionally in combination with an anti-cytokine or anti-inflammatory agent. Typical symptoms of neuropathic pain include dysesthesia (spontaneous or evoked burning pain, often accompanied by lancinating pain), but the pain can also be deep and aching. Other sensations, such as hyperesthesia, hyperalgesia, allodynia (pain with normal stimuli), and hyperalgesia (a particularly unpleasant, exaggerated pain response), can also occur.

[0084] Methods for diagnosing ocular inflammation can be found in Teoh and Dick, "Diagnostic techniques for inflammatory eye disease: past, present and future: a review," BMC Ophthalmology 2013, 13:41, the contents of which are incorporated herein by reference.

[0085] With respect to combination therapy involving a first therapeutic agent (e.g., a formulation of the present disclosure comprising chemerin or a fragment or analog thereof) and a second therapeutic agent (e.g., an anti-cytokine or anti-inflammatory agent), the first therapeutic agent can be administered simultaneously with the second therapeutic agent; the first therapeutic agent can be administered before the second therapeutic agent; or the first therapeutic agent can be administered after the second therapeutic agent. Administration of the first and second therapeutic agents can be performed within minutes or hours, e.g., about 1, 2, 3, 4, 5, or 6 hours.

[0086] The therapeutically effective amount of a composition according to the present disclosure can vary within wide limits and can be determined by methods known in the art. For example, the composition can be administered according to body weight. Such dosages are adjusted to the individual requirements of each particular case, including the specific compound administered, the route of administration, the condition being treated, and the patient being treated. In another embodiment, the agent can be administered as a fixed dosage, e.g., not adjusted according to body weight. Generally, a daily dosage of about 0.5 mg to about 1000 mg should be appropriate, although the upper limit may be exceeded depending on the indication. The dosage can be about 5 mg to about 500 mg per day, e.g., about 5 mg to about 400 mg, about 5 mg to about 300 mg, or about 5 mg to about 200 mg. The daily dosage can be administered as a single dose or in divided doses, or, in the case of parenteral administration, as a continuous infusion. The formulations of the present disclosure can be administered once a day or several times a day, e.g., twice or three times a day. In some embodiments, the formulations of the present disclosure are administered twice daily.

[0087] In some embodiments, the therapeutically effective amount of an ophthalmic solution comprising a lipidated chimerin composition provided herein is about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, or about 0.6 mL. In some embodiments, the therapeutically effective amount of an ophthalmic solution provided herein is about 0.3 mL.

[0088] Dosage regimens utilizing the formulations of the present disclosure can be selected according to a variety of factors, including the species, ethnicity, age, weight, sex, and condition of the patient; the severity of the condition to be treated, the route of administration, the patient's renal and hepatic function; and the particular composition used. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter, or arrest the progress of the condition.

[0089] A therapeutically effective amount of a composition is that amount that provides an objectively identifiable improvement noted by a clinician or other qualified observer.

[0090] In some embodiments, a therapeutically effective amount for treating ocular inflammation is an amount that reduces the degree of inflammation in a subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to placebo.

[0091] In some embodiments, a therapeutically effective amount for treating DED is an amount that increases tear production in a subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or at least 150% compared to placebo.

[0092] In some embodiments, the efficacy of the formulations provided herein is measured according to the total corneal fluorescein staining score of one or more eyes using the Ora Calibra® scale relative to the pre-treatment score.

[0093] In some embodiments, the efficacy of the formulations provided herein is measured according to the Ocular discomfort score in one or more eyes using the Ora Calibra® scale relative to the pre-treatment score.

[0094] The formulations disclosed herein can be included in a container, pack, or dispenser together with instructions for administration. The compositions described herein can be administered topically. For example, the compositions can be administered in the form of eye drops. Those skilled in the art will recognize the advantages of the storage and dispensing / administration methods of the formulations disclosed herein.

[0095] In some embodiments, the formulations provided herein may be included in a kit comprising one or more ampoules. In some embodiments, the kit may comprise 2, 3, 4, 5, 6, 7, 8, 9, or 10 ampoules. In some embodiments, the kit may comprise about 3 ampoules. In some embodiments, each ampoules contains about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, or about 0.6 mL. In some embodiments, the ampoules may contain about 0.3 mL. In some embodiments, the kit may comprise one or more pouches, each containing one or more ampoules. In some embodiments, the kit may comprise one or more pouches, with two ampoules per pouch. In some embodiments, the ampoules are single-use blow-fill-seal (BFS) ampoules. In some embodiments, the ampoules are for direct administration to the eye.

[0096] The present disclosure also provides methods of making the ophthalmic formulations described herein. In some embodiments, the method of making the ophthalmic formulations described herein includes the steps of: (a) adding water for injection at a temperature of about 30° C. to about 80%-90% bulk batch weight of the formulation; (b) dissolving monobasic sodium phosphate monohydrate and water-free sodium phosphate in water to form a buffer solution; and (c) adding a chemerin fragment consisting of the sequence YFPGQFAFS (SEQ ID NO: 2) or YFPGQFAFS (SEQ ID NO: 2). * -FLPS * -QFA * A lipidated chemerin composition comprising a chemerin analog consisting of the sequence of -Tic-S (SEQ ID NO: 3), * dissolving a lipidated chemerin composition, wherein Tc represents a D amino acid, Tc represents 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, and the chemerin fragment or chemerin analog is linked to a lipid entity via a linker; (d) dissolving sodium chloride; (e) adjusting the temperature of the formulation to about 25°C; (f) adjusting the pH to about 7.4 to about 7.6 using 2N H3PO4 or 2N NaOH, if necessary; (e) filtering the formulation through a sterile filter; and (f) aseptically filling into batch containers. In some embodiments, the method of making produces a clear ophthalmic formulation at any of the steps above, e.g., steps (b), (c), (d), and / or (f). In some embodiments, the sterile filter is a redundant 0.2 μm sterilizing-grade polyethersulfone (PES) filter.

[0097] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although other methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred materials and methods are described herein. It should be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0098] The terms "peptide," "polypeptide," and "protein" are used interchangeably herein and typically refer to molecules comprising a chain of two or more amino acids (e.g., most typically L-amino acids, but also including, e.g., D-amino acids, modified amino acids, amino acid analogs, and amino acid mimetics). Peptides can be naturally occurring, synthetically produced, or recombinantly expressed. Peptides can also comprise additional groups that modify the amino acid chain, such as functional groups added via post-translational modifications. Examples of post-translational modifications include, but are not limited to, acetylation, alkylation (including methylation), biotinylation, glutamylation, glycylation, glycosylation, isoprenylation, lipoylation, phosphopantetheination, phosphorylation, selenation, and C-terminal amidation. The term peptide also includes peptides containing modifications of the amino and / or carboxyl termini. Modifications of the terminal amino group include, but are not limited to, des-amino modifications, N-lower alkyl modifications, N-di-lower alkyl modifications, and N-acyl modifications. Modifications of the terminal carboxy group include, but are not limited to, amide modification, lower alkyl amide modification, dialkyl amide modification, and lower alkyl ester modification (e.g., where lower alkyl is a C1-C4 alkyl). The term peptide also includes modifications of amino acids between the amino and carboxy termini, including, for example, but not limited to, the modifications described above. The term peptide can also include peptides modified to include one or more detectable labels.

[0099] As used herein, the phrase "amino acid residue" refers to an amino acid incorporated into a peptide by an amide bond or an amide bond mimetic.

[0100] The terminal amino acid at one end of a peptide chain typically has a free amino group (i.e., the amino-terminus or N-terminus). The terminal amino acid at the other end of the chain typically has a free carboxyl group (i.e., the carboxy-terminus or C-terminus). Typically, the amino acids comprising a peptide are numbered in order starting from the amino-terminus of the peptide and increasing toward the carboxy-terminus.

[0101] As used herein, the term "analog" refers to a mutant or variant polypeptide that has one or more amino acid modifications compared to the wild-type.

[0102] As used herein, an "amino acid modification" refers to a change in the amino acid sequence of a given amino acid sequence. Exemplary modifications include amino acid substitution, insertion, and / or deletion. For example, an "amino acid modification" at a particular position in a chemerin or fragment thereof refers to the substitution or deletion of a particular residue, or the insertion of at least one amino acid residue adjacent to the particular residue. An insertion "adjacent to" a particular residue means an insertion within one to two residues of the particular residue. The insertion may be N-terminal or C-terminal to the particular residue.

[0103] An "amino acid substitution" refers to the replacement of at least one existing amino acid residue in a predetermined amino acid sequence with another, different "replacement" amino acid residue. The replacement residue may be a "naturally occurring amino acid residue" (i.e., encoded by the genetic code) and may be selected from the group consisting of alanine (Ala); arginine (Arg); asparagine (Asn); aspartic acid (Asp); cysteine (Cys); glutamine (Gln); glutamic acid (Glu); glycine (Gly); histidine (His); isoleucine (Leu); leucine (Leu); lysine (Lys); methionine (Met); phenylalanine (Phe); proline (Pro); serine (Ser); threonine (Thr); tryptophan (Trp); tyrosine (Tyr); and valine (Val). Substitution with one or more non-naturally occurring amino acid residues is also encompassed within the definition of amino acid substitution herein. A "non-naturally occurring amino acid residue" refers to a residue other than the naturally occurring amino acid residues listed above that is capable of being covalently bonded to an adjacent amino acid residue in a polypeptide chain. Examples of non-naturally occurring amino acid residues include norleucine, ornithine, norvaline, homoserine, and other amino acid residue analogs such as those described in Ellman et al. Meth. Enzym. 202:301-336 (1991). To generate such non-naturally occurring amino acid residues, the procedures of Noren et al. Science 244:182 (1989) and Ellman et al., supra, can be used. Briefly, these procedures involve chemically activating a suppressor tRNA with the non-naturally occurring amino acid residue, followed by in vitro transcription and translation of the RNA. In some embodiments, L-amino acids can also be substituted with D-amino acids.

[0104] An "amino acid insertion" refers to the incorporation of at least one amino acid into a predetermined amino acid sequence. Insertions typically consist of the insertion of one or two amino acid residues, although the present application contemplates larger "peptide insertions," e.g., of about three to about five, or even up to about ten amino acid residues. The inserted residues may be naturally occurring or non-naturally occurring, as disclosed above.

[0105] An "amino acid deletion" refers to the removal of at least one amino acid residue from a predetermined amino acid sequence.

[0106] The term "pharmaceutical composition" refers to a mixture of a compound disclosed herein with other chemical components, such as a diluent or carrier. A pharmaceutical composition facilitates administration of a compound to an organism. A pharmaceutical composition can also be obtained by reacting a compound with an inorganic or organic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0107] As used herein, the term "pharmaceutically acceptable carrier" refers to any of the standard pharmaceutical carriers, such as phosphate buffered saline, water, emulsions (e.g., oil / water emulsions or water / oil emulsions), and various types of wetting agents. The compositions may also include stabilizers, preservatives, and adjuvants.

[0108] As used herein, the terms "treat," "treating," "treatment," and the like refer to reducing or ameliorating a disorder and / or its associated symptoms. It will be understood that treating a disorder or condition does not require that the disorder or symptoms associated therewith be completely eliminated, although not eliminated. The terms "treat," "treating," or "treatment" do not include prevention.

[0109] The term "therapeutically effective amount" refers to the amount of a compound that, when administered, is sufficient to prevent the onset of, or alleviate to some extent, one or more symptoms of the disorder, disease, or condition being treated. The term "therapeutically effective amount" also refers to the amount of a compound sufficient to elicit the biological or medical response sought by a researcher, veterinarian, physician, or clinician in a cell, tissue, system, animal, or human.

[0110] As used herein, a "subject" can be any mammal, e.g., a human, non-human primate, mouse, rat, dog, cat, cow, horse, pig, sheep, goat, camel. In a preferred embodiment, the subject is a human.

[0111] As used herein, a "subject in need" is a subject with an inflammatory condition.

[0112] As used herein, the singular forms "a," "an," and "the" include plural references unless the context dictates otherwise. Thus, for example, reference to "a solvent" includes a combination of two or more such solvents, reference to "a peptide" includes one or more peptides or mixtures of peptides, reference to "a drug" includes one or more drugs, reference to "a device" includes one or more devices, etc. As used herein, unless specifically stated otherwise or clear from context, the term "or" is understood to be inclusive and encompasses both "or" and "and."

[0113] Throughout this specification the word "comprising" or variations such as "comprises" or "comprising" will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0114] As used herein, the term "about" refers to ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1% of a recited value, e.g., amount, dose, temperature, time, percentage, etc., unless otherwise indicated.

[0115] As used herein, unless specifically stated otherwise or clear from the context, the term "about," when used in conjunction with a numerical value and / or range, generally refers to a numerical value and / or range that is close to the recited numerical value and / or range. In some cases, the term "about" can mean within ±10% of the recited value. For example, in some cases, "about 100 units" can mean within ±10% of 100 (e.g., 90 to 110). [Example]

[0116] The studies described herein provide for the development of an optimal formulation of the lipidated Chemerin product of the present disclosure (hereinafter referred to as OK-101) that exhibits long-term stability and shelf life.

[0117] Example 1: Determination of the solubility and pH stability of OK-101 in sodium phosphate buffer. Study Design and Objective: The solubility and pH stability of OK-101 at a target concentration of 0.05% w / v was tested in either 50 mM or 100 mM sodium phosphate buffer. For each concentration of sodium phosphate buffer, solubility and pH stability were determined in three separate buffer batches, each with a different starting pH.

[0118] Results and Observations: Results showed that concentrations >30 mg / mL of OK-101 were observed in three batches of 100 mM sodium phosphate buffer, with a slight decrease in pH due to the higher concentration (Figure 1A). Concentrations >25 mg / mL of OK-101 were observed in three batches of 50 mM sodium phosphate buffer, with a slight decrease in pH due to the higher concentration compared to 100 mM sodium phosphate buffer (Figure 1B).

[0119] Conclusion: The results showed that a sodium phosphate buffer concentration range of 50 mM to 100 mM was suitable for developing a stable formulation of OK-101.

[0120] Example 2: Determining the effect of osmogen addition on the stability and solubility of OK-101 formulations. Study Design and Objectives: The study described herein examined the effect of adding different osmogens (sodium chloride (NaCl) and mannitol) to meet the osmolality, pH, and solubility criteria for producing a stable formulation of OK-101. Any necessary adjustments were made with NaOH or by increasing the buffer concentration.

[0121] Results and Observations: Linearity and precision using HPLC of OK-101 in buffer against known validated standards (Figures 2A-2C). Various 0.2 micron membrane filters tested for suitability using one of the standards for the HPLC assay are shown in Figure 2D. The appearance and purity criteria of the OK-101 product used to develop the formulations of the present disclosure are shown in Table 1 below. [Table 1]

[0122] The informal stability of OK-101 in solutions containing preferred osmogens was further determined based on the conditions and schedule shown in Figure 3. The formulations were scaled up to 200 mL and filtered through 0.22 mM PES filters, and assay results were compared. No issues were observed between the unfiltered and filtered solutions. Stability was evaluated using glass vials as one of the container closures. As shown in Figure 4B, two formulations of 0.05% OK-101 w / v in 100 mM sodium phosphate buffer were tested with two different osmogens, A0018302-1 (a formulation containing 0.3% NaCl) and A0018302-2 (a formulation containing 1.4% mannitol). Both formulations were clear and colorless, as shown in Figure 4A. The formulations were maintained at either the following incubation conditions: 25°C and 60% relative humidity (25°C / 60% RH) or 40°C and 75% relative humidity (40°C / 75% RH) for periods of 1 month, 2 months, 3 months, or 6 months (Figure 4B and Figures 5A-5B). As shown in Figure 4B, a variation of approximately 0.4% was observed in the activity of the OK-101 product in the formulation containing 0.3% NaCl. Additionally, no significant changes in pH were observed in the formulation containing 0.3% NaCl under any of the above incubation conditions. Observations regarding changes in pH and activity were similar in the formulation containing 1.4% mannitol. For both the 0.3% NaCl and 1.4% mannitol formulations, minimal variation in relative retention time to the standard was observed over a 1-month period under both incubation conditions (Figure 4B), further demonstrating the stability of OK-101 in the formulations. Similar observations were made regarding % assay purity and relative changes in pH for the 1.4% mannitol formulation.

[0123] Further analysis of both the 0.3% NaCl and 1.4% mannitol formulations containing 0.05% OK-101 w / v in 100 mM sodium phosphate buffer was performed at 40 °C / 75% RH or 25 °C / 60% RH, with samples maintained at 5 °C for one month prior to analysis. The results, shown in Figures 4C-4E, indicated a 1%-1.9% change in % assay purity depending on the incubation conditions for the 0.3% NaCl formulation. Furthermore, there was minimal change in the RRT of OK-101 in 0.3% NaCl containing 0.05% OK-101 w / v in 100 mM sodium phosphate buffer when stored at 40 °C / 75% RH or 25 °C / 60% RH.

[0124] Conclusion: The results disclosed herein demonstrated that formulations containing the concentrations of NaCl disclosed herein are stable over extended periods of storage under a variety of storage conditions with respect to changes in pH, drug stability, and purity.

[0125] Example 3: Effect of different osmogens on the variation of formulation osmolality. Study Design and Objective: The study described herein further examined the effect of osmogen type and concentration (i.e., 0.3% NaCl or 1.4% mannitol) on the stability and purity of 0.05% OK-101 in a 100 mM sodium phosphate buffer-containing formulation over storage periods of 2, 3, and 6 months under storage conditions of 40°C / 75% RH or 25°C / 60% RH.

[0126] Results and Observations: The results described herein showed the following changes: a) The pH of the 0.3% NaCl formulation changed by 0% and 1.3% over the 2- and 3-month storage periods under both storage conditions. Osmolality changes of approximately 1.25% and 0.3% were observed at 25°C / 60% RH and 40°C / 75% RH storage conditions at 2 and 3 months, respectively, and 1.8% and 1.2% at 2 and 3 months, respectively (Figure 5A). Osmolality changes of approximately 1.8% were observed at 25°C / 60% RH and 8% at 40°C / 75% RH storage conditions over 6 months for the 0.3% NaCl formulation (Figure 5B). The formulations were observed to be clear and stable, as evidenced by the vial photographs in Figure 5C. The purity of OK-101 in the formulations tested remained >90% from 2 to 6 months of storage under both storage conditions, with purity ranging from 95% to 98%.

[0127] Based on the conditions and schedule shown in Figure 6A, the study was further expanded to determine the stability of OK-101 formulation A0019492-1 (0.1% OK-101 / 0.3% NaCl formulation), which contains 0.1% OK-101 product in 100 mM sodium phosphate buffer containing 0.3% NaCl, or OK-101 formulation A0019492-2 (0.1% OK-101 / 1.4% mannitol formulation), which contains 0.1% OK-101 product in 100 mM sodium phosphate buffer containing 1.4% mannitol, stored under storage conditions of 25°C / 60% RH or 40°C / 75% RH. The results described herein showed that the 0.1% OK-101 / 0.3% NaCl formulation remained clear and colorless after 1 and 2 months of storage under both the 25°C / 60% RH and 40°C / 75% RH storage conditions. The pH change of the 0.1% OK-101 / 0.3% NaCl formulation was observed to be less than 1.5% at 1 and 2 months of storage under both storage conditions. The osmolality change was observed to be approximately 0% and 1.2% at 1 and 2 months, respectively, under the 40°C / 75% RH storage condition, and approximately 0.3% at both 1 and 2 months under the 25°C / 60% RH storage condition. The purity of the 0.1% OK-101 / 0.3% NaCl formulation was observed to be >95% at all time points under both storage conditions.

[0128] Conclusion: The results disclosed herein demonstrated that formulations containing the concentrations of NaCl disclosed herein are stable over extended periods of storage under a variety of storage conditions with respect to changes in formulation osmolality.

[0129] The results of the studies described herein conclude that the lipidated chemerin compositions of the present disclosure, when formulated in the disclosed concentration ranges of NaCl and phosphate buffered saline (sodium phosphate buffer) at the disclosed pH and osmolality ranges, remain stable in terms of pH and osmolality fluctuations, appearance, and purity over a period of 1 to 6 months of storage under standard storage conditions.

[0130] Example 4: Clinical trial protocol to evaluate the efficacy and safety of an exemplary OK-101 ophthalmic solution compared to placebo in subjects with dry eye disease. The purpose of this clinical trial is to compare the safety and efficacy of exemplary OK-101 ophthalmic solution (hereinafter "OK-101 ophthalmic solution") with placebo for the treatment of the signs and symptoms of dry eye.

[0131] The clinical hypothesis for this study is that 0.1% OK-101 ophthalmic solution twice daily (BID) and 0.05% OK-101 ophthalmic solution BID would be superior to OK-101 placebo ophthalmic solution (vehicle) on the hierarchical primary endpoint of dry eye signs and symptoms as follows: • Signs: Total corneal fluorescein staining score of study eyes using the Ora Calibra® grading scale, measured by the mean change from baseline (Visit 2, Pre-Controlled Adverse Environment [CAE®]) to Visit 6. Symptoms: Ocular discomfort scores in both eyes using the Ora Calibra® grading scale, measured by the mean change from baseline (Visit 2, Pre-CAE®) to Visit 6. Clinical Trial Overview [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 3]

[0132] Approximately 240 subjects will be randomly assigned (1:1:1) to one of three groups to receive either OK-101 or placebo solution as topical eye drops administered bilaterally for 12 weeks. Subjects, sponsor, contract research organization (CRO), and site personnel will be masked to treatment assignment.

[0133] During a 14-day run-in period prior to randomization, all subjects receive OK-101 placebo eye drops (vehicle) BID in each eye.

[0134] During the screening period, two 90-minute exposures to CAE® are administered to confirm eligibility to participate in the study at Visit 1 (Day -14±1) and Visit 2 (Day 1). Subjects who qualify after the initial screening visit enter a run-in phase and self-administer vehicle BID for approximately 14 days. Those who qualify at Visit 2 (Day 1) are randomized to receive study medication in a double-blind fashion for 12 weeks. Subjects self-administer eye drops BID and complete diary assessments as directed.

[0135] CAE® exposures are performed at each visit with a Pre-CAE® assessment, an assessment during CAE®, and a Post-CAE® assessment of ocular signs and symptoms.

[0136] Study medication will be discontinued at Visit 6. Subjects will be withdrawn from the study at this visit.

[0137] The test design chart is shown below: [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5] [Table 4-6]

[0138] Efficacy endpoints For efficacy endpoints, the unit of analysis is the study eye or the "worst eye," as defined by:

[0139] Worst eye: An eye is eligible for analysis if it meets all of the inclusion criteria. At least one eye (right or left) must meet all criteria. If both eyes are eligible for analysis, the eye with the worst total corneal fluorescein staining score (Ora Calibra® scale) at pre-CAE® baseline will be selected as the study eye. If both eyes have the same total corneal fluorescein staining score at baseline, the right eye (OD) will be selected as the study eye.

[0140] safety scale Safety measures evaluated include visual acuity, slit lamp evaluation, adverse event referral, intraocular pressure, and dilated fundus examination.

[0141] Test treatment OK-101 ophthalmic solution is formulated as a sterile solution with a pH of 6.5 for topical ocular administration and is intended for clinical use. The study drug is supplied in blow-fill-seal ampoules that allow for direct product administration to the eye. Each ampule has a nominal volume of 0.3 mL.

[0142] The excipients used to manufacture OK-101 ophthalmic solution are standard excipients for use in ophthalmic solutions according to their respective United States Pharmacopoeia (USP) monographs / European Pharmacopoeia (EP) monographs.

[0143] The OK-101 ophthalmic solution placebo contains all the same excipients used in the active formulation, but without the peptide.

[0144] Description of route of administration, dosage, dosing regimen, and duration of treatment The dose and administration regimen were selected based on positive efficacy results from proof-of-concept non-clinical studies. The proposed treatment duration is 12 weeks.

[0145] Instructions for Use and Administration Subjects will receive OK-101 placebo eye drops (vehicle) at Visit 1 and will be assigned a study drug kit at Visits 2, 3, 4, and 5.

[0146] Randomized subjects will receive study medication BID in each eye. At Visit 2, subjects will self-administer one dose of study medication in the clinic.

[0147] Labeling / Packaging The test product (IP) is packaged and labeled in a clinical kit. The primary packaging for OK-101 ophthalmic solution is blow-fill-seal ampoules with a fill volume of 0.3 mL. The secondary packaging is foil pouches with three ampoules in each pouch.

[0148] Screening implementation period During the lead-in screening period, twelve 3-ampule pouches will be packaged in a two-week clinical kit. Each subject will receive one kit.

[0149] Treatment duration BID administration: During the treatment period, twelve 3-ampule pouches will be packaged in a 2-week clinical kit. Each subject will receive a total of six kits, one kit per visit at Visit 2 (Day 1) and Visit 3 (Day 15), and two kits per visit at Visit 4 (Visit 29) and Visit 5 (Day 57).

[0150] treatment group Subjects will be stratified by the following signs and symptoms:

[0151] Total corneal fluorescein staining score (Ora Calibra® scale) of the study eye at Visit 2, pre-CAE (<=5; >5).

[0152] Ocular discomfort score using the Ora Calibra® scale at Visit 2, pre-CAE (<=2; >2).

[0153] Exam period Participation for each individual subject involves six visits over approximately 99 days (14 days prescreening, 85 days treatment).

[0154] Example 5: Description of an exemplary manufacturing process and process control for OK-101 ophthalmic solution. As described in Table 2, an exemplary OK-101 ophthalmic solution (hereinafter "OK-101 ophthalmic solution") is aseptically manufactured by sterile filtration using redundant 0.2 μm sterilizing-grade polyethersulfone (PES) filters. The pH of this ophthalmic solution is 7.4-7.6, which is suitable for topical ocular administration.

[0155] OK-101 ophthalmic solution is filled / primarily packaged in single-use blow-fill-seal (BFS) ampoules that allow the product to be administered directly to the eye. Each ampule has a nominal volume of 0.3 mL. Secondary packaging is in foil pouches with two ampoules in each pouch. Development batch finished formulations were stored at 25±2°C / 60±5% relative humidity (RH). Clinical batch finished formulations will be stored at 2-8°C based on available stability data.

[0156] The manufacturing process and controls for OK-101 ophthalmic solution are performed in accordance with current good manufacturing practice. The manufacture of each active drug concentrate is similar except for the amount of drug substance added. The manufacture of the placebo drug is similar to the manufacture of both active drug concentrates except that the drug substance is omitted. [Table 5] AQL = Acceptable Quality Limit; HCl = Hydrochloric acid; NaOH = Sodium hydroxide; qs = qs or qsified; WFI = Water for injection.

[0157] Throughout this specification and the claims that follow, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" are understood to mean the inclusion of a stated integer, step, group of integers or group of steps, but not the exclusion of any other integer, group of integers or group of steps.

[0158] All patents, patent applications and references mentioned throughout the present specification are hereby incorporated by reference in their entirety.

[0159] The present invention includes all combinations of the preferred and more preferred groups, and suitable and more suitable groups, and embodiments of groups listed above.

Claims

1. An ophthalmic preparation comprising (a)about 0.1% to about 0.5% w / v of NaCl, and (b)about 25 mM to about 100 mM of phosphate buffered saline, and (c)A camellin fragment consisting of the sequence of Y-F-P-G-Q-F-A-F-S (SEQ ID NO: 2) or Y ＊ -F-L-P-S ＊ -Q-F-A ＊ A lipidated camellin composition containing a camellin analog consisting of the sequence of -Tic-S (SEQ ID NO: 3), from about 0.05% to about 0.1% w / v, ＊ wherein D represents a D-amino acid, Tic represents 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, and the camellin fragment or the camellin analog is linked to a lipid entity via a linker, a lipidated camellin composition having a pH of about 6.5 to about 8.5 and a weight osmolarity of about 200 to about 450 mOsm / Kg, An ophthalmic preparation.

2. The preparation according to claim 1, wherein the pH of the preparation changes by 1.5% or less over a period of 1 to 6 months when stored at 25°C to 40°C and a relative humidity of 60% to 75%.

3. The preparation according to claim 1 or 2, comprising NaCl at a concentration of about 0.1% to about 0.3% w / v.

4. The preparation according to claim 3, comprising NaCl at a concentration of about 0.3% w / v.

5. The preparation according to any one of claims 1 to 4, comprising phosphate buffered saline at a concentration of about 25 mM to about 50 mM.

6. The preparation according to claim 5, comprising phosphate buffered saline at a concentration of about 50 mM.

7. The preparation according to any one of claims 1 to 6, comprising the lipidated cemerin composition at a concentration of about 0.05% w / v.

8. The preparation according to any one of claims 1 to 6, comprising the lipidated cemerin composition at a concentration of about 0.1% w / v.

9. The preparation according to any one of claims 1 to 8, having a pH of about 7.4 to about 7.

6.

10. The preparation according to any one of claims 1 to 9, having a pH of about 7.2 to about 7.

4.

11. The preparation according to any one of claims 1 to 10, having a weight osmolarity of about 313 to about 322 mOsm / kg.

12. The preparation according to claim 11, having a weight osmolarity of about 314 to about 319 mOsm / kg.

13. The preparation according to any one of claims 1 to 12, wherein the pH of the preparation changes by 1% to 1.5% over a period of 6 months when stored at 25°C and a relative humidity of 60%.

14. The preparation according to any one of claims 1 to 13, wherein the pH of the preparation changes by 1% to 1.5% over a period of 6 months when stored at 40°C and a relative humidity of 75%.

15. The preparation according to any one of claims 1 to 12, wherein the pH of the preparation does not change over a period of 6 months when stored at 25°C and a relative humidity of 60%.

16. The preparation according to any one of claims 1 to 12, wherein the pH of the preparation does not change over a period of 6 months when stored at 40°C and a relative humidity of 75%.

17. ​ The weight osmolality of the preparation changes by 10% or less over a period of 1 to 6 months when stored at 25°C to 40°C and a relative humidity of 60% to 75%, the preparation according to any one of claims 1 to 12.

18. The weight osmolality of the preparation changes by 0.25% to 2% over a period of 6 months when stored at 25°C and a relative humidity of 60%, the preparation according to any one of claims 1 to 13 and 15.

19. The weight osmolality of the preparation changes by 1% to 8% over a period of 6 months when stored at 40°C and a relative humidity of 75%, the preparation according to any one of claims 1 to 12, 14 and 16 to 17.

20. The weight osmolality of the preparation does not change over a period of 6 months when stored at 25°C and a relative humidity of 60%, the preparation according to any one of claims 1 to 13 and 15.

21. The weight osmolality of the preparation does not change over a period of 6 months when stored at 40°C and a relative humidity of 75%, the preparation according to any one of claims 1 to 12, 14 and 16 to 17.

22. The purity of the lipidated cemerin composition used for preparing the preparation, determined by HPLC, is ≧94.6%, and the peptide content is ≧95.9, the preparation according to any one of claims 1 to 21.

23. The purity of the lipidated cemerin composition used for preparing the preparation is 95% to 98%, the preparation according to claim 22.

24. Tic is 【Chemical 1】 representing, the preparation according to any one of claims 1 to 23.

25. The linker is selected from the group consisting of the following, the preparation according to any one of claims 1 to 24. 【Chemical Formula 2】

26. The linker contains polyethylene glycol, GG, KGG, or a combination thereof, the preparation according to any one of claims 1 to 24.

27. The lipid entity is linked to the N-terminus or in the vicinity thereof of the cemerin fragment or the cemerin analog, the preparation according to any one of claims 1 to 26.

28. The lipid entity is linked to the C-terminus or in the vicinity thereof of the cemerin fragment or the cemerin analog, the preparation according to any one of claims 1 to 26.

29. The lipidated cemerin composition has the following structure: 【Chemical Formula 3】 having, the conjugate according to claim 1.

30. The formulation according to any one of claims 1 to 29, formulated for topical administration as an eye drop.

31. A method of treating an inflammatory condition in a subject in need thereof, the method comprising topically administering to the eye of the subject a therapeutically effective amount of the formulation according to any one of claims 1 to 30.

32. The method according to claim 30, wherein the inflammatory condition is dry eye disease, uveitis, allergic conjunctivitis, or retinal inflammatory disease.

33. A method of treating pain in a subject in need thereof, the method comprising topically administering to the eye of the subject a therapeutically effective amount of the formulation according to any one of claims 1 to 30.

34. The method according to any one of claims 31 to 33, wherein the formulation is administered once a day, twice a day, or three times a day.

35. The method according to any one of claims 31 to 34, wherein the subject is a human.

36. The method according to claim 35, wherein the formulation is administered twice a day.

37. The method according to any one of claims 31 to 36, wherein the effectiveness of the treatment is measured by total corneal fluorescein staining and / or relative eye discomfort score relative to the pre-treatment score.

38. A kit for administration to a subject in need thereof, the kit comprising one or more ampoules containing about 0.3 mL of the formulation according to claim 30, and instructions for use.

39. The kit according to claim 38, comprising one or more foil pouches, each pouch comprising two ampoules.

40. The kit according to claim 38 or 39, wherein the ampoule is a single-use blow-fill-seal (BFS) ampoule.

41. A method of preparing the formulation according to claim 30, comprising: (a) adding water for injection at a temperature of about 30°C to the formulation at about 80% to 90% of the bulk batch weight; (b) dissolving monobasic sodium phosphate monohydrate and sodium phosphate without water in water to form a buffer solution; (c) A lipidated kemptin composition comprising a kemptin fragment consisting of the sequence of Y-F-P-G-Q-F-A-F-S (SEQ ID NO: 2) or Y ＊ -F-L-P-S ＊ -Q-F-A ＊ -Tic-S (SEQ ID NO: 3) kemptin analog, wherein ＊ represents a D-amino acid, Tic represents 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, and the kemptin fragment or the kemptin analog is linked to a lipid entity via a linker, dissolving the lipidated kemptin composition; (d) dissolving sodium chloride; (e) adjusting the temperature of the formulation to about 25°C; (f) If necessary, adjusting the pH to about 7.4 to about 7.6 using 2N H ３ PO ４ or 2N NaOH, and (e) filtering the formulation through a sterile filter; (f) aseptically filling a batch container; and including.

42. The preparation method according to claim 41, wherein any one of steps (b), (c), (d), or (f) produces a clear formulation.

43. The production method according to claim 41 or 42, wherein the sterilization filter is a redundant 0.2 μm sterilization grade polyethersulfone (PES) filter.

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