WO2025179141A1 - Treatment of dry eye disease - Google Patents

Abstract

The disclosure provides methods of treating dry eye disease in a human in need of treatment thereof. The methods comprise instilling once weekly, into the eye of the human, a drop of an ophthalmic solution comprising between 0.03% and 0.05% w/v of Compound 1, or a pharmaceutically acceptable acid addition salt thereof.

Description

TREATMENT OF DRY EYE DISEASE

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority of U.S. Provisional Patent Application No. 63/556,763, filed February 22, 2024, the disclosure of which is incorporated by reference herein.

TECHNICAL FIELD

[0002] The disclosure is related to methods of treating dry eye disease.

BACKGROUND

[0003] Dry eye disease (DED) is a common, multifactorial ocular surface disorder characterized by a disruption in tear film homeostasis resulting in tear film fluid instability, hyperosmolarity, ocular surface damage, and inflammation. DED is highly prevalent and often is accompanied by visual disturbances, photophobia, foreign body sensation, itchiness, grittiness, burning, and conjunctival redness. DED-associated discomfort, and its impact on visual function can interfere with daily activities such as driving, reading, and using digital screens, dramatically affecting quality of life.

[0004] The first line of treatment for alleviating DED symptoms is over-the-counter artificial tears. Most FDA-approved pharmacologic topical therapies, including cyclosporin, lifitegrast, and corticosteroids, target inflammation. However, these therapies can be met with side effects, lack of effectiveness, and poor compliance. Due to the heterogeneity' of DED, available treatments are not effective for all patients, thus, an unmet need remains.

[0005] More effective methods for treating DED are needed.

SUMMARY

[0006] The disclosure is directed to methods of treating dry eye disease in a human in need of treatment thereof. The methods comprise, for example, instilling once weekly, into the eye of the human, a drop of an ophthalmic solution comprising between 0.03% and 0.05% w/v of Compound 1, or a pharmaceutically acceptable acid addition salt thereof:

Compound 1.

BRIEF DESCRIPTION OF THE DRAWINGS

[0007] The present application is further understood when read in conjunction with the appended drawings. For the purpose of illustrating the subject matter, there are shown in the drawings exemplary embodiments of the subject matter; however, the presently disclosed subject matter is not limited to the specific compositions and methods disclosed.

[0008] FIG. l is a schematic of the procedure of Example 1.

[0009] FIG. 2 is a bar graph Bar graph showing the mean (SE) change from baseline STT (Visit 3) in study eyes after three single weekly doses and 7 days of BID dosing.

[0010] FIG. 3A is a Bar graph showing the mean (SE) change from baseline STT (Visit 3) in companion eyes after three single weekly doses and 7 days of BID dosing (Visit 7) and FIG. 3B is a bar graph showing the mean change from baseline STT (Visit 3) after three single weekly doses (Visit 6).

[0011] FIG. 4A is a bar graph showing the mean (SE) change from baseline (Visit 2) for the at Visit 7 for superior, central, inferior, and total comeal fluorescein staining on the Ora Calibra™ scale (0-4 points per region with 0.5 increments permitted; maximum total score of 12) for the study eye. FIG. 4B is a bar graph showing the mean (SE) change from baseline (Visit 2) for the at Visit 7 for superior, central, inferior, and total comeal fluorescein staining on the Ora Calibra™ scale (0-4 points per region with 0.5 increments permitted; maximum total score of 12) for the companion eye. *P value from analysis of mean treatment difference.

[0012] FIG. 5A is a bar graph showing the mean (SE) change from Visit 3 at Visit 7 for superior, central, inferior, and total comeal fluorescein staining on the Ora Calibra™ scale (0-4 points per region with 0.5 increments permitted; maximum total score of 12) for the study eye. FIG. 5B is a bar graph showing the mean (SE) change from Visit 3 at Visit 7 for superior, central, inferior, and total comeal fluorescein staining on the Ora Calibra™ scale (0-4 points per region with 0.5 increments permitted; maximum total score of 12) for the companion eye. *P value from analysis of mean treatment difference. [0013] FIGs. 6A-6C are line graphs showing the change from baseline in key symptom scores. FIG. 6A is the eye dry ness score (EDS) of the VAS (scored on a scale of 0- 100). FIG. 6B is the ocular surface disease index (OSDI - score is calculated on a scale of 0 to 100). FIG. 6C is the ocular discomfort score (ODS) for the study eye (Ora Scale, 0-4). Error bars represent SE.

[0014] FIG. 7 is a bar graph showing mean (SE) change in STT at 30- and 120- minutes post IP-instillation compared to each Visit's baseline. *P value from analysis of mean treatment difference.

[0015] FIG. 8 is a bar graph showing replenishment tear dynamics measured at 35- and 125-min post IP-instillation. Error bars represent SE.

[0016] FIG. 9 is a line graph showing the mean (SE) change from baseline STT (Visit 3) after one (Visit 4) and two (Visit 5) single weekly doses. Error bars represent SE.

[0017] FIG. 10A is a bar graph showing the mean (SE) change from baseline (Visit

2) at Visit 7 for temporal, nasal, and total conjunctival Lissamine Green staining on the Ora Calibra™ scale (0-4 points per region with 0.5 increments permitted; maximum total score of 8) in the study eye. FIG. 10B is a bar graph showing the mean (SE) change from baseline (Visit 2) at Visit 7 for temporal, nasal, and total conjunctival Lissamine Green staining on the Ora Calibra™ scale (0-4 points per region with 0.5 increments permitted; maximum total score of 8) in the companion eye. *P value from analysis of mean treatment difference.

[0018] FIG. 11 A is a bar graph showing the mean (SE) change from baseline (Visit

3) at Visit 7 for temporal, nasal, and total conjunctival Lissamine Green staining on the Ora Calibra™ scale (0-4 points per region with 0.5 increments permitted; maximum total score of 8) in the study eye. FIG. 1 IB is a bar graph showing the mean (SE) change from baseline (Visit 3) at Visit 7 for temporal, nasal, and total conjunctival Lissamine Green staining on the Ora Calibra™ scale (0-4 points per region with 0.5 increments permitted; maximum total score of 8) in the eye.

[0019] FIGs. 12A-12F are line graphs showing the change from baseline in VAS symptoms. FIG 12A corresponds to blurred vision, FIG. 12B corresponds to buming/stinging. FIG. 12C corresponds to foreign body sensation, FIG. 12D corresponds to itching. FIG. 12E corresponds to pain, and FIG. 12F corresponds to photophobia. Error bars represent SE. [0020] FIG. 13 is a line graph showing the change from baseline in tear break-up time (TBUT).

[0021] FIG. 14 is a bar graph showing the comparison of tear osmolarity between Compound 1 and placebo groups at Visit 7.

[0022] FIG. 15 is a bar graph showing the comparison of meibum and viscosity between Compound 1 and placebo groups at Visit 7.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0023] The present disclosure may be understood by reference to the following detailed description which forms a part of this disclosure. The invention is not limited to the specific methods, conditions or parameters described and/or shown herein, and the terminology used herein is for the purpose of describing particular embodiments by way of example only and is not intended to be limiting of the claimed invention.

[0024] Scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art, unless otherwise defined herein.

[0025] Ranges described herein include the endpoints, as well as each value between the endpoints. For example, a range of from “1 to 5” includes the values 1 and 5, as well as all values between 1 and 5.

[0026] In the present disclosure the singular forms “a,” “an” and “the” include the plural reference, and reference to a particular numerical value includes at least that particular value, unless the context clearly indicates otherwise. Thus, for example, a reference to “a material” is a reference to at least one of such materials and equivalents thereof known to those skilled in the art, and so forth.

[0027] The terms “patient” or “subject” as used herein refer to a mammalian animal and are used interchangeably. In some embodiments, the patient or subject is a human. In other embodiments, the human is an adult (18 years of age or older).

[0028] “Pharmaceutically acceptable” means approved or approvable by a regulatory agency of the Federal or a state government or the corresponding agency in countries other than the United States, or that is listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly, in humans. [0029] As used herein, “baseline” refers to a human’s score, or average of scores, on a particular measure, prior to any administration of Compound 1 , as described herein. Baseline can be established, e.g., just prior to Compound 1 administration or up to about 7 days prior to Compound 1 administration. For example, baseline can be established about 30 minutes prior to the first instillation of Compound 1. In other aspects, baseline can be established 1 day prior to the first instillation of Compound 1. In other aspects, baseline can be established 2 days prior to the first instillation of Compound 1. In other aspects, baseline can be established 3 days prior to the first instillation of Compound 1. In other aspects, baseline can be established 4 days prior to the first instillation of Compound 1. In other aspects, baseline can be established 5 days prior to the first instillation of Compound 1. In other aspects, baseline can be established 6 days prior to the first instillation of Compound 1. In other aspects, baseline can be established 7 days prior to the first instillation of Compound 1.

[0030] According to the disclosure, methods of treating dry eye disease in a human in need of treatment thereof are provided using an ophthalmic solution comprising Compound 1, or a pharmaceutically acceptable acid addition salt thereof. In some embodiments, the ophthalmic solution comprises Compound 1 (z.e.. free base Compound 1). In other embodiments, the ophthalmic solution comprises a pharmaceutically acceptable acid addition salt of Compound 1.

[0031] The term “Compound 1” as used herein refers N2-methyl-N4-phenyl-6- (2,2,3,3-tetrafluoropropoxy)-I,3,5-triazine-2,4-diamine and has the following structure:

Compound 1.

Compound 1 is an activator of the cystic fibrosis transmembrane conductance regulator (CFTR) and has been described in, for example, WO-2017/112951. the entirety of which is incorporated by reference herein.

[0032] As used herein, a pharmaceutically acceptable acid addition salt of Compound 1 refers to an acid addition salt of Compound 1. Pharmaceutically acceptable acid addition salts are know n by those of skill in the art. Examples of such salts of the Formula I compounds are described in WO-2017/112951.

[0033] The ophthalmic solution used in the methods of the disclosure comprises between 0.03% and 0.05% w/v of Compound 1. In some embodiments, the ophthalmic solution comprises 0.03% w/v, 0.031% w/v, 0.032% w/v, 0.033% w/v, 0.034% w/v, 0.035% w/v, 0.036% w/v, 0.037% w/v, 0.038% w/v, 0.039% w/v, 0.04% w/v, 0.041% w/v, 0.042% w/v, 0.043% w/v, 0.044% w/v, 0.045% w/v, 0.046% w/v. 0.047% w/v, 0.048% w/v, 0.049% w/v of Compound 1. In other embodiments, the ophthalmic solution comprises 0.03 to 0.045% w/v, 0.03 to 0.04% w/v, 0.03 to 0.035% w/v, 0.035 to 0.05% w/v, 0.035 to 0.045% w/v, 0.035 to 0.04% w/v, 0.04 to 0.05% w/v, 0.04 to 0.045% w/v, or 0.045 to 0.05% w/v of Compound 1. In some aspects, the ophthalmic solution comprises 0.0272-0.0408 % w/v of Compound 1. In some aspects, the ophthalmic solution comprises 0.034% w/v of Compound 1. Suitable ophthalmic solutions for use in the described methods are also described in WO- 2021/113580, the entirety of which is incorporated by reference herein.

[0034] In addition to Compound 1, or a pharmaceutically acceptable acid addition salt thereof, the ophthalmic solution may contain one or more pharmaceutically acceptable excipients. In some embodiments, the ophthalmic solution comprises one or more of polyoxyl 40 stearate, PEG-400, propylene glycol, sodium chloride, sodium carboxymethylcellulose (CMC, e.g, Cekol 150), sodium thiosulfate pentahydrate, sodium EDTA dihydrate, sodium dihydrogen phosphate monohydrate, disodium phosphate anhydrous, and water (e.g., Water for Injection).

[0035] In some embodiments, the ophthalmic solution contains polyoxyl 40 stearate. In other embodiments, the ophthalmic solution contains about 3-8% w/v, for example, about 3, 4, 5, 6, 7, or 8% w/v of polyoxyl 40 stearate. In some aspects, the ophthalmic solution includes 4-6% w/v of polyoxyl 40 stearate. In some aspects, the ophthalmic solution includes 5% w/v of polyoxyl 40 stearate.

[0036] In further embodiments, the ophthalmic solution contains polyethylene glycol (PEG), for example, PEG-400. In yet other embodiments, the ophthalmic solution contains about 0.5-10% w/v of PEG, for example. PEG-400. Ophthalmic solutions used in the recited methods include, for example, 0.5. 1, 1.5, 2, 2.5. 3, 3.5, 4. 4.5. 5, 5.5, 6. 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10% w/v of PEG-400. In some aspects, the ophthalmic solution includes 1% w/v of PEG-400. In some aspects, the ophthalmic solution includes about 1-3% w/v of PEG- 400. In some aspects, the ophthalmic solution includes 0.8- 1.2% w/v of PEG-400. In some aspects, the ophthalmic solution includes 1% w/v of PEG-400.

[0037] In still further embodiments, the ophthalmic solution contains propylene glycol. In other embodiments, the ophthalmic solution contains about 0.5-10% w/v of propylene glycol, for example, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10% w/v of propylene glycol. In some aspects, the ophthalmic solution contains 1% w/v of propylene glycol. In some aspects, the ophthalmic solution contains 0.8-1.2% w/v of propylene glycol.

[0038] In further embodiments, the ophthalmic solution contains a tonicity agent suitable of ophthalmic use. In some aspects, the ophthalmic solution includes sodium chloride. In yet other embodiments, the ophthalmic solution contains 0. 1-1% w/v of a tonicity agent such as sodium chloride. In other aspects, the ophthalmic solution includes 0.1, 1.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1% w/v of sodium chloride. In some aspects, the ophthalmic solution includes 0.04-0.06 % w/v of sodium chloride. In some aspects, the ophthalmic solution includes 0.05% w/v of sodium chloride.

[0039] In still further embodiments, the ophthalmic solution contains a viscosity adjusting agent, for example, sodium CMC (e.g., Cekol 150). The viscosity adjusting agent can be present in amounts ranging from 0.05-0.5% w/v, for example, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, and 0.5% w/v. In other embodiments, the ophthalmic solution contains 0.24-0.36% w/v sodium CMC (e.g., Cekol 150). In other embodiments, the ophthalmic solution contains 0.3% w/v sodium CMC (e.g., Cekol 150).

[0040] In further embodiments, the ophthalmic solution contains an antioxidant or antioxidant system. In some aspects, the ophthalmic solution includes sodium thiosulfate pentahydrate. The sodium thiosulfate pentahydrate can be present in an amount of from about 0. 1-0.6% w/v, for example, about 0.1. 0.2, 0.3, 0.4, 0.5. or 0.6% w/v. In still other embodiments, the ophthalmic solution contains 0.16-0.24 % w/v sodium thiosulfate pentahydrate. In still other embodiments, the ophthalmic solution contains 0.2% w/v sodium thiosulfate pentahydrate. In yet further embodiments, the ophthalmic solution contains sodium EDTA dihydrate. The sodium EDTA dihydrate can be present in an amount of from about 0. 1-0.6% w/v. for example, about 0.1. 0.2, 0.3, 0.4. 0.5. or 0.6% w/v. In other embodiments, the ophthalmic solution contains 0.08-1.2% w/v sodium EDTA dihydrate. In other embodiments, the ophthalmic solution contains 0.1% w/v sodium EDTA dihydrate. [0041] In further embodiments, the ophthalmic solution includes a buffer or buffer system to achieve an acceptable pH. In some aspects, the ophthalmic solution contains sodium dihydrogen phosphate monohydrate, for example, between 0.005 and 0.05% w/v of sodium dihydrogen phosphate. In some aspects, the ophthalmic solution contains sodium dihydrogen phosphate monohydrate, for example, 0.0210-0.0314% w/v of sodium dihydrogen phosphate. In yet other embodiments, the ophthalmic solution contains 0.02% w/v (e.g.. 0.026% or 0.0262% w/v) sodium dihydrogen phosphate monohydrate. In still further embodiments, the ophthalmic solution contains disodium phosphate anhydrous, for example, between 0.005 and 0.5% w/v of disodium phosphate anhydrous. In still further embodiments, the ophthalmic solution contains 0.1% w/v (e.g., 0.11% or 0.115% w/v) of disodium phosphate anhydrous. In still further embodiments, the ophthalmic solution contains 0.092-0. 138% w/v of disodium phosphate anhydrous.

[0042] The ophthalmic solutions used in the described methods will also include water, for example, contains quantum sufficit (QS) to 100 mL of water.

[0043] In some aspects, ophthalmic solutions comprises:

[0044] The terms “dry eye disease” or “dry eye disorder” are used interchangeably and refer to a heterogeneous group of disorders. These terms can also refer to signs and symptoms and any disorders associated with dry eye disease. See, Shimazaki, Investigative Ophthalmology & Visual Science, November 2018, 59, DES7-DES12, which is incorporated by reference herein. In some embodiments, the dry eye disease/disorder is a disorder of the tear fdm due to tear deficiency or excessive evaporation, which causes damage to the interpalpebral ocular surface and is associated with symptoms of ocular discomfort. In other embodiments, dry eye disease/disorder is a multifactorial disease of the tears and ocular surface that results in symptoms of discomfort, visual disturbance, and tear film instability with potential damage to the ocular surface. In further embodiments, dry eye disease/disorder is a multifactorial disease of the ocular surface characterized by a loss of homeostasis of the tear film, and accompanied by ocular symptoms, in which tear film instability and hyperosmolarity, ocular surface inflammation and damage, and neurosensory abnormalities play etiological roles. In yet other embodiments, dry eye disease/disorder is ocular surface epithelial damage caused by qualitative or quantitative abnormalities of tears. In still further embodiments, dry eye disease/disorder is ocular is chronic disease in tears and comeal/conjunctival epithelia caused by various factors and may accompany irritating symptoms and/or visual disturbances. In other embodiments, dry eye disease/disorder is a multifactorial disease characterized by unstable tear film causing a variety of symptoms and/or visual impairment, potentially accompanied by ocular surface damage. Signs of dry eye include damage to the corneal epithelial cells, tear deficiency, excessive evaporation, increased osmolarity of the tear film, inflammation of the ocular surface, tear film instability, hyperosmolarity, ocular surface inflammation and damage, and neurosensory abnormalities.

[0045] In certain embodiments, the dry eye disease is aqueous deficient dry eye, evaporative dry eye, or mixed dry eye. In some embodiments, the dry eye disease is aqueous deficient dry eye. In other embodiments, the aqueous deficient dry eye is Sjogren syndrome dry eye or non-Sjogren dry eye. In other embodiments, the aqueous deficient dry eye is Sjogren syndrome dry eye, z.e., the human has Sjogren syndrome. In further embodiments, the dry eye disease is primary Sjogren syndrome dry eye. In further embodiments, the dry eye disease is secondary Sjogren syndrome dry eye. In yet other embodiments, the dry⁷ eye disease is a non-Sjogren syndrome dry eye. In still further embodiments, the non-Sjogren syndrome dry⁷ eye is lacrimal deficiency, lacrimal gland duct obstruction, reflex block, or drug-related. In other embodiments, the non-Sjogren syndrome dry eye is lacrimal deficiency In further embodiments, the non-Sjogren syndrome dry eye lacrimal gland duct obstruction. In still other embodiments, the non-Sjogren syndrome dry eye reflex block. In yet further embodiments, the non-Sjogren syndrome dry⁷ eye is drug-related, e.g., antihistamines, betablockers, antispasmodics, diuretics, psychotropics, or combinations thereof. In other embodiments, the dry eye disease is evaporative dry eye. In further embodiments, the evaporative dry' eye is intrinsic or extrinsic. In yet other embodiments, the evaporative dry⁷ eye is intrinsic, such as meibomian oil deficiency, disorders of lid aperture, low blink rate, or by drug action (e.g., Accutane. In still further embodiments, the evaporative drug eye is extrinsic, such as vitamin A deficiency, topical drugs preservatives, contact lens wear, or ocular surface disease, (e.g., allergy). In other embodiments, the dry eye disease is mixed dry eye.

[0046] In further embodiments, the dry eye disease in the human is associated with aging, contact lens usage, or refractive surgery. In yet other embodiments, the dry eye disease in the human is associated with aging. In still further embodiments, the dry eye disease in the human is associated with contact lens usage. In other embodiments, the dry eye disease in the human is associated with refractive surgery.

[0047] In further embodiments, the dry eye disease in the human is associated with a systemic immunologic disease., for example, diabetes, rheumatoid arthritis, scleroderma, and systemic lupus erythematosus.

[0048] In yet other embodiments, the dry eye disease is definite dry eye disease or probable dry eye disease. In still further embodiments, the dry eye disease is definite dry eye disease. In other embodiments, the dry eye disease is probable dry eye disease.

[0049] In further embodiments, the dry eye disease is characterized by one or more signs or symptoms of dry eye disease. Signs and symptoms of dry eye disease include, but are not limited to visual disturbances, changes in tears, epithelial damage, unstable tear film, ocular photophobia, ocular foreign body sensation, ocular itching, ocular grittiness, ocular burning, or conjunctival redness. In yet other embodiments, the dry eye disease is characterized by visual disturbances. In still further embodiments, the dry eye disease is characterized by changes in tears. In other embodiments, the dry eye disease is characterized by epithelial damage. In further embodiments, the dry eye disease is characterized by unstable tear film. In still other embodiments, the dry eye disease is characterized by ocular photophobia. In yet further embodiments, the dry eye disease is characterized by ocular foreign body sensation. In other embodiments, the dry eye disease is characterized by ocular itching. In further embodiments, the dry eye disease is characterized by ocular grittiness. In yet other embodiments, the dry eye disease is characterized by ocular burning. In still further embodiments, the dry eye disease is characterized by conjunctival redness.

[0050] Dry eye disease/disorder may be diagnosed or evaluated using a number of tests or techniques. In some embodiments, dry eye disease is diagnosed or evaluated using one or more of visual acuity procedures (e.g., ETDRS chart), slit lamp biomicroscopy procedures, intraocular pressure, dilated fundoscopy, Schirmer’s tear test with topical anesthesia, visual analogue scope-symptom index, self-reported ocular discomfort scales, self-reported ocular irritation scales, self-reported vision-related functioning scales, Tear Break-up Time, (TBUT), corneal and conjunctival staining, tear film osmolarity, meibomian gland imaging and expression, and subject-reported drop comfort evaluation. In other embodiments, dry eye disease is diagnosed or evaluated using visual acuity procedures (e.g., ETDRS chart). In further embodiments, dry eye disease is diagnosed or evaluated using slit lamp biomicroscopy procedures. In yet other embodiments, dry eye disease is diagnosed or evaluated using intraocular pressure. In still further embodiments, dry eye disease is diagnosed or evaluated using dilated fundoscopy. In other embodiments, dry eye disease is diagnosed or evaluated using Schirmer’s tear test with topical anesthesia. In further embodiments, dry eye disease is diagnosed or evaluated using visual analogue scopesymptom index. In still other embodiments, dry eye disease is diagnosed or evaluated using a self-reported ocular discomfort scale. In yet further embodiments, dry eye disease is diagnosed or evaluated using a self-reported ocular irritation scale. In other embodiments, dry eye disease is diagnosed or evaluated using a self-reported vision-related functioning scalea. In other embodiments, dry eye disease is diagnosed or evaluated using TBUT. In further embodiments, dry eye disease is diagnosed or evaluated using comeal and conjunctival staining. In yet other embodiments, dry eye disease is diagnosed or evaluated using tear film osmolarity. In still further embodiments, dry eye disease is diagnosed or evaluated using meibomian gland imaging and expression. In yet further embodiments, dry eye disease is diagnosed or evaluated using a subject-reported drop comfort evaluation scale. See, e.g., the tests and techniques described in Example 2.

[0051] In certain embodiments, the human has an anesthetized Schirmer’s tear test (STT) of between 5 and 10 mm/5 min in the eye at baseline. In some embodiments, the STT is 5 mm/5 min, 6 mm/5 min, 7 mm/5 min, 8 mm/5 min, 9 mm/5 min, or 10 mm/5 min in the eye at baseline. In other embodiments, the STT is 5 to 9 mm/5 min, 5 to 8 mm/5 min, 5 to 7 mm/5 min, 5 to 6 mm/5 min, 6 to 10 mm/5 min, 6 to 9 mm/5 min, 6 to 8 mm/5 min, 6 to 7 mm/5 min, 7 to 10 mm/5 min, 7 to 9 mm/5 min, 7 to 8 mm/5 min, 8 to 10 mm/5 min, 8 to 9 mm/5 min, or 9 to 10 mm/5 min in the eye at baseline. [0052] In further embodiments, the human has a calculated best-corrected visual acuity of at least about 0.7 logarithm of the minimum angle of resolution (logMAR) in the eye at baseline.

[0053] In still further embodiments, the human has a corneal fluorescein staining in at least one region of the eye, at baseline.

[0054] According to the disclosure, the methods comprise instilling once weekly, into the eye of the human, a drop of an ophthalmic solution comprising Compound 1, or a pharmaceutically acceptable acid addition salt thereof. As used herein, a “drop” of the ophthalmic solution is about 40 pL. In certain embodiments, the drop of the ophthalmic solution is instilled once weekly, unilaterally (i.e., one eye), into the eye of the human. In further embodiments, the drop is instilled into the right eye. In other embodiments, the drop is instilled into the left eye.

[0055] In yet further embodiments, the drop of the ophthalmic solution is instilled once weekly, bilaterally, into the eyes (i.e., both) of the human.

[0056] The term “once weekly” as used herein refers to a time that is once in a period of six to eight calendar days, preferably seven calendar days. By way of example, the drop is instilled the same day of every week. The disclosure accounts for variations to instillation due to minor errors in human compliance.

[0057] The method is performed for a treatment period of at least one week. In some embodiments, the method is performed for a treatment period of at least two weeks. In other embodiments, the method is performed for a treatment period of at least three weeks. In other embodiments, the method is performed for a treatment period of at least 4, 5, 6, 7, 8, 9, or 10 weeks.

[0058] In some aspects, after the once-weekly treatment period, the drop of the ophthalmic solution is instilled twice daily, into the eye of the human. In some embodiments, the twice daily instillation is performed unilaterally, into the eye of the human. In other embodiments, the twice daily instillation is performed bilaterally into the eyes.

[0059] In certain embodiments, the disclosure provides methods of treating dry eye disease in a human in need of treatment thereof, comprising instilling twice daily, into the eye of the human, a drop of an ophthalmic solution comprising between 0.03% and 0.05% w/v of Compound 1 , or a pharmaceutically acceptable acid addition salt thereof. [0060] According to the disclosure, the treatment methods result in a number of improvements in the human. The “improvement” is related to the dry eye disease and may be determined by one skilled in the art as measured by any number of means. For example, the improvement might be evaluated using one or more of visual acuity procedures (e.g., ETDRS chart), slit lamp biomicroscopy procedures, intraocular pressure, dilated fundoscopy, Schirmer’s tear test with topical anesthesia, visual analogue scope-symptom index, selfreported ocular discomfort scales, self-reported ocular irritation scales, self-reported vision- related functioning scales, Tear Break-up Time, (TBUT), corneal and conjunctival staining, tear fdm osmolarity, meibomian gland imaging and expression, meibomian gland viscosity, and subject-reported drop comfort evaluation. In those aspects wherein an improvement is measured as compared to a control ophthalmic solution that does not include Compound 1, the improvement will be statistically significant, e.g., p=0.05, to one skilled in the art.

[0061] In some embodiments, the treatment results in an improvement in at least one sign of dry eye disease in the human. For example, signs of dry eye disease can be measured by a person skilled in the art, for example, low tear film and corneal staining. In other embodiments, the treatment results in an improvement in at least one symptom of dry eye disease in the human. For example, symptoms of dry eye disease are self-reported by the human. In further embodiments, the treatment results in an improvement in the eye of the human of one or more of eye dryness, burning, stinging, itching, foreign body sensation, blurred vision, photophobia, or pain. In yet other embodiments, the treatment results in an improvement in eye dryness. In still further embodiments, the treatment results in an improvement in eye burning. In other embodiments, the treatment results in an improvement in eye stinging. In further embodiments, the treatment results in an improvement in eye itching. In still other embodiments, the treatment results in an improvement in foreign body sensation in the eye. In yet further embodiments, the treatment results in an improvement in blurred vision. In other embodiments, the treatment results in an improvement in photophobia. In further embodiments, the treatment results in an improvement in eye pain.

[0062] In certain embodiments, the method results in an improvement in the human’s anesthetized STT, as compared to baseline.

[0063] In some embodiments, the method results in an improvement in Corneal Fluorescein Staining, as compared to baseline. [0064] In certain embodiments, the method results in an improvement in the human’s self-reported ocular discomfort scale, as compared to baseline.

[0065] In certain embodiments, the method results in an improvement in the human’s self-reported ocular irritation scale.

[0066] In certain embodiments, the method results in an improvement in the human’s self-reported vision-related functioning scale.

[0067] In some embodiments, the method results in an improvement in the human’s Eye Dryness Score (EDS) of the Visual Analogue Scale (VAS), as compared to baseline.

[0068] In some embodiments, the method results in an improvement in meibomian gland secretion viscosity, as compared to baseline.

[0069] Aspects

[0070] Aspect 1. A method of treating dry eye disease in a human in need of treatment thereof, comprising instilling once weekly, into the eye of the human, a drop of an ophthalmic solution comprising between 0.03% and 0.05% w/v of Compound 1, or a pharmaceutically acceptable acid addition salt thereof:

Compound 1.

[0071] Aspect 2. The method of Aspect 1, wherein the treatment results in an improvement in at least one sign of dry eye disease in the human.

[0072] Aspect 3. The method of any one of Aspect 1 or Aspect 2, wherein the treatment results in an improvement in at least one symptom of dry eye disease in the human.

[0073] Aspect 4. The method of any one of the preceding Aspects, wherein the treatment results in an improvement in the eye of the human of one or more of eye dryness, burning, stinging, itching, foreign body sensation, blurred vision, photophobia, or pain.

[00741 Aspect 5. The method of any one of the preceding Aspects, wherein the dry eye disease is aqueous deficient dry eye, evaporative dry eye, or mixed dry eye.

[0075] Aspect 6. The method of any one of the preceding Aspects, wherein the human has Sjogren’s syndrome. [0076] Aspect 7. The method of any one of the preceding Aspects, wherein the dry eye disease in the human is associated with aging, contact lens usage, or refractive surgery.

[0077] Aspect 8. The method of any one of the preceding Aspects, wherein the dry eye disease in the human is associated with a systemic immunologic disease.

[0078] Aspect 9. The method of any one of the preceding Aspects, wherein the dry eye disease is definite dry eye disease or probable dry eye disease.

[0079] Aspect 10. The method of any one of the preceding Aspects, wherein the human has an anesthetized Schirmer’s tear test (STT) of between 5 and 10 mm/5 min in the eye at baseline.

[0080] Aspect 11. The method of any one of the preceding Aspects, wherein the method results in an improvement in the human’s anesthetized Schirmer’s tear test (STT), as compared to baseline.

[0081] Aspect 12. The method of any one of the preceding Aspects, wherein the method results in an improvement in Corneal Fluorescein Staining, as compared to baseline.

[0082] Aspect 13. The method of any one of the preceding Aspects, wherein the method results in an improvement in the human’s self-reported ocular discomfort, as compared to baseline.

[0083] Aspect 14. The method of any one of the preceding Aspects, wherein the method results in an improvement in the human’s Eye Dryness Score (EDS) of the Visual Analogue Scale (VAS), as compared to baseline.

[0084] Aspect 15. The method of any one of the preceding Aspects, wherein the method results in an improvement in the human’s self-reported ocular irritation, as compared to baseline.

[0085] Aspect 16. The method of any one of the preceding Aspects, wherein the method results in an improvement in meibomian gland secretion viscosity, as compared to baseline.

[0086] Aspect 17. The method of any one of the preceding Aspects, wherein the human has a calculated best-corrected visual acuity of at least about 0.7 logarithm of the minimum angle of resolution (logMAR) in the eye at baseline.

[0087] Aspect 18. The method of any one of the preceding Aspects, wherein the human has a corneal fluorescein staining in at least one region of the eye, at baseline. [0088] Aspect 19. The method of any one of the preceding Aspects, wherein the dry eye disease is characterized by one or more of visual disturbances, changes in tears, epithelial damage, unstable tear film, ocular photophobia, ocular foreign body sensation, ocular itching, ocular grittiness, ocular burning, or conjunctival redness.

[0089] Aspect 20. The method of any one of the preceding Aspects, wherein the human is 18 years of age or older.

[0090] Aspect 21. The method of any one of the preceding Aspects, wherein the ophthalmic solution comprises a pharmaceutically acceptable acid addition salt of Compound 1.

[0091] Aspect 22. The method of any one of the preceding Aspects, wherein the ophthalmic solution comprises Compound 1.

[0092] Aspect 23. The method of any one of the preceding Aspects, wherein the ophthalmic solution comprises 0.034% w/v of Compound 1.

[0093] Aspect 24. The method of any one of the preceding Aspects, wherein the ophthalmic solution comprises:

[0094] Aspect 25. The method of any one of the preceding Aspects, wherein the drop of the ophthalmic solution is instilled once weekly, unilaterally, into the eye of the human.

[0095] Aspect 26. The method of any one of the preceding Aspects, wherein the drop of the ophthalmic solution is instilled once weekly, bilaterally, into the eyes of the human.

[0096] Aspect 27. The method of any one of the preceding Aspects, wherein the method is performed for a treatment period of at least one week. [0097] Aspect 28. The method of any one of the preceding Aspects, wherein the method is performed for a treatment period of at least two weeks.

[0098] Aspect 29. The method of any one of the preceding Aspects, wherein the method is performed for a treatment period of at least three weeks.

[0099] Aspect 30. The method of any one of Aspects 26-28, wherein after the treatment period, the drop of the ophthalmic solution is instilled twice daily, into the eye of the human.

[00100] Aspect 31. The method of Aspect 29, wherein the twice daily instillation is performed unilaterally, into the eye of the human.

[00101] Aspect 32. The method of Aspect 29, wherein the twice daily instillation is performed bilaterally into the eyes of the human.

[00102] Aspect 33. A method of treating dry eye disease in a human in need of treatment thereof, comprising instilling twice daily, into the eye of the human, a drop of an ophthalmic solution comprising between 0.03% and 0.05% w/v of Compound 1, or a pharmaceutically acceptable acid addition salt thereof:

Compound 1.

[00103] The following Examples are provided to illustrate some of the concepts described within this disclosure.

EXAMPLES

[00104] Example 1

[00105] A. Objectives

[00106] (i) Primary Objectives

[00107] To evaluate the efficacy of Compound 1 0.034% compared to vehicle for the treatment of the signs of dry eye as measured by the improvement in STT (pre-IP instillation) at Visit 6 or 7 compared to baseline (Visit 3).

[00108] (ii) Secondary Objectives [00109] To evaluate the effects of Compound 1 0.034% compared to vehicle on additional signs and symptoms of dry eye as measured by the following:

• Improvement in Corneal Fluorescein Staining evaluated at Visit 3, 4, 5, 6 or 7.

• Improvement in self-reported ocular discomfort evaluated by the Subject at Visit 4, 5, 6 or 7.

• Improvement in self-reported ocular irritation, evaluated by the Subject at Visit 4, 5, 6 or 7.

• Improvement in self-reported vision-related functioning evaluated by the Subject at Visit 4, 5, 6 or 7.

• Improvement in EDS of the VAS evaluated by the Subject at Visit 4, 5, 6 or 7.

[00110] B. Investigational plan

[00111] Overall Study Design

[00112] This is a single-center, double-masked, randomized, placebo-controlled, Phase II dry eye study evaluating the efficacy of Compound 1 0.034% (topical ocular, bilaterally) compared to vehicle. See, FIG. 1. At Visit 3, subjects will be randomly assigned in a 1: 1 allocation ratio to one of the following treatment arms:

• Compound 1 0.034% Ophthalmic Solution

• Vehicle of Compound 1 Ophthalmic Solution

[00113] This study will consist of three phases: a Screening Phase, a weekly Treatment Phase, and a BID Treatment Phase. The Screening Phase will consist of two visits (Visits 1 and 2) over a one to two-week period. Additional screening will occur at the beginning of Visit 3 prior to subject randomization and treatment. The weekly Treatment Phase will occur over a three-week period where dosing will occur at Visits 3, 4, 5, and 6. The BID Treatment Phase will occur over a one-week period and begin at Visit 6 and continue until Visit 7. Each type of assessment across the study will occur at the same chronological time as much as possible and within a three-hour time window.

[00114] (i) Screening Phase

[00115] (a) Visit 1 (Day -14 to -7): Informed Consent

[00116] Medical/medication history, demographics, and inclusion /exclusion review may be performed prior to Visit 1 but must be confirmed at Visit 1 (with the exception of demographics). [00117] All demographic data, medical history, any medications, and any underlying condition(s) will be collected. Current underlying conditions, including those that began within the last 45 days, which may have been resolved before screening, must be recorded. Any medications the subject is taking, as well as those the subject may have taken but discontinued within 45 days prior to Visit 1 will be recorded. It will be confirmed if a subject needs to washout from any current medications and instruct them to follow the appropriate washout time periods. All Females of childbearing potential will be given a Urine Pregnancy Test and must have a negative Urine Pregnancy Test to continue in the study and must agree to use an adequate method of contraception for the duration of the study in order to be enrolled.

[00118] Subjects will have their Initial Best-Corrected Visual Acuity assessed utilizing and ETDRS Chart. Subjects must have a score of 0.7 logMAR or better in each eye in order to qualify. This initial visual acuity will be considered the Baseline Visual Acuity. The use of correction will be documented in the source document. If a subject uses correction at this visual acuity, then they should use the same correction throughout all subsequent visual acuity assessments.

[00119] Subjects will be instructed to evaluate their symptoms (eye dryness, burning/stinging, itching, foreign body sensation, blurred vision, photophobia, and pain) using a 100-point scale (Visual Analogue Scale). Subjects will be instructed to complete questionnaire(s) to assess ocular irritation. Questionnaires used for subject self-reporting of ocular discomfort, ocular irritation, and vision-related functioning are available in the art and include questions asking the subject to report on items such as sensitivity to light, blurred vision, eye grittiness, problems reading or driving, problems watching television, uncomfortable feelings in the eyes in low humidity or in air conditioning, and the like, on a 5- point Likert scales. Digital photographs may be taken of the entire eye.

[00120] A STT will be performed in both eyes. Fluorescein eye stain will be instilled in each eye and TBUT will be measured by to assess tear film stability. Corneal and conjunctival fluorescein staining will be assessed and scored. Lissamine green will be instilled in each eye and corneal and conjunctival staining will be assessed and scored. Slitlamp photographs of corneal and conjunctival staining (fluorescein and lissamine green) will be taken of each eye. Subjects will be given an Adverse Event Query. [00121] Subjects will have IOP measured in each eye by contact tonometry' and a dilated fundus examination will be performed to evaluate the presence or absence of clinically significant fundus abnormalities and vitreous pathology. Qualifying subjects will be scheduled to return to the office in no less than 48 hours but no more than eleven days for Visit 2. Subjects must meet the following criteria to qualify to return for Visit 2: have an anesthetized STT between 5- 10mm in at least one eye and no greater than 10mm in the remaining eye, and comeal fluorescein staining in at least one region in both eyes to qualify to return for Visit 2.

[00122] (b) Visit 2 (Day -5 to -3)

[00123] Visit 2 should occur at least two days (i.e. 48 hours) and no more than eleven days after Visit 1 has occured. Visit 2 should also occur at least three days but no more than five days before Visit 3. When subjects return to the clinic, they will have the following assessments: Adverse Event Query, Update of Medical/Medication History, Urine Pregnancy Test (for females of childbearing potential), Visual Acuity Utilizing ETDRS Chart.

[00124] Subjects will be instructed to evaluate their symptoms (eye dryness, buming/stinging, itching, foreign body sensation, blurred vision, photophobia, and pain) using the VAS, complete the self-reporting questionnaire(s) (e g., for ocular discomfort, ocular irritation, and/or vision-related functioning), sequentially.

[00125] A slit-lamp examination will be performed in both eyes to exclude subjects with disallowed ocular conditions. Digital photographs may be taken of the entire eye.

[00126] For each qualified subject, a second STT will be performed in both eyes. Fluorescein eye stain will be instilled in each eye and TBUT will be measured to assess tear film stability. Comeal and conjunctival fluorescein staining will be assessed and scored. Lissamine green will be instilled in each eye and comeal and conjunctival staining will be assessed and scored. Slit-lamp photographs of comeal and conjunctival staining (fluorescein and lissamine green) will be taken of each eye. Subjects will be given an Adverse Event Query.

[00127] Subjects must meet the following criteria to qualify to return for Visit 3: have two anesthetized STT between 5- 10mm in at least one eye and no greater than 10mm in the remaining eye at Visit 1 and 2, and comeal fluorescein staining in at least one region in both eyes at Visit 1 and 2. A review of protocol inclusion and exclusion criteria will be confirmed for each subject.

[00128] (ii) Weekly Treatment Phase

[00129] (a) Visit 3 (Day 1): Additional Screening/Enrollment/Randomization/In- office Instillation

[00130] Visit 3 should occur at least three days but no more than five days after Visit 2 and at least one week but no more than two weeks after Visit 1.

[00131] Subjects will be instructed to evaluate their symptoms (eye dryness, buming/stinging, itching, foreign body sensation, blurred vision, photophobia, and pain) using the VAS, complete the self-reporting questionnaire(s) (e.g., for ocular discomfort, ocular irritation, and/or vision-related functioning, sequentially.

[00132] A slit-lamp examination will be performed in both eyes to exclude subjects with disallowed ocular conditions. Digital photographs may be taken of the entire eye.

[00133] A baseline STT will be performed at 30 minutes (± 1 minute) before product instillation and prior to subject randomization.

[00134] To qualify for the study, subjects must have three anesthetized STTs between 5-10mm at Visits 1, 2, and 3 in the study eye and no greater than 10mm in the companion eye.

[00135] A STT will be performed at 30 minutes (± 1 minute) and 35 minutes (± 2 minutes) post IP instillation. A STT will be performed again at 120 minutes (± 1 minute) and 125 minutes (± 2 minutes) post IP instillation. The 35 and 125 min STTs will measure tear replenishment dynamics and will be performed immediately after the 30 and 120 min tests.

[00136] A slit-lamp biomicroscopy exam will be performed post-STT in both eyes. A new or worsening ocular finding will be documented as a TEAE. Fluorescein eye stain will be instilled in each eye and TBUT will be measured to assess tear film stability. Comeal and conjunctival fluorescein staining will be assessed and scored. Lissamine green will be instilled in each eye and comeal and conjunctival staining will be assessed and scored. Slitlamp photographs of comeal and conjunctival staining (fluorescein and lissamine green) will be taken of each eye.

[00137] (b) Visit 4 (Day 8 ±1) and Visit 5 (Day 15 ±1): In-office Instillation

[00138] Subjects will be instructed to evaluate their symptoms (eye dryness, buming/stinging, itching, foreign body sensation, blurred vision, photophobia, and pain) using the VAS. complete the self-reporting questionnaire(s) (e.g., for ocular discomfort, ocular irritation, and/or vision-related functioning, sequentially.

[00139] A baseline STT will be performed at 30 minutes (± 1 minute) before product instillation. A trained study technician will instill the assigned investigational product according to the directions for use 30 minutes (± 1 minute) post STT.

[00140] A STT will be performed at 30 minutes (± 1 minute) and 35 minutes (± 2 minutes) post IP instillation. A STT will be performed again at 120 minutes (± 1 minute) and 125 minutes (± 2 minutes) post IP instillation. The 35 and 125 min STTs will measure tear replenishment dynamics and will be performed immediately after the 30 and 120 min tests.

[00141] Fluorescein eye stain will be instilled in each eye and TBUT will be measured gator to assess tear fdm stability. Comeal and conjunctival fluorescein staining will be assessed and scored. Lissamine green will be instilled in each eye and corneal and conjunctival staining will be assessed and scored. Slit-lamp photographs of comeal and conjunctival staining (fluorescein and lissamine green) will be taken of each eye.

[00142] (iii) BID Treatment Phase

[00143] (a) Visit 6 (Day 22 ±1): In-office Instillation/BID Dosing Instructions

[00144] Subjects will be instructed to evaluate their symptoms (eye dryness, buming/stinging, itching, foreign body sensation, blurred vision, photophobia, and pain) using the VAS, complete the self-reporting questionnaire(s) (e.g., for ocular discomfort, ocular irritation, and/or vision-related functioning, sequentially.

[00145] A baseline STT will be performed at 30 minutes (± 1 minute) before product instillation. A trained study technician will instill the assigned investigational product according to the directions for use 30 minutes (± 1 minute) post STT.

[00146] A STT will be performed at 30 minutes (± 1 minute) and 35 minutes (± 2 minutes) post IP instillation. A STT will be performed again at 120 minutes (± 1 minute) and 125 minutes (± 2 minutes) post IP instillation. The 35 and 125 min STTs will measure tear replenishment dynamics and will be performed immediately after the 30 and 120 min tests.

[00147] Fluorescein eye stain will be instilled in each eye and TBUT will be measured to assess tear film stability. Corneal and conjunctival fluorescein staining will be assessed and scored. Lissamine green will be instilled in each eye and comeal and conjunctival staining will be assessed and scored. Slit-lamp photographs of comeal and conjunctival staining (fluorescein and lissamine green) will be taken of each eye. [00148] Subjects will be provided instructions and IP for BID dosing and scheduled to return to the office exactly one week later for Visit 7.

[00149] (b) At Home BID Dosing (Day 22 ±1 to Day 28 ±1)

[00150] Subjects will receive instructions and IP at the end of Visit 6. Subjects will instill one drop in each eye in the evening (PM) on the day of Visit 6. Subjects will continue to instill one drop in each eye twice a day, once in the morning (AM) and once in the evening (PM), for 6 more days. No product will be administered by the subject on the day of Visit 7.

[00151] (c) Visit 7 (Day 29 ±1): In-office Instillation/Exit

[00152] Subjects will be instructed to evaluate their symptoms (eye dryness, buming/stinging, itching, foreign body sensation, blurred vision, photophobia, and pain) using the VAS, complete the self-reporting questionnaire(s) (e.g., for ocular discomfort, ocular irritation, and/or vision-related functioning, sequentially.

[00153] Digital photographs may be taken of the entire eye.

[00154] A baseline STT will be performed at 30 minutes (± 1 minute) before product instillation. A trained study technician will instill the assigned investigational product according to the directions for use 30 minutes (± 1 minute) post STT.

[00155] A STT will be performed at 30 minutes (± 1 minute) and 35 minutes (± 2 minutes) post IP instillation. A STT will be performed again at 120 minutes (± 1 minute) and 125 minutes (± 2 minutes) post IP instillation. The 35 and 125 min STTs will measure tear replenishment dynamics and will be performed immediately after the 30 and 120 min tests.

[00156] Fluorescein eye stain will be instilled in each eye and TBUT will be measured to assess tear film stability. Comeal and conjunctival fluorescein staining will be assessed and scored. Lissamine green will be instilled in each eye and comeal and conjunctival staining will be assessed and scored. Slit-lamp photographs of comeal and conjunctival staining (fluorescein and lissamine green) will be taken of each eye.

[00157] A tear film osmolarity measurement will be taken of each eye.

[00158] Imaging will be performed on the meibomian glands of the lower lids using slit lamp biomicroscopy. Any abnormalities of the meibomian glands will be recorded. Expression of the middle five meibomian glands on the lower lids will be performed and the color, turbidity, and viscosity of the meibum will be recorded.

[00159] Subjects will be asked to complete an Adverse Event Query and will be exited from the study [00160] Example 2: Representative Dry Eye Disease Tests

[00161] LogMAR visual acuity (VA) will be assessed using an Early Treatment Diabetic Retinopathy Study (ETDRS) chart. The procedure used will be consistent with the recommendations provided for using the ETDRS eye chart. VA should be evaluated at the beginning of each visit in the study (i.e., prior to slit lamp examination). VA testing should be done with the habitual correction.

[00162] Slit lamp biomicroscopy procedures

[00163] This examination will be performed before IOP measurement or instillation of the fluorescein or lissamine green agent. Magnification will be consistent with standard clinical practice. The subject will be seated.

[00164] Slit lamp biomicroscopic observations will be graded as Normal or Abnormal. Abnormal findings will be categorized as clinically significant (findings that may interfere with study parameters or otherwise confound the data) or NCS. The following will be examined:

• Cornea

• Conjunctiva

• Anterior Chamber

• Iris

• Lens

• Eyelid

[00165] External magnification and biomicroscopy will be performed using a slitlamp. Magnification will be consistent with standard clinical practice. The subject will be seated.

[00166] IOP measurements will be performed in both eyes with a Goldmann applanation tonometer. The results will be recorded in mmHg. The tonometer should be checked for calibration accuracy according to the manufacturer’s instructions. Dilated fundus exams will be performed using indirect ophthalmoscopy. Observations of the vitreous, retina, macula, choroid and optic nerve will be made.

[00167] Schirmer’s tear test with topical anesthesia

[00168] The Schirmer’s test with topical anesthetic will be used to assess tear production using the following steps: 1. Topical anesthetic drops such as 0.5% proparacaine hydrochloride or equivalent should be instilled in both eyes of the subject. For Visit 3, 4, 5, 6, and 7 topical anesthetic drops should be instilled in both eyes of the subject once prior to the 30-minute Schirmer’s test and once prior to the 120-minute Schinner’s test.

2. The subject will be instructed to keep the eyes gently closed for one minute.

3. After opening the eyes and allowing the eyes to recover for approximately one additional minute, excess moisture in the inferior fornix is gently removed with a spear.

4. Schirmer’s strips (35 mm x 5 mm size filter paper strip) will be placed in each eye at the junction of the middle and lateral thirds of the lower eye lid.

5. Under ambient light, the subject will be instructed to look forward and to blink normally during the course of the test. The test should be performed in a room with no direct air or sunlight on the participant’s face.

6. The Schirmer’s strips should remain in place until five minutes have elapsed or both strips have reached maximum score.

7. After five minutes, strips will be removed from both eyes and the amount of wetting will be recorded.

[00169] Visual Analogue Scale-Symptom Index

[00170] The subjects will evaluate their symptoms (eye dryness, buming/stinging, itching, foreign body sensation, blurred vision, photophobia, pain) using a 100-point scale; 0=no discomfort and 100=maximal discomfort.

[00171] Subjects will be asked the following questions regarding ocular discomfort (unrelated to study drug instillation) at all visits.

[00172] The subject will be asked to rate each ocular symptom due to ocular dryness by placing a vertical mark on a horizontal line to indicate the current level of discomfort. 0% corresponds to “no discomfort” and 100% corresponds to “maximal discomfort.”

[00173] Tear break-up time (TBUT)

[00174] The examiner will instill 5 pL of 2% preservative-free sodium fluorescein solution into the inferior conjunctival cul-de-sac of each eye. To thoroughly mix the fluorescein with the tear film, the subject will be instructed to blink several times. In order to achieve maximum fluorescence, the examiner should wait approximately 30 seconds after instillation before evaluating TBUT.

[00175] With the aid of a slit-lamp, the examiner will monitor the integrity of the tear film, noting the time it takes to form micelles from the time that the eye is opened. TBUT will be measured in seconds using a stopwatch and a digital image recording system for the right eye followed by the left eye. A Wratten #12 yellow filter will be used to enhance the ability to grade TBUT.

[00176] Corneal and conjunctival staining

[00177] The 5 pL of 2% preservative-free sodium fluorescein solution that was instilled into the inferior conjunctival cul-de-sac of each eye for can also utilized for comeal and conjunctival staining assessments. A Wratten #12 yellow filter can be used to enhance the ability to grade fluorescein staining.

[00178] Following grading of the fluorescein staining, lissamine green dye will be instilled in each eye and the staining will be graded.

[00179] The following scale will be used to grade staining of the ocular surface (areas A, B, C, D, and E). Half (0.5) grade increments may be used.

[00180] Tear Film Osmolarity

[00181] Tear osmolarity will be measured in each eye using the TearLab© osmolarity system. At each time point, tear osmolarity will be taken once from the temporal canthus of each eye and the measurement will be recorded. A second reading may be taken if first reading is out of range. Tear osmolarity will be measured in mOsms/L.

[00182] Meibomian Gland Imaging and expression

[00183] Topical anesthetic drops such as 0.5% proparacaine hydrochloride or equivalent should be instilled in both eyes of the participant.

[00184] The meibomian glands in the tarsal plate of the lower eyelids will be examined by transillumination of the everted lid using slit-lamp microscopy. The glands will be examined for any altered anatomical features including gland drop out and abnormalities to the glandular orifices (plugging, pouting, narrowing, capping, cuffing, opaque/scarred).

[00185] A Mastrota paddle or forceps will be used to compress the inferior eyelid adnexa (lower, middle 5 glands) for approximately 5 to 15 seconds. Meibum will be graded according to the following scale, rating each eye separately. It should be documented if the glands cannot be expressed. [00186] Example 3

[00187] A. Methods

[00188] (a) Study Design

[00189] This was a four-week, single-center, double-masked, randomized, placebo- controlled, Phase II dry eye study evaluating the efficacy of Compound 1 0.034% solution compared to vehicle. The study consisted of three phases: a Screening Phase, a weekly Treatment Phase, and a BID Treatment Phase. The Screening Phase consisted of two visits (Visits 1 and 2) over a one to two-week period. Additional screening occurred at the beginning of Visit 3 prior to subject randomization and treatment. The weekly Treatment Phase occurred over a three-week period where dosing occurred at Visits 3, 4, 5, and 6. The BID Treatment Phase occurred over a one-week period and began at Visit 6 and continued until Visit 7. Dosing also occurred at Visit 7.

[00190] Eligible subjects were randomized using an interactive response system in a 1: 1 ratio to receive treatment with Compound 1 0.034% solution or Vehicle of Compound 1. Subjects received five in-office doses, and were instructed to instill one drop of the investigational product into each eye twice a day for one week.

[00191] (b) Screening and Eligibility

[00192] Screening occurred across three visits over a period of 1-2 weeks. To be eligible for the study, adult subjects (>18 years old) must have a history of dry eye in both eyes and used and/or desired to use artificial tears for DED symptoms within 6 months prior to Visit 1. Subjects were also required to meet the following key inclusion criteria: three anesthetized Schirmer’s tear tests (STTs) between 5-10mm at Visits 1, 2, and 3 in the study eye and no greater than 10mm in the companion eye; corneal fluorescein staining score >1 according to the Ora Calibra™ scale in at least one region in both eyes at Visits 1 and 2; a calculated best corrected visual acuity of 0.7 logarithm of the minimum angle of resolution (logMAR) or better in each eye as measured using an Early Treatment of Diabetic Retinopathy Study (ETDRS) chart.

[00193] All analyses were conducted based on the study eye, unless stated otherwise. The study eye was defined as the eye with the lower average of STT scores from Visits 1, 2. and 3 if three anesthetized STTs are between 5- 10mm. The remaining eye was considered the companion eye. If the average screening STT score was the same in both eyes, the right eye was selected as the study eye. [00194] Subjects were also prohibited from initiating, discontinuing, or changing dosage of a systemic medication known to cause ocular drying less than 14 days prior to Visit 1 (a change in dosage refers to any increase or decrease in the dose or any change in the dosing frequency). Subjects were not eligible for the study if they had an active ocular infection, ocular condition that could affect the subject’s safety or trial parameters, a surgical intervention within the previous 3 months, refractive surgery⁷ within the previous 6 months, or any significant illness. Subjects were prohibited from wearing contact or scleral lenses, using artificial tears or medications and treatments for dry eye and meibomian gland dysfunction, systemic of ocular antihistamines, lids scrubs, ocular and systemic antibiotics., immunosuppressants, and immunomodulators.

[00195] (c) Outcome Measures

[00196] Signs and symptoms of dry eye were assessed across all screening visits (Visit 1 and Visit 2) and all follow-up visits: Visit 3 (Day 1), Visit 4 (Day 8±1), Visit 5 (Day 15±1), Visit 6 (Day 22±1), and Visit 7 (Day 29±1). Signs were primarily assessed by anesthetized STT and investigator-rated comeal fluorescein staining (CFS), conducted at all visits. The STT was performed once at Visit 1 and 2. Additionally, at Visits 3-7, a STT was performed once 30-minutes prior to IP instillation and at 30, 35, 120, and 125 min post-IP instillation. The 35 and 125 min STTs measured tear replenishment dynamics and were performed immediately after the 30 and 120 min tests. Fluorescein staining of three areas of the comeal (superior, central, and inferior) was scored using the Ora Calibre™ scale (0-4 points with 0.5 increments permitted; 0= no staining to 4=confluent). Total comeal fluorescein staining (tCFS) was calculated as the sum of the total scores (maximum total score=12).

[00197] Subjects were asked to rate their eye dry ness and other symptoms at the beginning of each Visit using a series of questionnaires: the VAS, Ocular Surface Disease Index (OSDI), and ocular discomfort score (ODS). The VAS is a 7-item visual analogue scale assessing ocular symptoms due to ocular dryness based on the current level of discomfort (buming/stinging, itching foreign body sensation, blurred vision, photophobia, and pain on a 100-point scale [0=no discomfort, 100=maximal discomfort]). The OSDI is a 12-item questionnaire which asks the subject to assess symptoms experienced in the last week (symptoms [questions 1-5], visual-related function [6-9], and environmental triggers [questions 10-12]). The OSDI score is calculated on a scale of 0 to 100, with a larger score relating to greater severity. The ODS was graded by the subject rating each eye separately on a 5-point integer scale (0=no discomfort; 4=constant discomfort).

[00198] The primary endpoint was the improvement in STT (pre-IP instillation) at Visit 7 compared to baseline (Visit 3). Secondary efficacy endpoints assessed included improvement in STT, CFS, and symptom-based questionnaires (eye dryness score [EDS] of the VAS, OSDI, and ODS). STT secondary⁷ endpoints included improvement in STT (pre-IP instillation) at Visit 6 compared to baseline at Visit 3 and improvement in STT (pre-IP instillation) at Visit 7 compared to baseline at Visit 3 in the companion eye.

[00199] Additional exploratory assessments were conducted including conjunctival lissamine green staining, tear film break-up time (TBUT), meibum viscosity, and tear film osmolarity. Lissamine green staining of two areas of the conjunctiva (nasal and temporal) was scored using the Ora Calibra™ scale (0-4 points with 0.5 increments permitted; 0= no staining to 4=confluent). Total conjunctival lissamine green staining was calculated as the sum of the total scores (maximum total score=8).

[00200] Safety Assessments included adverse events (AEs), visual acuity, slit-lamp biomicroscopy, intraocular pressure, and dilated fundoscopy. Drop comfort of the IP was assessed for each eye upon instillation and at 1- and 2-minutes post-instillation while a descriptor query was conducted at 3 minutes post-instillation. Tolerability assessments occurred at all visits were IP-instillation occurred.

[00201] Treatment compliance during the BID dosing phase was determined by number of unopened investigational product vials returned and by reviewing patient dosing diaries. Subjects who missed >4 doses (consecutive and/or non-consecutive) in the correct w indow of time will be considered as non-compliant with BID dosing instructions. 100% of placebo-treated and 100% of Compound 1 subjects were compliant.

[00202] (d) Criteria for Evaluation:

[00203] Primary Efficacy Measures

[00204] Improvement in Schirmer’s tear test (STT) (pre-IP instillation) at Visit 7 compared to baseline (Visit 3).

[00205] Secondary Efficacy Measures

• Improvement in Schirmer’s tear test (STT) (pre-IP instillation) at Visit 7 compared to baseline (Visit 3) for companion eye.

• Improvement in Comeal Fluorescein Staining evaluated at Visit 3, 4, 5, 6 or 7. • Improvement in Ocular Surface Disease Index (OSDI) evaluated by the Subject at Visit 4, 5, 6 or 7.

• Improvement in Eye Dryness Score (EDS) of the Visual Analogue Scale (VAS) evaluated by the Subject at Visit 4, 5, 6 or 7.

• Improvement in the Ora Calibra™ Discomfort Score evaluated by the Subject at Visit 4. 5, 6 or 7.

• Improvement in Schirmer’s tear test (STT) (pre-IP instillation) at Visit 6 compared to baseline (Visit 3).

[00206] Exploratory Efficacy Measures:

• Replenishment Tear Dynamics as measured by STT at Visits 3, 4. 5, 6, and 7.

• Schirmer’s tear test (STT) (pre-IP instillation) at Visits 4 and 5.

• Conjunctival Staining assessed at all Visits.

• Tear Film Break-up Time (TBUT) assessed at all Visits.

• Digital photographs taken at all Visits.

• VAS Buming/Stinging evaluated by the Subject at all Visits.

• VAS Itching evaluated by the Subject at all Visits.

• VAS Foreign Body Sensation evaluated by the Subject at all Visits.

• VAS Blurred Vision evaluated by the Subject at all Visits.

• VAS Photophobia evaluated by the Subject at all Visits.

• VAS Pain evaluated by the Subject at all Visits.

• Meibomian Gland Imaging/Expression assessed at Visit 7.

• Tear Film Osmolarity assessed at Visit 7.

• Buccal Cell Samples Collected for Pharmacogenetic Assessments at Visit 3.

[00207] Safety Measures:

• Adverse Events assessed at all Visits.

• Visual Acuity at Distance Utilizing an ETDRS chart conducted at all Visits.

• Slit Lamp Biomicroscopy conducted at all Visits.

• Intraocular Pressure measured at Visits 1, 6, and 7.

• Dilated Fundoscopy measured at Visits 1, 6, and 7.

[00208] Tolerability Measures: • Drop comfort assessment (assessed by subjects upon instillation, and at 1- and 2-minutes post-instillation) at Visit 3, 4, 5,6, and 7. The drop comfort assessment results were summarized as continuous variable at eye level by treatment group with the Safety population.

• Drop descriptor query⁷ (assessed at 3 minutes post-instillation) at Visit 3, 4, 5, 6, and 7. The drop descriptor query results were summarized as categorical variables at eye level by treatment group with the Safety population.

[00209] (e) Statistical Methods

[00210] The primary analyses were performed on the ITT population with an analysis of covariance (ANCOVA) model for Schirmer’s tear test (STT) (pre-IP instillation) at Visit 7. Both the observed value and the change from baseline will be analyzed. The ANCOVA model will include baseline value as a covariate and treatment group as the main effect. The analysis was performed in the per protocol (PP) population.

[00211] The secondary endpoint of comeal fluorescein staining was analyzed in the same way as the primary endpoint. For remaining endpoints, treatment comparison was performed using Fisher’s exact test.

[00212] B. Interim Results

[00213] (a) Subject Disposition

[00214] Each of the 23 subjects enrolled in the study were randomized at Visit 3 to 1 of 2 treatment groups in a 1 : 1 ratio. In total, 11 subjects were assigned to receive Compound 1 (47.8%) and 12 subjects were assigned to receive placebo (52.2%).

[00215] The 23 randomized subjects received their first dose of assigned study drug in-office at Visit 3, thus comprising both the ITT population, the Per-Protocol population, and the Safety population.

[00216] A total of 20 subjects completed the study (n=10 and n=10 for placebo and Compound 1, respectively) and 3 subjects were discontinued (n=2 and n=l for placebo and Compound 1, respectively). Subjects’ age ranged from 50 to 88 years with and mean (SD) age of 68.7 (9.6). Of the 23 subjects, 56.52% were female subjects and 43.48% were males. The majority of subjects were white (95.65%).

[00217] (b) Primary Efficacy Endpoint

[00218] At baseline, the mean values for STT were 6.82mm/5min and 7.17mm/5 min for Compound 1 and placebo, respectively. Because of the length and other aspects of the design of the study, it was not possible to demonstrate a significant improvement in STT (pre-IP instillation) at Visit 7 (following three single weekly doses and 7 days of BID dosing) compared to baseline (Visit 3), with mean changes of 1.23mm and 1.47mm for Compound 1 and placebo, respectively (FIG. 2). To quantify the improvement in STT compared to baseline for Compound 1 and placebo requires further clinical investigation.

[00219] (c) Secondary Efficacy Endpoints

[00220] In the companion eye, the change of STT (pre-IP instillation) at Visit 7 compared to baseline (Visit 3) increased in both treatment groups; however, there was no statistically significant difference (FIG. 3A). No differences were observed in the change of STT (pre-IP instillation) at Visit 6 (following three single weekly doses) compared to baseline (Visit 3), with a mean change of -0.98mm and 0.62mm for Compound 1 and placebo, respectively. (FIG. 3B).

[00221] Surprisingly, when compared to pre-treatment screening Visit 2, Compound 1 treatment improved CFS at Visit 7 in the central region (p = 0.012), with a mean change of -0.13 and 0.93 for Compound 1 and placebo groups, respectively. Both groups saw an increase in total comeal fluorescein staining (tCFS). with the Compound 1 group increasing less (1.33 and 2.87; p = 0.024) (FIG. 4A). No significant differences were observed in the companion eye, although similar trends were observed as in study eye (FIG. 4B).

[00222] Similarly, when compared to the post-first dose assessment at Visit 3, Compound 1 improved CFS in the central region and in the tCFS score at Visit 7, with a mean change of -0.65 and 0.20 [p=0.02] in central regions and -0.91 and 0.36 [p=0.015] in total score for Compound 1 and placebo groups, respectively. The Compound 1 group exhibited a numerical CFS score decrease in superior and inferior regions, whereas the placebo group demonstrated a numerical increase; however, the duration and other aspects of the design of the study did not produce differences that were statistically significant (FIG. 5A). Compound 1 -treated subjects showed numerically greater CFS improvement in all regions in the companion eye at Visit 7 from Visit 3, the duration and other aspects of the design of the study did not result in these differences reaching statistical significance (FIG. 5B). To better quantify the improvements in CFS following Compound 1 administration, as compared to placebo, additional studies are required.

[00223] For all key symptom-based assessments, both treatment groups showed improvement trends in EDS of the VAS, OSDI, and ODS, at Visit 7 compared to baseline (Visit 3). Because of the length and other aspects of the design of the study, it was not possible to demonstrate a significant difference between treatment groups (FIG. 6). To better quantify⁷ improvements in key symptoms following administration of Compound 1, as compared to placebo requires further clinical investigation.

[00224] (d) Exploratory Measures

[00225] No statistically significant differences were observed between Compound 1 and placebo groups in tear dynamics as measured by STT at 30, 35, 120, 125 minutes (min) post-IP instillation at any visit; with the exception of a greater change from Visit baseline in the placebo group at the 120 min measurement at Visit 7 (p=0.037). Numerical trends were observed in tear dynamics across study visits. At Visits 3.4 and 5, subjects treated with Compound 1 had numerically higher mean change in STT 30 min post-IP instillation from Visit baseline compared to placebo (FIG. 7). This effect was not observed at Visit 6 or 7 or at any 120 min time point post drug administration. The 35- and 125-min STTs, which assess replenishment tear dynamics, were generally numerically higher in subjects treated with Compound 1, without reaching statistical significance (FIG. 8). No differences were observed between groups in change from baseline in STT (pre-IP) at Visit 4 and Visit 5, after one and two single weekly doses, respectively (FIG. 9).

[00226] Conjunctival lissamine green staining increased in the Compound 1 and placebo groups, at Visit 7 from pre-treatment screening Visit 2. in the in the nasal and temporal conjunctival regions and in the total score with no statistically significant difference between treatments (FIG. 10A). In the companion eye, both groups saw an increase in total score from Visit 2, but the increase observed in the Compound 1 group was significantly smaller than in the placebo group (0.81 and 2.14; p = 0.043) (FIG. 10B). When compared to the post-first dose assessment at Visit 3, numerical improvement was observed in the nasal and temporal regions and in the total score at Visit 7 in the Compound 1 -treated group; how ever the effect did not reach statistical significance (FIG. 11 A). A similar trend w as observed in the companion eye (FIG. 1 IB).

[00227] All other symptom parameters demonstrated general improvement at the final Visit (Visit 7) from baseline (Visit 3); how ever no statistically significant differences were observed between groups for blurred vision, foreign body sensation, buming/stinging, itching, pain, and photophobia (FIG. 12). The analysis of tear break-up time (Figure 13), tear osmolarity (FIG. 14), and meibum appearance (FIG. 15) showed no significant differences.

[00228] (e) Safety

[00229] Among randomized subjects, there were a total of 22 AEs reported across 12 subjects (Table 2). 5 subjects experienced 9 AEs in the Compound 1 group while 7 subjects of the placebo group experienced 13 AEs. The most common AE (>20% incidence in each group) was burning upon IP instillation in the Compound 1 group and burning upon IP instillation and decrease in visual acuity in the placebo group. All were mild or moderate in severity. Visual acuity, slit lamp biomicroscopy, dilated fundoscopy measurements and IOP measurements did not indicate any notable safety concerns for Compound 1 ophthalmic solution.

[00230] C. Discussion

[00231] For the secondary endpoint of CFS, Compound 1 improved staining in the central region and in the tCFS from Visit 3 to 7, after completion of the BID dosing regimen. Compound 1 treatment improved staining in the central region from Visit 2 to 7 while both treatment groups showed an increase in tCFS, with the Compound 1 group increasing less.

[00232] The improvement in CFS staining was most pronounced when comparing the final visit (Visit 7) to Visit 3 (after initial dose), likely due to the repeated administration of anesthetized STT in this dry eye population. At Visit 2, staining was assessed after one anesthetized STT, while at Visits 3 to 7 staining was scored after five STTs and instillation of anesthetic three times, over a 3 -hour period. While no immediate drug effect on staining was expected after a single dose, the cumulative impact of the STTs in this dry eye population likely influenced staining; supported by the increase in staining scores from Visit 2 to Visit 3. Despite this, improvement was still observed in the central region at Visit 2 to 7 in the Compound 1 treated group.

[00233] Because of the duration and other aspects of the design of the studies, the differences observed in symptom-based secondary endpoints (EDS, OSDI. and Ocular Discomfort Score) did not reach statistical significance, necessitating additional clinical investigations to better quantity⁷ those differences.

[00234] Compound 1 was well tolerated, and reported AEs were mild or moderate in severity. No changes in visual acuity, slit lamp biomicroscopy, dilated fundoscopy measurements and IOP measurements were detected. [00235] Overall, these results suggest that Compound 1 induces rapid decreases in comeal staining and support the design of more definitive clinical investigations to better demonstrate Compound 1’s therapeutic effects on dry eye symptoms.

[00236] The references cited are expressly incorporated by reference herein in their entireties.

Claims

What is claimed is:

1. A method of treating dry eye disease in a human in need of treatment thereof comprising instilling once weekly, into the eye of the human, a drop of an ophthalmic solution comprising between 0.03% and 0.05% w/v of Compound 1, or a pharmaceutically acceptable acid addition salt thereof:

Compound 1.

2. The method of claim 1, wherein the treatment results in an improvement in at least one sign of dry eye disease in the human.

3. The method of any one of claim 1 or claim 2, wherein the treatment results in an improvement in at least one symptom of dry' eye disease in the human.

4. The method of any one of the preceding claims, wherein the treatment results in an improvement in the eye of the human of one or more of eye dryness, burning, stinging, itching, foreign body sensation, blurred vision, photophobia, or pain.

5. The method of any one of the preceding claims, wherein the dry eye disease is aqueous deficient dry eye, evaporative dry eye, or mixed dry eye.

6. The method of any one of the preceding claims, wherein the human has Sjogren’s syndrome.

7. The method of any one of the preceding claims, wherein the dry eye disease in the human is associated with aging, contact lens usage, or refractive surgery.

8. The method of any one of the preceding claims, wherein the dry eye disease in the human is associated with a systemic immunologic disease.

9. The method of any one of the preceding claims, wherein the dry eye disease is definite dry eye disease or probable dry eye disease.

10. The method of any one of the preceding claims, wherein the human has an anesthetized Schirmer's tear test (STT) of between 5 and 10 mm/5 min in the eye at baseline.

11. The method of any one of the preceding claims, wherein the method results in an improvement in the human’s anesthetized Schirmer's tear test (STT), as compared to baseline.

12. The method of any one of the preceding claims, wherein the method results in an improvement in Corneal Fluorescein Staining, as compared to baseline.

13. The method of any one of the preceding claims, wherein the method results in an improvement in the human’s self-reported ocular discomfort, as compared to baseline.

14. The method of any one of the preceding claims, wherein the method results in an improvement in the human’s Eye Dryness Score (EDS) of the Visual Analogue Scale (VAS), as compared to baseline.

15. The method of any one of the preceding claims, wherein the method results in an improvement in the human’s self-reported ocular irritation, as compared to baseline.

16. The method of any one of the preceding claims, wherein the method results in an improvement in meibomian gland secretion viscosity, as compared to baseline.

17. The method of any one of the preceding claims, wherein the human has a calculated best-corrected visual acuity of at least about 0.7 logarithm of the minimum angle of resolution (logMAR) in the eye at baseline.

18. The method of any one of the preceding claims, wherein the human has a comeal fluorescein staining in at least one region of the eye, at baseline.

19. The method of any one of the preceding claims, wherein the dry eye disease is characterized by one or more of visual disturbances, changes in tears, epithelial damage, unstable tear film, ocular photophobia, ocular foreign body sensation, ocular itching, ocular grittiness, ocular burning, or conjunctival redness.

20. The method of any one of the preceding claims, wherein the human is 18 years of age or older.

21. The method of any one of the preceding claims, wherein the ophthalmic solution comprises a pharmaceutically acceptable acid addition salt of Compound 1.

22. The method of any one of the preceding claims, wherein the ophthalmic solution comprises Compound 1.

23. The method of any one of the preceding claims, wherein the ophthalmic solution comprises 0.034% w/v of Compound 1.

24. The method of any one of the preceding claims, wherein the ophthalmic solution comprises:

25. The method of any one of the preceding claims, wherein the drop of the ophthalmic solution is instilled once weekly, unilaterally, into the eye of the human.

26. The method of any one of the preceding claims, wherein the drop of the ophthalmic solution is instilled once weekly, bilaterally, into the eyes of the human.

27. The method of any one of the preceding claims, wherein the method is performed for a treatment period of at least one week.

28. The method of any one of the preceding claims, wherein the method is performed for a treatment period of at least two weeks.

29. The method of any one of the preceding claims, wherein the method is performed for a treatment period of at least three weeks.

30. The method of any one of claims 26-28, wherein after the treatment period, the drop of the ophthalmic solution is instilled twice daily, into the eye of the human.

31. The method of claim 29, wherein the twice daily instillation is performed unilaterally, into the eye of the human.

32. The method of claim 29, wherein the twice daily instillation is performed bilaterally into the eyes of the human.

33. A method of treating dry eye disease in a human in need of treatment thereof, comprising instilling twice daily, into the eye of the human, a drop of an ophthalmic solution comprising between 0.03% and 0.05% w/v of Compound 1, or a pharmaceutically acceptable acid addition salt thereof:

Compound 1.