Methods and compositions for increasing pupil dilation

Abstract

Provided are methods and compositions for increasing a pupil measurement.

Description

METHODS AND COMPOSITIONS FOR INCREASING PUPIL DILATIONCROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 574,098, filed April 3, 2024, which is incorporated herein by reference in its entirety.SUMMARY

[0002] In certain aspects, described herein is a method of identifying a formulation suitable for treatment of premyopia or myopia, the method comprising administering the formulation that increases the diameter of the pupil of the eye by at least 10% for a period of 60 minutes or more. In some embodiments, the diameter of the pupil is measured by pupillometry. In some embodiments, the diameter is increased by at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90 % for a period of 60 minutes or more. In some embodiments, the diameter is increased for a period of 90 minutes or more. In some embodiments, the eye is a mammalian eye. In some embodiments, the eye is a human eye. In some embodiments, the eye is a rabbit eye. In some embodiments, the eye is an adult human eye. In some embodiments, the eye is a pediatric human eye. In some embodiments, the pupil diameter prior to administration is from about 2 mm to about 7 mm. In some embodiments, the pupil diameter prior to administration is from about 2 mm to about 8 mm. In some embodiments, the formulation is administered to only one eye of the subject. In some embodiments, the formulation is administered to both eyes of the subject. In some embodiments, the formulation is compared to a reference formulation. In some embodiments, the reference formulation comprises D2O. In some embodiments, the reference formula comprises BAK. In some embodiments, the reference formula has a pD of about 7 or a pH of about 6.6. In some embodiments, the reference formula comprises BAK, is formulated in D2O, and has a pD of about 7. In some embodiments, the reference formulation comprises the same concentration of atropine as the formulation. In some embodiments, the formulation is identified as suitable for treatment of myopia or premyopia when the formulation increases the diameter of the pupil of the eye by at least 10% for a period of 60 minutes or more. In some embodiments, the method comprises calculating the area under the curve (AUC) of the pupil diameter for 12 hours for the formulation and the reference formulation. In some embodiments, the method comprises calculating the area under the curve (AUC) of the pupil diameter for 24hours for the formulation and the reference formulation. In some embodiments, the formulation is identified as suitable for treatment of myopia or premyopia when the AUC of the formulation is at least 50% of the reference formulation for a period of 60 minutes or more, wherein the reference formulation comprises D2O and the formulation does not comprise D2O.

[0003] In certain aspects, described herein is a method of identifying a formulation suitable for treatment of premyopia or myopia, the method comprising: administering a formulation to an eye of the subject; and measuring the diameter of the pupil of the eye before administration and at a timepoint at least 60 minutes after administration. In some embodiments, the diameter of the pupil is measured by pupillometry. In some embodiments, the diameter of the pupil is measured at two or more timepoints after administration. In some embodiments, the diameter is increased by at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90 % for a period of 60 minutes or more. In some embodiments, the diameter is increased for a period of 90 minutes or more. In some embodiments, the eye is a mammalian eye. In some embodiments, the eye is a human eye. In some embodiments, the eye is a rabbit eye. In some embodiments, the eye is an adult human eye. In some embodiments, the eye is a pediatric human eye. In some embodiments, the pupil diameter prior to administration is from about 2 mm to about 7 mm. In some embodiments, the pupil diameter prior to administration is from about 2 mm to about 8 mm. In some embodiments, the formulation is administered to only one eye of the subject. In some embodiments, the formulation is administered to both eyes of the subject. In some embodiments, the formulation is compared to a reference formulation. In some embodiments, the reference formulation comprises D2O. In some embodiments, the reference formula comprises BAK. In some embodiments, the reference formula has a pD of about 7 or a pH of about 6.6. In some embodiments, the reference formula comprises BAK, is formulated in D2O, and has a pD of about 7. In some embodiments, the reference formulation comprises the same concentration of atropine as the formulation. In some embodiments, the formulation is identified as suitable for treatment of myopia or premyopia when the formulation increases the diameter of the pupil of the eye by at least 10% for a period of 60 minutes or more. In some embodiments, the method comprises calculating the area under the curve (AUC) of the pupil diameter for 12 hours for the formulation and the reference formulation. In some embodiments, the method comprises calculating the area under the curve (AUC) of the pupil diameter for 24 hours for the formulation and the reference formulation. In some embodiments, the formulation is identified as suitable for treatment of myopia or premyopia when the AUC of the formulation is at least 50% of the reference formulation for a period of 60 minutes or more, wherein the reference formulation comprises D2O and the formulation does not comprise D2O.

[0004] A method of assessing potency of a water-based low-dose atropine formulation by administering a drop of said formulation to the eye of a subject and measuring the diameter of a pupil of an eye for a period of 60 minutes or more and comparing said formulation to a second reference formulation of an identical concentration of low-dose atropine also comprising D2O and BAK.

[0005] A method of increasing the diameter of a pupil of an eye by at least 10% for a period of 60 minutes or more, comprising administering a formulation comprising atropine, D2O, and BAK to the eye. In some embodiments, the formulation comprises a pD of about 7. In some embodiments, the diameter is increased relative to a reference formulation comprising atropine at the same dose, wherein the formulation does not comprise BAK. In some embodiments, the reference formulation comprises a pD of less than about 4 or a pH of less than or about 3.6. In some embodiments, the diameter is increased by at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90 % for a time period of 60 minutes or more. In some embodiments, the diameter is increased for a time period of 90 minutes or more. In some embodiments, the eye is a mammalian eye. In some embodiments, the eye is a human eye. In some embodiments, the eye is an adult human eye. In some embodiments, the eye is a pediatric human eye. In some embodiments, the pupil diameter prior to administration is from about 2mm and to about 7 mm. In some embodiments, the pupil diameter prior to administration is from about 2 mm and to about 8 mm. In some embodiments, the formulation is administered to only one eye of the subject. In some embodiments, the formulation is administered to both eyes of the subject. In some embodiments, the atropine is present in the ophthalmic composition at a concentration of one of: from about 0.001 wt% to about 0.40 wt%, from about 0.001 wt% to about 0.30 wt%, from about 0.001 wt% to about 0.20 wt%, from about 0.001 wt% to about 0. 10 wt%, from about 0.001 wt% to about 0.09 wt%, from about 0.001 wt% to about 0.08 wt%, from about 0.001 wt% to about 0.07 wt%, from about 0.001 wt% to about 0.06 wt%, from about 0.001 wt% to about 0.05 wt%, from about 0.001 wt% to about 0.04 wt%, from about 0.001 wt% to about 0.03 wt%, from about 0.001 wt% to about 0.025 wt%, from about 0.001 wt% to about 0.02 wt%, from about 0.001 wt% to about 0.01 wt%, from about 0.001 wt% to about 0.008 wt%, or from about 0.001 wt% to about 0.005 wt%. In some embodiments, the atropine is at a concentration of 0.001 wt% to 0. 1 wt%. In some embodiments, the ophthalmic composition further comprises an osmolarity adjusting agent. In some embodiments, the osmolarity adjusting agent is sodium chloride. In some embodiments, the sodium chloride is present in the ophthalmic composition at a concentration of one of: from about 0.01 wt% to about 1.0 wt%, from about 0.05 wt% to about 1.5 wt%, from about 0.075 wt% to about 2.0 wt%, or from about 0. 1 wt% to about 3.0 wt%. Insome embodiments, the ophthalmic composition further comprises a buffering agent. In some embodiments, the buffering agent comprises citrate.

[0006] A method of increasing the diameter of a pupil of an eye by at least 10% for a period of 60 minutes or more, comprising administering a formulation comprising atropine, and BAK to the eye, wherein the composition comprises a pD of about 7 or a pH of about 6.6. In some embodiments, the composition does not comprise D2O. In some embodiments, the composition comprises D2O. In some embodiments, the diameter is increased relative to a reference formulation comprising atropine at the same dose, wherein the reference formulation does not comprise BAK. In some embodiments, the reference formulation comprises a pD of less than about 4 or a pH of less than about 3.6. In some embodiments, the diameter is increased by at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90 % for a time period of 60 minutes or more. In some embodiments, the diameter is increased for a time period of 90 minutes or more. In some embodiments, the eye is a mammalian eye. In some embodiments, the eye is a human eye. In some embodiments, the eye is an adult human eye. In some embodiments, the eye is a pediatric human eye. In some embodiments, the pupil diameter prior to administration is between 2mm and 7 mm. In some embodiments, the pupil diameter prior to administration is between 2 mm and 8 mm. In some embodiments, the formulation is administered to only one eye of the subject. In some embodiments, the formulation is administered to both eyes of the subject. In some embodiments, the atropine is present in the ophthalmic composition at a concentration of one of: from about 0.001 wt% to about 0.40 wt%, from about 0.001 wt% to about 0.30 wt%, from about 0.001 wt% to about 0.20 wt%, from about 0.001 wt% to about 0.10 wt%, from about 0.001 wt% to about 0.09 wt%, from about 0.001 wt% to about 0.08 wt%, from about 0.001 wt% to about 0.07 wt%, from about 0.001 wt% to about 0.06 wt%, from about 0.001 wt% to about 0.05 wt%, from about 0.001 wt% to about 0.04 wt%, from about 0.001 wt% to about 0.03 wt%, from about 0.001 wt% to about 0.025 wt%, from about 0.001 wt% to about 0.02 wt%, from about 0.001 wt% to about 0.01 wt%, from about 0.001 wt% to about 0.008 wt%, or from about 0.001 wt% to about 0.005 wt%. In some embodiments, the atropine is at a concentration of 0.001 wt% to 0.1 wt%. In some embodiments, the ophthalmic composition further comprises an osmolarity adjusting agent. In some embodiments, the osmolarity adjusting agent is sodium chloride. In some embodiments, the sodium chloride is present in the ophthalmic composition at a concentration of one of: from about 0.01 wt% to about 1.0 wt%, from about 0.05 wt% to about 1.5 wt%, from about 0.075 wt% to about 2.0 wt%, or from about 0.1 wt% to about 3.0 wt%. In some embodiments, the ophthalmic composition further comprises a buffering agent. In some embodiments, the buffering agent comprises citrate.INCORPORATION BY REFERENCE

[0007] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BRIEF DESCRIPTION OF THE DRAWINGS

[0008] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

[0009] FIG. 1A illustrates a comparison of rabbit pupil diameters following administration of TA2 (0.02 wt% Atropine, D2O, and 0.01% BAK, 5 mM citrate, 0.9% NaCl) and TA3 (0.02 wt% Atropine, H2O, 5 mM citrate, 0.9% NaCl, and no BAK), respectively, across multiple timepoints. FIG. IB depicts the comparison of rabbit pupil diameters, extrapolated to 24 hours.

[0010] FIGS. 2A-2D depict comparisons of rabbit pupil diameters relative to baseline values following administration TA2 (0.02 wt% Atropine, D2O, and 0.01% BAK, 5 mM citrate, 0.9% NaCl) and TA3 (0.02 wt% Atropine, H2O, 5 mM citrate, 0.9% NaCl, and no BAK), respectively across multiple timepoints. FIG. 2A and FIG. 2B depict comparisons of right eye (FIG. 2A) and left eye (FIG. 2B) pupil diameter measurements relative to baseline values in male rabbits. FIG. 2C and FIG. 2D depict comparisons of right eye (FIG. 2C) and left eye (FIG. 2D) pupil diameter measurements relative to baseline values in female rabbits.

[0011] FIG. 3 depicts the experimental protocol.DETAILED DESCRIPTION

[0012] Described herein are ophthalmic compositions and methods for evaluating the efficacy of ophthalmic compositions. One challenge associated with developing ophthalmic compositions, such as low-dose atropine formulations as described herein, is ensuring effective transport of the drug into the eye. Pupil diameter may be used as a surrogate marker for the amount of atropine transport across the cornea into the eye. Thus, pupil diameter, and related measurements may increase as increased concentrations of atropine are transported into the eye.

[0013] Myopia, commonly known as nearsightedness, is a refractive error where distant objects appear blurry while close objects can be seen clearly. Myopia occurs when the eyeball is too long relative to the focusing power of the cornea and lens, causing light rays to focus in front of the retina. Myopia can be classified by severity: mild myopia (up to -3.00 diopters), moderate myopia (-3.00 to -6.00 diopters), and high myopia (greater than -6.00 diopters). Premyopia can refer to the early stages of myopia development, often seen in children or adolescents who may exhibit symptoms but have not yet been diagnosed with myopia.

[0014] Clinicians can assess various parameters to tailor ophthalmic disorder management strategies, including pupil size, baseline refraction, axial length measurements, age of onset, family history, and lifestyle factors. Regular monitoring of these parameters can allow for adjustments in treatment strategies to achieve optimal outcomes.I. METHODS

[0015] Provided herein are methods and compositions for increasing a measurement, such as a pupil measurement, comprising administering a formulation described herein to the eye. Also described herein are methods for identifying a formulation of an ophthalmic composition suitable for treating myopia or premyopia. In some embodiments, the method comprises administering a formulation that increases a measurement for a time period. The measurement may be a pupil diameter. The measurement may be the area under the curve of the pupil diameter over 12 or 24 hours. In some embodiments, described herein is a method of assessing potency of a water-based low-dose atropine formulation administering one or more drops of said formulation to the eye of the subject, and measuring the diameter of the pupil of the eye for a period of 60 minutes or more or more after administration, and comparing measurements obtained from said formulation to a second reference formulation comprising an identical concentration of low-dose atropine, and further comprising D2O and BAK. In some embodiments, the diameter of the pupil is measured prior to administration.

[0016] In some embodiments, the composition is formulated in H2O and comprises a pH. In some instances, the composition is formulated in D2O and comprises a pD. An equivalent pD may be calculated from a pH by adding 0.4. For instance, a pH of 6.6 is equivalent to a pD of 7.0.

[0017] In some embodiments, the formulation is an ophthalmic composition as described herein. In some embodiments, the formulation comprises D2O, BAK and atropine. In some embodiments, the formulation comprises a pD of about 7. In some embodiments, the formulation comprises a pD of about 7.4. In some embodiments, the formulation comprises a pDof about 6 to about 8. In some embodiments, the formulation comprises a pD of about 6 to about6.2, about 6 to about 6.4, about 6 to about 6.6, about 6 to about 6.8, about 6 to about 7, about 6 to about 7.2, about 6 to about 7.6, about 6 to about 7.8, about 6 to about 8, about 6.2 to about 6.4, about 6.2 to about 6.6, about 6.2 to about 6.8, about 6.2 to about 7, about 6.2 to about 7.2, about 6.2 to about 7.6, about 6.2 to about 7.8, about 6.2 to about 8, about 6.4 to about 6.6, about 6.4 to about 6.8, about 6.4 to about 7, about 6.4 to about 7.2, about 6.4 to about 7.6, about 6.4 to about 7.8, about 6.4 to about 8, about 6.6 to about 6.8, about 6.6 to about 7, about 6.6 to about7.2, about 6.6 to about 7.6, about 6.6 to about 7.8, about 6.6 to about 8, about 6.8 to about 7, about 6.8 to about 7.2, about 6.8 to about 7.6, about 6.8 to about 7.8, about 6.8 to about 8, about 7 to about 7.2, about 7 to about 7.6, about 7 to about 7.8, about 7 to about 8, about 7.2 to about7.6, about 7.2 to about 7.8, about 7.2 to about 8, about 7.6 to about 7.8, about 7.6 to about 8, or about 7.8 to about 8. In some embodiments, the formulation comprises a pD of about 6, about6.2, about 6.4, about 6.6, about 6.8, about 7, about 7.2, about 7.6, about 7.8, or about 8. In some embodiments, the formulation comprises a pD of at least about 6, about 6.2, about 6.4, about6.6, about 6.8, about 7, about 7.2, about 7.6, or about 7.8. In some embodiments, the formulation comprises a pD of at most about 6.2, about 6.4, about 6.6, about 6.8, about 7, about 7.2, about7.6, about 7.8, or about 8.

[0018] In some embodiments, the formulation comprises H2O, BAK, and atropine. In some embodiments, the formulation comprises a pH of about 6.6. In some embodiments, the formulation comprises a pH of about 7. In some embodiments, the formulation comprises a pH of about 5.6 to about 7.6. In some embodiments, the formulation comprises a pH of about 5.6 to about 5.8, about 5.6 to about 6, about 5.6 to about 6.2, about 5.6 to about 6.4, about 5.6 to about 6.8, about 5.6 to about 6.8, about 5.6 to about 7.2, about 5.6 to about 7.4, about 5.6 to about 7.6, about 5.8 to about 6, about 5.8 to about 6.2, about 5.8 to about 6.4, about 5.8 to about 6.6, about 5.8 to about 6.8, about 5.8 to about 7.2, about 5.8 to about 7.4, about 5.8 to about 7.6, about 6 to about 6.2, about 6 to about 6.4, about 6 to about 6.6, about 6 to about 6.8, about 6 to about 7.2, about 6 to about 7.4, about 6 to about 7.6, about 6.2 to about 6.4, about 6.2 to about 6.8, about 6.2 to about 6.8, about 6.2 to about 7.2, about 6.2 to about 7.4, about 6.2 to about 7.6, about 6.4 to about 6.6, about 6.4 to about 6.8, about 6.4 to about 7.2, about 6.4 to about 7.4, about 6.4 to about 7.6, about 6.6 to about 6.8, about 6.6 to about 7.2, about 6.6 to about 7.4, about 6.6 to about 7.6, about 6.8 to about 7.2, about 6.8 to about 7.4, about 6.8 to about 7.6, about 7.2 to about 7.4, about 7.2 to about 7.6, or about 7.4 to about 7.6. In some embodiments, the formulation comprises a pH of about 5.6, about 5.8, about 6, about 6.2, about 6.4, about 6.6, about 6.8, about 7.2, about 7.4, or about 7.6. In some embodiments, the formulation comprises apH of at least about 5.6, about 5.8, about 6, about 6.2, about 6.4, about 6.6, about 6.8, about 7.2, or about 7.4. In some embodiments, the formulation comprises a pH of at most about 5.8, about 6, about 6.2, about 6.4, about 6.6, about 6.8, about 7.2, about 7.4, or about 7.6.

[0019] In some embodiments, the measurement is increased relative to a reference formulation comprising atropine at the same dose. In some embodiments, the reference formulation does not comprise BAK. In some embodiments, the reference formulation comprises a pH of about 3.6.In some embodiments, the reference formulation comprises a pD of about 4. In some embodiments, the reference formulation comprises H2O. In some embodiments, the reference formulation comprises D2O. In some embodiments, the reference formulation comprises a pH of about 6.6. In some embodiments, the reference formulation comprises a pD of about 7.

[0020] In some embodiments, the reference formulation comprises a pD of about 4 to about 8. In some embodiments, the reference formulation comprises a pD of about 4 to about 4.5, about 4 to about 5, about 4 to about 5.5, about 4 to about 6, about 4 to about 6.5, about 4 to about 7, about 4 to about 7.5, about 4 to about 8, about 4.5 to about 5, about 4.5 to about 5.5, about 4.5 to about 6, about 4.5 to about 6.5, about 4.5 to about 7, about 4.5 to about 7.5, about 4.5 to about 8, about 5 to about 5.5, about 5 to about 6, about 5 to about 6.5, about 5 to about 7, about 5 to about 7.5, about 5 to about 8, about 5.5 to about 6, about 5.5 to about 6.5, about 5.5 to about 7, about 5.5 to about 7.5, about 5.5 to about 8, about 6 to about 6.5, about 6 to about 7, about 6 to about 7.5, about 6 to about 8, about 6.5 to about 7, about 6.5 to about 7.5, about 6.5 to about 8, about 7 to about 7.5, about 7 to about 8, or about 7.5 to about 8. In some embodiments, the reference formulation comprises a pD of about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, or about 8. In some embodiments, the reference formulation comprises a pD of at least about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, or about 7.5. In some embodiments, the reference formulation comprises a pD of at most about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, or about 8.

[0021] In some embodiments, the reference formula comprises a pH of about 3.6 to about 7.6. In some embodiments, the reference formula comprises a pH of about 3.6 to about 4.1, about 3.6 to about 4.6, about 3.6 to about 5.1, about 3.6 to about 5.6, about 3.6 to about 6.1, about 3.6 to about 6.6, about 3.6 to about 7.1, about 3.6 to about 7.6, about 4.1 to about 4.6, about 4.1 to about 5.1, about 4.1 to about 5.6, about 4.1 to about 6.1, about 4.1 to about 6.6, about 4.1 to about 7.1, about 4.1 to about 7.6, about 4.6 to about 5.1, about 4.6 to about 5.6, about 4.6 to about 6.1, about 4.6 to about 6.6, about 4.6 to about 7.1, about 4.6 to about 7.6, about 5.1 to about 5.6, about 5.1 to about 6.1, about 5.1 to about 6.6, about 5.1 to about 7.1,about 5.1 to about 7.6, about 5.6 to about 6.1, about 5.6 to about 6.6, about 5.6 to about 7.1, about 5.6 to about 7.6, about 6.1 to about 6.6, about 6.1 to about 7.1, about 6.1 to about 7.6, about 6.6 to about 7.1, about 6.6 to about 7.6, or about 7.1 to about 7.6. In some embodiments, the reference formula comprises a pH of about 3.6, about 4.1, about 4.6, about 5.1, about 5.6, about 6.1, about 6.6, about 7.1, or about 7.6. In some embodiments, the reference formula comprises a pH of at least about 3.6, about 4.1, about 4.6, about 5.1, about 5.6, about 6.1, about 6.6, or about 7. 1. In some embodiments, the reference formula comprises a pH of at most about 4.1, about 4.6, about 5.1, about 5.6, about 6.1, about 6.6, about 7.1, or about 7.6.

[0022] In certain aspects, described herein is a method of increasing the diameter of a pupil of an eye by at least 10% for a period of 60 minutes or more, comprising administering a formulation comprising atropine, D2O, and BAK to the eye. In certain aspects, described herein is a method of increasing the diameter of a pupil of an eye by at least 10% for a period of 60 minutes or more, comprising administering a formulation comprising atropine and BAK to the eye, wherein the composition comprises a pD of about 7.

[0023] In certain aspects, described herein is a method of identifying a formulation suitable for treatment of premyopia or myopia, the method comprising administering the formulation that increases the diameter of the pupil of the eye by at least 10% for a period of 60 minutes or more. In certain aspects, described herein is a method of identifying a formulation suitable for treatment of premyopia or myopia, the method comprising administering a formulation to an eye of the subject; and measuring the diameter of the pupil of the eye before administration and at a timepoint at least 60 minutes after administration. In some embodiments, the formulation is compared to a reference formulation.A. Pupil measurements

[0024] The methods and compositions described herein can result in alterations to a measurement of an eye, such as a pupil diameter measurement. The pupil diameter measurement may provide a baseline measurement prior to undergoing a treatment via administration of an ophthalmic formulation. In some aspects, the pupil diameter measurement can inform assessment of an ocular response to the treatment before and after administration, progression of an ophthalmic condition (e.g., myopia), identification of a suitable treatment regimen, identification of adverse effects in response to the treatment, or a combination thereof.

[0025] In some embodiments, the eye is an adult eye. In some embodiments, the eye is a pediatric eye. In some embodiments, the pupil diameter prior to administration is about 2 mm to about 8 mm. In some embodiments, the pupil diameter prior to administration is about 2 mm toabout 4 mm. In some embodiments, the pupil diameter prior to administration is about 2 mm to about 3 mm, about 2 mm to about 4 mm, about 2 mm to about 5 mm, about 2 mm to about 6 mm, about 2 mm to about 7 mm, about 2 mm to about 8 mm, about 3 mm to about 4 mm, about3 mm to about 5 mm, about 3 mm to about 6 mm, about 3 mm to about 7 mm, about 3 mm to about 8 mm, about 4 mm to about 5 mm, about 4 mm to about 6 mm, about 4 mm to about 7 mm, about 4 mm to about 8 mm, about 5 mm to about 6 mm, about 5 mm to about 7 mm, about 5 mm to about 8 mm, about 6 mm to about 7 mm, about 6 mm to about 8 mm, or about 7 mm to about 8 mm. In some embodiments, the pupil diameter prior to administration is about 2 mm, about 3 mm, about 4 mm, about 5 mm, about 6 mm, about 7 mm, or about 8 mm. In some embodiments, the pupil diameter prior to administration is at least about 2 mm, about 3 mm, about 4 mm, about 5 mm, about 6 mm, or about 7 mm. In some embodiments, the pupil diameter prior to administration is at most about 3 mm, about 4 mm, about 5 mm, about 6 mm, about 7 mm, or about 8 mm.

[0026] In some embodiments, the pupil diameter after administration is about 2 mm to about 8 mm. In some embodiments, the pupil diameter after administration is about 4 mm to about 8 mm. In some embodiments, the pupil diameter after administration is about 2 mm to about 3 mm, about 2 mm to about 4 mm, about 2 mm to about 5 mm, about 2 mm to about 6 mm, about 2 mm to about 7 mm, about 2 mm to about 8 mm, about 3 mm to about 4 mm, about 3 mm to about 5 mm, about 3 mm to about 6 mm, about 3 mm to about 7 mm, about 3 mm to about 8 mm, about 4 mm to about 5 mm, about 4 mm to about 6 mm, about 4 mm to about 7 mm, about4 mm to about 8 mm, about 5 mm to about 6 mm, about 5 mm to about 7 mm, about 5 mm to about 8 mm, about 6 mm to about 7 mm, about 6 mm to about 8 mm, or about 7 mm to about 8 mm. In some embodiments, the pupil diameter after administration is about 2 mm, about 3 mm, about 4 mm, about 5 mm, about 6 mm, about 7 mm, or about 8 mm. In some embodiments, the pupil diameter after administration is at least about 2 mm, about 3 mm, about 4 mm, about 5 mm, about 6 mm, or about 7 mm. In some embodiments, the pupil diameter after administration is at most about 3 mm, about 4 mm, about 5 mm, about 6 mm, about 7 mm, or about 8 mm.

[0027] The measurement may be the area under the curve (AUC) of the pupil diameter over 12 hours. The measurement may be the area under the curve (AUC) of the pupil diameter over 24 hours. The measurement may be the AUC of the pupil diameter from an initial starting point to infinity.

[0028] In some embodiments, the methods described herein comprise measuring the pupil diameter of an eye. In some embodiments, the measuring is performed using pupillometry. In some embodiments, pupillometry is conducted manually. In some embodiments, pupillometry isperformed with a specialized devices suitable for assessing static and / or dynamic pupil behavior. In some embodiments, the measurements are performed with a pupillometer. The pupillometer may be selected from a manual pupillometer, a digital pupillometer, an infrared pupillometer, an automated pupillometer, or a combination thereof. The measurements may be performed after administration of a formulation. The measurements may be performed at least 30 minutes, 60 minutes, 90 minutes, 120 minutes, 180 minutes or more after administration. The measurements may be compared to a reference measurement.

[0029] The reference measurement may be a measurement taken before administration of the formulation. The reference measurement may be a measurement taken from an eye administered a reference formulation at the same time point.

[0030] In some embodiments, the measurement is increased by at least about 10 % to about 100 %. In some embodiments, the measurement is increased by at least about 10 % to about 20 %, about 10 % to about 30 %, about 10 % to about 40 %, about 10 % to about 50 %, about 10 % to about 60 %, about 10 % to about 70 %, about 10 % to about 80 %, about 10 % to about 90 %, about 10 % to about 100 %, about 20 % to about 30 %, about 20 % to about 40 %, about 20 % to about 50 %, about 20 % to about 60 %, about 20 % to about 70 %, about 20 % to about 80 %, about 20 % to about 90 %, about 20 % to about 100 %, about 30 % to about 40 %, about 30 % to about 50 %, about 30 % to about 60 %, about 30 % to about 70 %, about 30 % to about 80 %, about 30 % to about 90 %, about 30 % to about 100 %, about 40 % to about 50 %, about 40 % to about 60 %, about 40 % to about 70 %, about 40 % to about 80 %, about 40 % to about 90 %, about 40 % to about 100 %, about 50 % to about 60 %, about 50 % to about 70 %, about 50 % to about 80 %, about 50 % to about 90 %, about 50 % to about 100 %, about 60 % to about 70 %, about 60 % to about 80 %, about 60 % to about 90 %, about 60 % to about 100 %, about 70 % to about 80 %, about 70 % to about 90 %, about 70 % to about 100 %, about 80 % to about 90 %, about 80 % to about 100 %, or about 90 % to about 100 %. In some embodiments, the measurement is increased by at least about 10 %, about 20 %, about 30 %, about 40 %, about 50 %, about 60 %, about 70 %, about 80 %, about 90 %, or about 100 %. In some embodiments, the measurement is increased by at least about 10 %, about 20 %, about 30 %, about 40 %, about 50 %, about 60 %, about 70 %, about 80 %, or about 90 %. In some embodiments, the measurement is increased by at most about 20 %, about 30 %, about 40 %, about 50 %, about 60 %, about 70 %, about 80 %, about 90 %, or about 100 %. The measurement may be a pupil diameter. The measurement may be an AUC.

[0031] In some embodiments, the measurement is increased for a time period following administration of at least about 30 minutes to about 300 minutes. In some embodiments, themeasurement is increased for a time period of at least about 30 minutes to about 60 minutes, about 30 minutes to about 90 minutes, about 30 minutes to about 120 minutes, about 30 minutes to about 180 minutes, about 30 minutes to about 210 minutes, about 30 minutes to about 240 minutes, about 30 minutes to about 300 minutes, about 60 minutes to about 90 minutes, about 60 minutes to about 120 minutes, about 60 minutes to about 180 minutes, about 60 minutes to about 210 minutes, about 60 minutes to about 240 minutes, about 60 minutes to about 300 minutes, about 90 minutes to about 120 minutes, about 90 minutes to about 180 minutes, about 90 minutes to about 210 minutes, about 90 minutes to about 240 minutes, about 90 minutes to about 300 minutes, about 120 minutes to about 180 minutes, about 120 minutes to about 210 minutes, about 120 minutes to about 240 minutes, about 120 minutes to about 300 minutes, about 180 minutes to about 210 minutes, about 180 minutes to about 240 minutes, about 180 minutes to about 300 minutes, about 210 minutes to about 240 minutes, about 210 minutes to about 300 minutes, or about 240 minutes to about 300 minutes. In some embodiments, the measurement is increased for a time period of at least about 30 minutes, about 60 minutes, about 90 minutes, about 120 minutes, about 180 minutes, about 210 minutes, about 240 minutes, or about 300 minutes. In some embodiments, the measurement is increased for a time period of at least about 30 minutes, about 60 minutes, about 90 minutes, about 120 minutes, about 180 minutes, about 210 minutes, or about 240 minutes. In some embodiments, the measurement is increased for a time period of at most about 60 minutes, about 90 minutes, about 120 minutes, about 180 minutes, about 210 minutes, about 240 minutes, or about 300 minutes.

[0032] In some embodiments, the measurement is increased after administration for a time period of at least about 4 hours to about 72 hours. In some embodiments, the measurement is increased for a time period of at least about 4 hours to about 8 hours, about 4 hours to about 12 hours, about 4 hours to about 18 hours, about 4 hours to about 24 hours, about 4 hours to about 32 hours, about 4 hours to about 40 hours, about 4 hours to about 48 hours, about 4 hours to about 56 hours, about 4 hours to about 64 hours, about 4 hours to about 72 hours, about 8 hours to about 12 hours, about 8 hours to about 18 hours, about 8 hours to about 24 hours, about 8 hours to about 32 hours, about 8 hours to about 40 hours, about 8 hours to about 48 hours, about 8 hours to about 56 hours, about 8 hours to about 64 hours, about 8 hours to about 72 hours, about 12 hours to about 18 hours, about 12 hours to about 24 hours, about 12 hours to about 32 hours, about 12 hours to about 40 hours, about 12 hours to about 48 hours, about 12 hours to about 56 hours, about 12 hours to about 64 hours, about 12 hours to about 72 hours, about 18 hours to about 24 hours, about 18 hours to about 32 hours, about 18 hours to about 40 hours, about 18 hours to about 48 hours, about 18 hours to about 56 hours, about 18 hours to about 64hours, about 18 hours to about 72 hours, about 24 hours to about 32 hours, about 24 hours to about 40 hours, about 24 hours to about 48 hours, about 24 hours to about 56 hours, about 24 hours to about 64 hours, about 24 hours to about 72 hours, about 32 hours to about 40 hours, about 32 hours to about 48 hours, about 32 hours to about 56 hours, about 32 hours to about 64 hours, about 32 hours to about 72 hours, about 40 hours to about 48 hours, about 40 hours to about 56 hours, about 40 hours to about 64 hours, about 40 hours to about 72 hours, about 48 hours to about 56 hours, about 48 hours to about 64 hours, about 48 hours to about 72 hours, about 56 hours to about 64 hours, about 56 hours to about 72 hours, or about 64 hours to about 72 hours. In some embodiments, the measurement is increased for a time period of at least about 4 hours, about 8 hours, about 12 hours, about 18 hours, about 24 hours, about 32 hours, about 40 hours, about 48 hours, about 56 hours, about 64 hours, or about 72 hours. In some embodiments, the measurement is increased for a time period of at least about 4 hours, about 8 hours, about 12 hours, about 18 hours, about 24 hours, about 32 hours, about 40 hours, about 48 hours, about 56 hours, or about 64 hours. In some embodiments, the measurement is increased for a time period of at most about 8 hours, about 12 hours, about 18 hours, about 24 hours, about 32 hours, about 40 hours, about 48 hours, about 56 hours, about 64 hours, or about 72 hours.

[0033] In some embodiments, the eye is a mammalian eye. In some embodiments, the eye is an avian eye. In some embodiments, the eye is a human eye. In some embodiments, the eye is a rabbit eye. In some embodiments, the eye is an adult human eye. In some embodiments, the eye is a pediatric human eye.B. Methods of identifying a formulation

[0034] Also described herein are methods for identifying a formulation of an ophthalmic composition suitable for treating myopia or premyopia. In some embodiments, the method comprises administering a formulation that increases a measurement for a time period. In some embodiments, the method comprises administering a formulation to an eye of the subject; and measuring the diameter of the pupil of the eye before administration and at a timepoint at least 60 minutes after administration. In some embodiments, the measurement comprises the diameter of the pupil. In some embodiments, the time period comprises more than 60 minutes. In some embodiments, the pupil measurement is measured as described herein.

[0035] In some embodiments, the methods described herein comprise comparing the formulation to a reference formulation. In some embodiments, the reference formulation comprises the same concentration of atropine as the formulation. The measurements may be performed after administration of the formulation and / or the reference formulation. Themeasurements may be performed at least 30 minutes, 60 minutes, 90 minutes, 120 minutes, 180 minutes or more after administration. The measurements may be compared to a reference measurement. The reference measurement may be a measurement taken before administration of the formulation and / or the reference formulation. The reference measurement may be a measurement taken from an eye administered a reference formulation at the same time point.

[0036] In some embodiments, the formulation is identified as suitable for treatment of myopia or premyopia when the formulation increases the diameter of the pupil of the eye by at least 10% for a period of 60 minutes or more. The measurements may be performed at least 30 minutes, 60 minutes, 90 minutes, 120 minutes, 180 minutes or more after administration. The measurements may be compared to a reference measurement. In some embodiments, the measurement is increased by at least about 10 %, about 20 %, about 30 %, about 40 %, about 50 %, about 60 %, about 70 %, about 80 %, about 90 %, or about 100 %.

[0037] In some embodiments, the method comprises calculating the area under the curve (AUC) of the pupil diameter for 12 hours for the formulation and the reference formulation. In some embodiments, the method comprises calculating the area under the curve (AUC) of the pupil diameter for 24 hours for the formulation and the reference formulation. In some embodiments, the formulation is identified as suitable for treatment of myopia or premyopia when the AUC of the formulation is at least 50% of the reference formulation for a period of 60 minutes or more.

[0038] Also described herein are methods of identifying a suitable formulation for treatment of myopia or premyopia, wherein the formulation does not comprise D2O. In some embodiments, the reference formulation comprises D2O. In some embodiments, the formulation is identified as suitable for treatment of myopia or premyopia when the AUC of the formulation is at least 50% of the reference formulation. In some embodiments, the formulation is identified as suitable for treatment of myopia or premyopia when the AUC of the formulation is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the AUC of the reference formulation.C. Methods of administration

[0039] The methods described herein involve the administration of a formulation of an ophthalmic composition to an eye of a subject in need thereof. In some embodiments, the formulation of the ophthalmic composition is administered to one eye of the subject. In some embodiments, the formulation of the ophthalmic composition is administered to both eyes of the subject. The formulation of the ophthalmic composition may be in the form of an ophthalmic aqueous formulation, ophthalmic gel formulation an ophthalmic suspension formulation, anophthalmic ointment formulation, an ophthalmic emulsion formulation, an ophthalmic sustained-release formulation, an ophthalmic contact lens solution formulation, and the like.

[0040] In some embodiments, the ophthalmic aqueous formulations described herein are packaged in eye drop bottles and administered as drops. For example, a single administration (e.g., a single dose) of an ophthalmic aqueous formulation includes a single drop, two drops, three drops or more into an eye of the subject. In some embodiments, the ophthalmic gel formulations described herein are packaged in eye drop bottles and administered as drops. In some embodiments, the eye drop bottle is a multidrop bottle. For example, a single administration (i.e. a single dose) of an ophthalmic gel includes a single drop, two drops, three drops or more into an eye of the subject. In some embodiments, the ophthalmic ointment formulations described herein are packaged in tubes or other squeezable containers with a dispensing nozzle through which strips of the ointment are delivered. For example, a single administration (e.g., a single dose) of an ophthalmic ointment includes a single strip, or multiple strips into the eyes of the patient. In some embodiments, the formulation is packaged in a single dose container. In some embodiments, one dose of the ophthalmic aqueous formulation described herein comprises one drop of the ophthalmic aqueous composition from the eye drop bottle. In some embodiments, one dose of the ophthalmic gel described herein is one drop of the gel composition from the eye drop bottle. In some embodiments, one dose of the ophthalmic ointment is one strip of the ointment composition dispensed through the nozzle of a dispersing tube. In some embodiments, the ophthalmic composition is not formulated as an injectable formulation.

[0041] In some embodiments, the ophthalmic composition is formulated as an ophthalmic solution for treatment of premyopia, myopia, progression of myopia, or slowing progression of myopia.

[0042] In some embodiments, the ophthalmic aqueous formulations are administered as follows: the lower lid of the eye to be administered was pulled down and a predetermined amount of the aqueous formulation (e.g. 1-3 drops) is applied to the inside of the eyelid. The ophthalmic tip of the dispensing mechanism does not touch any surface to avoid contamination and / or injury.

[0043] In some embodiments, the ophthalmic gel formulations are administered as follows: the lower lid of the eye to be administered was pulled down and a predetermined amount of gel (e.g., 1-3 drops) is applied to the inside of the eyelid. The ophthalmic tip of the dispensing mechanism does not touch any surface to avoid contamination and / or injury.

[0044] In some embodiments, the ophthalmic ointment formulations are administered as follows: the lower lid of the eye to be administered was pulled down and a small amount of ointment (approximately 0.25 inches) was applied to the inside of the eyelid. The ophthalmic tip of the dispensing mechanism does not touch any surface to avoid contamination and / or injury.

[0045] In some embodiments, administration of the ophthalmic composition comprises adherence to a dosing regimen followed by the subject. In some embodiments, one or more doses are administered at predetermined time intervals over an extended period of time. In some embodiments, the one or more doses of the ophthalmic composition is / are administered once every day. In some embodiments, the one or more doses of the ophthalmic composition is / are administered once every other day. In some embodiments, the one or more doses of the ophthalmic composition is / are administered over 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12-15 years.

[0046] In some embodiments, the number of times a composition is administered to an individual in need thereof depends on the discretion of a medical professional, the disorder, the severity of the disorder, and / or the individual’s response to the formulation. In some embodiments, a composition disclosed herein is administered once to an individual in need thereof with a mild acute condition. In some embodiments, a composition disclosed herein is administered more than once to a subject in need thereof, wherein the subject suffers from a moderate or severe acute condition. In some embodiments, the moderate or severe acute condition shows little or no improvement with or without treatment. In some embodiments, the ophthalmic composition is administered chronically (e.g., throughout the duration of the life of the subject) in order to ameliorate or otherwise control or limit the symptoms suffered by the subject.

[0047] In some embodiments, the ophthalmic composition is continuously. In some embodiments, the dose of ophthalmic composition being administered is temporarily reduced or temporarily suspended for a time period (e.g., a “drug holiday”). In some embodiments, the time period is from about 2 days to about 1 year. In some embodiments, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. In some embodiments, the initial dose of the ophthalmic composition being administered is reduced during the time period by from about 10% to about 100%. In some embodiments, the initial dose of the ophthalmic composition being administered is reduced during the time period by about 10%, about 15%, about 20%, about 25%, about 30%, about35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

[0048] In some embodiments, the ophthalmic composition is administered as a maintenance treatment (e.g., improvement in the condition suffered by the subject has occurred). In some embodiments, the administration of the ophthalmic composition as a maintenance treatment comprises one or more modifications to the frequency of administration, the dosage administered, or a combination thereof. In some embodiments, the one or more modifications is informed, at least in part, by one or more of the disease or condition suffered by the subject, severity of symptoms suffered by the subject, recurrence of existing symptoms suffered by the subject, occurrence of new symptoms suffered by the subject, the active ophthalmic agent being administered, the route of administration, the target area being treated, the subject or host being treated, concurrent administration of one or more additional ophthalmic agent(s), or a combination thereof. In some embodiments, the one or more modifications result in a dosing regimen sufficient to maintain and / or improve a disease state in the subject. In some embodiments, the frequency of administration is on an intermittent basis. In some embodiments, the frequency of administration is on a long-term basis.

[0049] In some embodiments, the ophthalmic composition is administered as single dose or as two or more doses administered simultaneously or consecutively over the course of an appropriate interval.

[0050] In some embodiments, the initial administration comprises one or more doses of a first ophthalmic agent and the subsequent or concurrent administration of a different formulation or ophthalmic agent.II. FORMULATIONS

[0051] The methods described herein comprise formulations for ophthalmic compositions. In some embodiments, the ophthalmic compositions comprise atropine. In some embodiments, the ophthalmic compositions comprise D2O. In some embodiments, the ophthalmic compositions comprise H2O. In some embodiments, the ophthalmic compositions do not comprise D2O.

[0052] In some embodiments, the atropine is present in the ophthalmic composition at a concentration of one of: from about 0.001 wt% to about 0.40 wt%, from about 0.001 wt% to about 0.30 wt%, from about 0.001 wt% to about 0.20 wt%, from about 0.001 wt% to about 0.10 wt%, from about 0.001 wt% to about 0.09 wt%, from about 0.001 wt% to about 0.08 wt%, from about 0.001 wt% to about 0.07 wt%, from about 0.001 wt% to about 0.06 wt%, from about0.001 wt% to about 0.05 wt%, from about 0.001 wt% to about 0.04 wt%, from about 0.001 wt% to about 0.03 wt%, from about 0.001 wt% to about 0.025 wt%, from about 0.001 wt% to about 0.02 wt%, from about 0.001 wt% to about 0.01 wt%, from about 0.001 wt% to about 0.008 wt%, or from about 0.001 wt% to about 0.005 wt%. In some embodiments, the atropine is at a concentration of 0.001 wt% to 0.1 wt%. In some embodiments, the ophthalmic composition further comprises an osmolarity adjusting agent. In some embodiments, the osmolarity adjusting agent is sodium chloride. In some embodiments, the sodium chloride is present in the ophthalmic composition at a concentration of one of: from about 0.01 wt% to about lwt%, from about 0.05 wt% to about 1.5 wt%, from about 0.075 wt% to about 2wt%, or from about 0.1 wt% to about 3wt%. In some embodiments, the ophthalmic composition further comprises a buffering agent. In some embodiments, the buffering agent comprises citrate.

[0053] In some embodiments of an ophthalmic composition described herein, the ophthalmic composition comprises atropine or its pharmaceutically acceptable salt. In some embodiments, atropine or its pharmaceutically acceptable salt is present in the ophthalmic composition at a concentration of one of: from about 0.001 wt% to about 0.40 wt%, from about 0.001 wt% to about 0.30 wt%, from about 0.001 wt% to about 0.20 wt%, from about 0.001 wt% to about 0.10 wt%, from about 0.001 wt% to about 0.09 wt%, from about 0.001 wt% to about 0.08 wt%, from about 0.001 wt% to about 0.07 wt%, from about 0.001 wt% to about 0.06 wt%, from about 0.001 wt% to about 0.05 wt%, from about 0.001 wt% to about 0.04 wt%, from about 0.001 wt% to about 0.03 wt%, from about 0.001 wt% to about 0.025 wt%, from about 0.001 wt% to about 0.02 wt%, from about 0.001 wt% to about 0.01 wt%, from about 0.001 wt% to about 0.008 wt%, or from about 0.001 wt% to about 0.005 wt%. In some embodiments of an ophthalmic composition described herein, the atropine or its pharmaceutically acceptable salt is present in the ophthalmic composition at a concentration from about 0.001 wt% to about 0. 10 wt%.

[0054] In some embodiments of an ophthalmic composition described herein, the composition comprises a muscarinic antagonist. In some embodiments, the muscarinic antagonist is present in the ophthalmic composition at a concentration of one of: from about 0.001 mg / g to about 0.40 mg / g, from about 0.001 mg / g to about 0.30 mg / g, from about 0.001 mg / g to about 0.20 mg / g, from about 0.001 mg / g to about 0.10 mg / g, from about 0.001 mg / g to about 0.09 mg / g, from about 0.01 mg / g to about 0.40 mg / g, from about 0.01 mg / g to about 0.30 mg / g, from about 0.01 mg / g to about 0.20 mg / g, from about 0.01 mg / g to about 0.10 mg / g, from about 0.01 mg / g to about 0.09 mg / g, from about 0.01 mg / g to about 0.08 mg / g, from about 0.01 mg / g to about 0.07 mg / g, from about 0.01 mg / g to about 0.06 mg / g, from about 0.01 mg / g toabout 0.05 mg / g, from about 0.01 mg / g to about 0.04 mg / g, from about 0.01 mg / g to about 0.03 mg / g, from about 0.01 mg / g to about 0.025 mg / g, from about 0.01 mg / g to about 0.02 mg / g, from about 0.01 mg / g to about 0.1 mg / g, from about 0.01 mg / g to about 0.25 mg / g, from about 0.01 mg / g to about 0.5 mg / g, from about 0.01 mg / g to about 0.75 mg / g, from about 0.01 mg / g to about 1.0 mg / g. In some embodiments of an ophthalmic composition described herein, the muscarinic antagonist is present in the ophthalmic composition at a concentration from about 0.01 mg / g to about 0.5 mg / g.

[0055] In some embodiments of an ophthalmic composition described herein, the atropine or its pharmaceutically acceptable salt is present in the ophthalmic composition at a concentration of one of: from about 0.001 mg / g to about 0.40 mg / g, from about 0.001 mg / g to about 0.30 mg / g, from about 0.001 mg / g to about 0.20 mg / g, from about 0.001 mg / g to about 0.10 mg / g, from about 0.001 mg / g to about 0.09 mg / g, from about 0.01 mg / g to about 0.40 mg / g, from about 0.01 mg / g to about 0.30 mg / g, from about 0.01 mg / g to about 0.20 mg / g, from about 0.01 mg / g to about 0.10 mg / g, from about 0.01 mg / g to about 0.09 mg / g, from about 0.01 mg / g to about 0.08 mg / g, from about 0.01 mg / g to about 0.07 mg / g, from about 0.01 mg / g to about 0.06 mg / g, from about 0.01 mg / g to about 0.05 mg / g, from about 0.01 mg / g to about 0.04 mg / g, from about 0.01 mg / g to about 0.03 mg / g, from about 0.01 mg / g to about 0.025 mg / g, from about 0.01 mg / g to about 0.02 mg / g, from about 0.01 mg / g to about 0.1 mg / g, from about 0.01 mg / g to about 0.25 mg / g, from about 0.01 mg / g to about 0.5 mg / g, from about 0.01 mg / g to about 0.75 mg / g, from about 0.01 mg / g to about 1.0 mg / g. In some embodiments of an ophthalmic composition described herein, the atropine or its pharmaceutically acceptable salt is present in the ophthalmic composition at a concentration from about 0.01 mg / g to about 0.5 mg / g.

[0056] In some embodiments of an ophthalmic composition described herein, the muscarinic antagonist is present in the ophthalmic composition in an amount of one of: from about 0.0001 mg to about 0.040 mg, from about 0.0001 mg to about 0.030 mg, from about 0.0001 mg to about 0.020 mg, from about 0.0001 mg to about 0.010 mg, from about 0.0001 mg to about 0.009 mg, from about 0.001 mg to about 0.040 mg, from about 0.001 mg to about 0.030 mg, from about 0.001 mg to about 0.020 mg, from about 0.001 mg to about 0.010 mg, from about 0.001 mg to about 0.009 mg, from about 0.001 mg to about 0.008 mg, from about 0.001 mg to about 0.007 mg, from about 0.001 mg to about 0.006 mg, from about 0.001 mg to about 0.005 mg, from about 0.001 mg to about 0.004 mg, from about 0.001 mg to about 0.003 mg, from about 0.001 mg to about 0.0025 mg, from about 0.001 mg to about 0.002 mg, from about 0.001 mg to about 0.01 mg, from about 0.001 mg to about 0.025 mg, from about 0.001 mg to about 0.05 mg, from about 0.001 mg to about 0.075 mg, from about 0.001 mg to about Img. In some embodiments ofan ophthalmic composition described herein, the muscarinic antagonist is present in the ophthalmic composition in an amount from about 0.0003 mg to about 0.025 mg or 0.001 mg to about 0.05 mg.

[0057] In some embodiments of an ophthalmic composition described herein, the atropine or its pharmaceutically acceptable salt is present in the ophthalmic composition in an amount of one of: from about 0.0001 mg to about 0.040 mg, from about 0.0001 mg to about 0.030 mg, from about 0.0001 mg to about 0.020 mg, from about 0.0001 mg to about 0.010 mg, from about 0.0001 mg to about 0.009 mg, from about 0.001 mg to about 0.040 mg, from about 0.001 mg to about 0.030 mg, from about 0.001 mg to about 0.020 mg, from about 0.001 mg to about 0.010 mg, from about 0.001 mg to about 0.009 mg, from about 0.001 mg to about 0.008 mg, from about 0.001 mg to about 0.007 mg, from about 0.001 mg to about 0.006 mg, from about 0.001 mg to about 0.005 mg, from about 0.001 mg to about 0.004 mg, from about 0.001 mg to about 0.003 mg, from about 0.001 mg to about 0.0025 mg, from about 0.001 mg to about 0.002 mg, from about 0.001 mg to about 0.01 mg, from about 0.001 mg to about 0.025 mg, from about 0.001 mg to about 0.05 mg, from about 0.001 mg to about 0.075 mg, from about 0.001 mg to about Img. In some embodiments of an ophthalmic composition described herein, the atropine or its pharmaceutically acceptable salt is present in the ophthalmic composition in an amount from about 0.0003 mg to about 0.025 mg or 0.001 mg to about 0.05 mg.

[0058] In some aspects, described herein are formulations or solutions comprising a muscarinic antagonist (e.g., atropine) formulated in deuterated water (e.g., D2O). In some aspects, formulations, or solutions comprising a muscarinic antagonist (e.g., atropine) formulated in deuterated water are stable at various temperatures, humidities, pD ranges (e.g., an acidic pD), while having a potency of at least 80% relative to the ophthalmic agent. In additional aspects, ophthalmic formulations or solutions comprising a muscarinic antagonist (e.g., atropine) formulated in deuterated water has a lowered buffering capacity. In some instances, the lowered buffering capacity of the ophthalmic formulations or solutions when administered into the eye allows the ophthalmic formulation or solution to reach physiological pH at a faster rate than compared to an equivalent ophthalmic formulation or solution formulated in H2O.

[0059] Described herein, in some embodiments, is an ophthalmic composition comprising ethylenediaminetetraacetic acid (EDTA). In some embodiments, EDTA is present in the composition at about 0.001%, 0.005%, 0.010%, 0.015%, 0.020%, 0.025%, 0.030%, 0.035%, 0.040%, 0.045%, 0.050%, 0.055%, 0.060%, 0.065%, 0.070%, 0.075%, 0.080%, 0.085%, 0.090%, 0.095%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.5%, 2.0%, 2.5%, or 3.0%. In some embodiments, EDTA is present in the composition from about 0.01% toabout 0.05%, from about 0.01% to about 0.04%, from about 0.01% to about 0.03%, from about 0.01% to about 0.025%, from about 0.01% to about 0.02%, from about 0.001% to about 0.01%, from about 0.001% to about 0.008%, or from about 0.001% to about 0.005%. In some cases, the percentage is a weight percentage (e.g., wt %).

[0060] In some embodiments, the composition has a pH of from about 4 to about 8, about 4.2 to about 7.9, about 4.5 to about 7.8, about 5 to about 7.5, about 4.9 to about 6.1, about 5to about 6, about 5.2 to about 6.1, about 5.5 to about 7, or about 5.5 to about 5.6, for example when measured at about 25 °C. In some embodiments, the composition has a pH of from about 4 to about 8, about 4.2 to about 7.9, about 4.5 to about 7.8, about 5 to about 7.5, about 4.9 to about 6.1, about 5to about 6.0, about 5.2 to about 6.1, about 5.5 to about 7, or about 5.5 and 5.6, for example when measured at about 40 °C. In some embodiments, the composition has a pH of about 8. In some embodiments, the composition has a pH of about 7.9. In some embodiments, the composition has a pH of about 7.8. In some embodiments, the composition has a pH of about 7.7. In some embodiments, the composition has a pH of about 7.6. In some embodiments, the composition has a pH of less than about 7.5. In some embodiments, the composition has a pH of less than about 7.4. In some embodiments, the composition has a pH of less than about 7.3. In some embodiments, the composition has a pH of less than about 7.2. In some embodiments, the composition has a pH of less than about 7. 1. In some embodiments, the composition has a pH of less than about 7. In some embodiments, the composition has a pH of less than about 6.9. In some embodiments, the composition has a pH of less than about 6.8. In some embodiments, the composition has a pH of less than about 6.7. In some embodiments, the composition has a pH of less than about 6.6. In some embodiments, the composition has a pH of less than about 6.5. In some embodiments, the composition has a pH of less than about 6.4. In some embodiments, the composition has a pH of less than about 6.3. In some embodiments, the composition has a pH of less than about 6.2. In some embodiments, the composition has a pH of less than about 6.1. In some embodiments, the composition has a pH of less than about 6. In some embodiments, the composition has a pH of less than about 5.9. In some embodiments, the composition has a pH of less than about 5.8. In some embodiments, the composition has a pH of less than about 5.7. In some embodiments, the composition has a pH of less than about 5.6. In some embodiments, the composition has a pH of less than about 5.5. In some embodiments, the composition has a pH of less than about 5.4. In some embodiments, the composition has a pH of less than about 5.3. In some embodiments, the composition has a pH of less than about 5.2. In some embodiments, the composition has a pH of less than about 5. 1. In some embodiments, the composition has a pH of less than about 5. In some embodiments, the composition has a pH of less than about 4.9. Insome embodiments, the composition has a pH of less than about 4.8. In some embodiments, the composition has a pH of less than about 4.7. In some embodiments, the composition has a pH of less than about 4.6. In some embodiments, the composition has a pH of less than about 4.5. In some embodiments, the composition has a pH of less than about 4.4. In some embodiments, the composition has a pH of less than about 4.3. In some embodiments, the composition has a pH of less than about 4.2. In some embodiments, the composition has a pH of less than about 4.1. In some embodiments, the composition has a pH of less than about 4.

[0061] In some embodiments, the composition has a pD of from about 4 to about 8, about 4.2 to about 7.9, about 4.5 to about 7.8, about 5 to about 7.5, about 5.5 to about 7, about 5.3 to about 6.5, about 5.4 to about 6.4, about 5.6 to about 6.5, or about 5.9 and about6.0, for example when measured at about 25 °C. In some embodiments, the composition has a pD of from about 4 to about 8, about 4.2 to about 7.9, about 4.5 to about 7.8, about 5 to about 7.5, about 5.5 to about 7, about 5.3 to about 6.5, about 5.4 to about 6.4, about 5.6 to about 6.5, or about 5.9 and about 6.0, for example when measured at about 40 °C. In some embodiments, the composition has a pD of about 8. In some embodiments, the composition has a pD of about 7.9. In some embodiments, the composition has a pD of about 7.8. In some embodiments, the composition has a pD of about 7.7. In some embodiments, the composition has a pD of about 7.6. In some embodiments, the composition has a pD of less than about 7.5. In some embodiments, the composition has a pD of less than about 7.4. In some embodiments, the composition has a pD of less than about 7.3. In some embodiments, the composition has a pD of less than about 7.2. In some embodiments, the composition has a pD of less than about 7.1. In some embodiments, the composition has a pD of less than about 7. In some embodiments, the composition has a pD of less than about 6.9. In some embodiments, the composition has a pD of less than about 6.8. In some embodiments, the composition has a pD of less than about 6.7. In some embodiments, the composition has a pD of less than about 6.6. In some embodiments, the composition has a pD of less than about 6.5. In some embodiments, the composition has a pD of less than about 6.4. In some embodiments, the composition has a pD of less than about 6.3. In some embodiments, the composition has a pD of less than about 6.2. In some embodiments, the composition has a pD of less than about 6.1. In some embodiments, the composition has a pD of less than about 6. In some embodiments, the composition has a pD of less than about 5.9. In some embodiments, the composition has a pD of less than about 5.8. In some embodiments, the composition has a pD of less than about 5.7. In some embodiments, the composition has a pD of less than about 5.6. In some embodiments, the composition has a pD of less than about 5.5. In some embodiments, the composition has a pD of less than about 5.4. In some embodiments, the composition has a pD ofless than about 5.3. In some embodiments, the composition has a pD of less than about 5.2. In some embodiments, the composition has a pD of less than about 5.1. In some embodiments, the composition has a pD of less than about 5. In some embodiments, the composition has a pD of less than about 4.9. In some embodiments, the composition has a pD of less than about 4.8. In some embodiments, the composition has a pD of less than about 4.7. In some embodiments, the composition has a pD of less than about 4.6. In some embodiments, the composition has a pD of less than about 4.5. In some embodiments, the composition has a pD of less than about 4.4. In some embodiments, the composition has a pD of less than about 4.3. In some embodiments, the composition has a pD of less than about 4.2. In some embodiments, the composition has a pD of less than about 4.1. In some embodiments, the composition has a pD of less than about 4.

[0062] In some instances, the ophthalmic composition further comprises an osmolarity adjusting agent. In some cases, the osmolarity adjusting agent is sodium chloride.

[0063] In some instances, the ophthalmic composition further comprises a preservative. In some cases, the preservative is selected from benzalkonium chloride, cetrimonium, sodium perborate, stabilized oxychloro complex, SofZia, polyquatemium-1, chlorobutanol, edetate disodium, polyhexamethylene biguanide, or combinations thereof.

[0064] In some instances, the ophthalmic composition further comprises a buffer agent. In some cases, the buffer agent is selected from borates, borate-polyol complexes, phosphate buffering agents, citrate buffering agents, acetate buffering agents, carbonate buffering agents, organic buffering agents, amino acid buffering agents, or combinations thereof.

[0065] In some instances, the ophthalmic composition further comprises a tonicity adjusting agent. In some cases, the tonicity adjusting agent is selected from sodium chloride, sodium nitrate, sodium sulfate, sodium bisulfate, potassium chloride, calcium chloride, magnesium chloride, zinc chloride, potassium acetate, sodium acetate, sodium bicarbonate, sodium carbonate, sodium thiosulfate, magnesium sulfate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dextrose, mannitol, sorbitol, dextrose, sucrose, urea, propylene glycol, glycerin, or a combination thereof.

[0066] In some instances, the ophthalmic composition further comprises a penetration agent. In some instances, the penetration agent is benzalkonium chloride.III. DEFINITIONS

[0067] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. Insome cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.

[0068] Throughout this application, various embodiments may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0069] As used in the specification and claims, the singular forms “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a sample” includes a plurality of samples, including mixtures thereof.

[0070] The terms “determining,” “measuring,” “evaluating,” “assessing,” “assaying,” and “analyzing” are often used interchangeably herein to refer to forms of measurement. The terms include determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing can be relative or absolute. “Detecting the presence of’ can include determining the amount of something present in addition to determining whether it is present or absent depending on the context.

[0071] The terms “subject,” “individual,” or “patient” are often used interchangeably herein. A “subject” can be a biological entity containing expressed genetic materials. The biological entity can be a plant, animal, or microorganism, including, for example, bacteria, viruses, fungi, and protozoa. The subject can be tissues, cells and their progeny of a biological entity obtained in vivo or cultured in vitro. The subject can be a mammal. The mammal can be a human. The subject may be diagnosed or suspected of being at high risk for a disease. In some cases, the subject is not necessarily diagnosed or suspected of being at high risk for the disease.

[0072] The term “in vivo " is used to describe an event that takes place in a subject’s body.

[0073] The term ex vivo is used to describe an event that takes place outside of a subject s body. An ex vivo assay is not performed on a subject. Rather, it is performed upon a sample separate from a subject. An example of an ex vivo assay performed on a sample is an “in vitro” assay.

[0074] The term “in vitro” is used to describe an event that takes places contained in a container for holding laboratory reagent such that it is separated from the biological source from which the material is obtained. In vitro assays can encompass cell-based assays in which living or dead cells are employed. In vitro assays can also encompass a cell-free assay in which no intact cells are employed.

[0075] As used herein, the term “about” a number refers to that number plus or minus 10% of that number. The term “about” a range refers to that range minus 10% of its lowest value and plus 10% of its greatest value.

[0076] As used herein, the terms “treatment” or “treating” are used in reference to a pharmaceutical or other intervention regimen for obtaining beneficial or desired results in the recipient. Beneficial or desired results include but are not limited to a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit may refer to eradication or amelioration of symptoms or of an underlying disorder being treated. Also, a therapeutic benefit can be achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. A prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof. For prophylactic benefit, a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease may undergo treatment, even though a diagnosis of this disease may not have been made.

[0077] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.IV. EXAMPLES

[0078] The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention.Example 1: In vivo Ocular Response Testing in Dutch Belted Rabbits Using Atropine Formulations

[0079] A study was conducted to investigate the ocular response to two atropine formulations in Dutch Belted (DB) rabbits. The experimental protocol is depicted in FIG. 3. First, 60 DB rabbits (30 males and 30 females) obtained from Western Oregon Rabbit Companywere screened for atropinesterase levels. Plasma samples from each rabbit were incubated with 150pM atropine stock solutions for two hours, then analyzed for tropic acid concentrations. Based on this screening, 20 rabbits with tropic acid concentrations ranging from 39.1 ng / mL to 79 ng / mL, indicating low atropinesterase levels, were selected for further testing. A one-week washout period was observed post screening. Following the washout period, the selected 20 rabbits (10 males and 10 females) underwent functional testing to validate pupil dilation of each rabbit via administration of a 0.02% Atropine Sulfate Ophthalmic Solution, obtained by diluting a 1% solution from Amneal Pharmaceuticals with Bausch & Lomb Advanced Eyewash (containing 0.01% BAK, Borate Buffer, and NaCl at a pH of 7). A dosage of the Atropine Sulfate Ophthalmic Solution, in the amount of 35 pL, was administered to each eye in each rabbit. After dosing, evaluations were conducted, including pupillometry at nine different timepoints. Pupillometry measurements were taken within six hours post-dose using a NeurOptics VIP -400 Pupillometer. Each measurement was taken in triplicate. 12 rabbits were then selected for further study and then given a one-week washout period. Both groups were then subjected to the same pupillometry evaluations at nine timepoints. Post-evaluation, a one- week washout period was observed before the treatments for each group were crossed over. The same evaluations were repeated after the crossover. Results are depicted in FIGS. 1A-1B and 2A-2D. The Area Under the Curve (AUC) was calculated for t=0- 12 hours post-dose based on both eyes (OU) pupil diameter measurements, considering the average of the left eye (OS), and right eye (OD). The AUC for TA2, comprising 0.02 wt% Atropine, deuterated water, and 0.01% BAK, was determined to be 612.02 mm*min, whereas for TA3, comprising H2O and no BAK, the AUC was significantly larger at 1235.27 mm*min. When the results were extrapolated out to 24 hours, the AUC for TA3 was 1717.67 mm*min and 714.97 for TA2 mm*min.Example 2: Identifying a formulation suitable for treatment of premyopia or myopia

[0080] A formulation is identified as suitable for the treatment of premyopia or myopia by administering the formulation to the eye of a mammalian. The subject may be a rabbit. The test may be repeated on a plurality of rabbits.

[0081] The diameter of the pupil is measured via pupillometry at least one, two, three or more timepoints after administration. At least one timepoint is before administration, and at least one time point is 60 minutes or greater following administration. One time point may be 12 hours after administration. One time point may be 24 hours after administration.

[0082] A reference formulation may be administered. The reference formulation may comprise atropine at the same concentration as the test formulation. The reference formulationmay comprise BAK, may be formulated in D2O, and may have a pD of 7. The reference formulation may comprise BAK, be formulated in D2O, and have a pD of 7.4.

[0083] The area under the curve (AUC) of the pupil diameter for 12 hours is calculated for the formulation and the reference formulation. The area under the curve (AUC) of the pupil diameter for 24 hours is calculated for the formulation and the reference formulation. The formulation may be identified as suitable for treatment of myopia or premyopia when the AUC of the formulation is at least 50% of the AUC of the reference formulation for a period of 60 minutes or more. The formulation may be identified as suitable for treatment of myopia or premyopia when the formulation increases the diameter of the pupil of the eye by at least 10% for a period of 60 minutes or more.While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

CLAIMS1. A method of identifying a formulation suitable for treatment of premyopia or myopia, the method comprising administering the formulation that increases the diameter of the pupil of the eye by at least 10% for a period of 60 minutes or more.

2. The method of claim 1, wherein the diameter of the pupil is measured by pupillometry.

3. The method of claim 1 or 2, wherein the diameter is increased by at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90 % for a period of 60 minutes or more.

4. The method of any one of claims 1-3, wherein the diameter is increased for a period of 90 minutes or more.

5. A method of identifying a formulation suitable for treatment of premyopia or myopia, the method comprising: a. administering a formulation to an eye of the subject; and b. measuring the diameter of the pupil of the eye before administration and at a timepoint at least 60 minutes after administration.

6. The method of claim 5, wherein the diameter of the pupil is measured by pupillometry.

7. The method of claim 5 or 6, wherein the diameter of the pupil is measured at two or more timepoints after administration.

8. The method of any one of claims 5-7, wherein the diameter is increased by at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90 % for a period of 60 minutes or more.

9. The method of any one of claims 5-8, wherein the diameter is increased for a period of 90 minutes or more.

10. The method of any one of claims 1-9, wherein the eye is a mammalian eye.

11. The method of any one of claims 1-10, wherein the eye is a human eye.

12. The method of any one of claims 1-10, wherein the eye is a rabbit eye.

13. The method of any one of claims 1-10, wherein the eye is an adult human eye.

14. The method of any one of claims 1-10, wherein the eye is a pediatric human eye.

15. The method of any one of claims 1-14, wherein the pupil diameter prior to administration is from about 2 mm to about 7 mm.

16. The method of any one of claims 1-14, wherein the pupil diameter prior to administration is from about 2 mm to about 8 mm.

17. The method of any one of claims 1-16, wherein the formulation is administered to only one eye of the subject.

18. The method of any one of claims 1-16, wherein the formulation is administered to both eyes of the subject.

19. The method of any one of claims 1-18, wherein the formulation is compared to a reference formulation.

20. The method of claim 19, wherein the reference formulation comprises D2O.

21. The method of claim 19, wherein the reference formula comprises BAK.

22. The method of claim 19, wherein the reference formula has a pD of about 7 or a pH of about 6.6.

23. The method of claim 19, wherein the reference formula comprises BAK, is formulated in D2O, and has a pD of about 7.

24. The method of any one of claims 19-23, wherein the reference formulation comprises the same concentration of atropine as the formulation.

25. The method of any one of claims 19-24, wherein the formulation is identified as suitable for treatment of myopia or premyopia when the formulation increases the diameter of the pupil of the eye by at least 10% for a period of 60 minutes or more.

26. The method of any one of claims 19-25, further comprising calculating the area under the curve (AUC) of the pupil diameter for 12 hours for the formulation and the reference formulation.

27. The method of any one of claims 19-25, further comprising calculating the area under the curve (AUC) of the pupil diameter for 24 hours for the formulation and the reference formulation.

28. The method of claim 26 or 27, wherein the formulation is identified as suitable for treatment of myopia or premyopia when the AUC of the formulation is at least 50% of the reference formulation for a period of 60 minutes or more, wherein the reference formulation comprises D2O and the formulation does not comprise D2O.

29. A method of assessing potency of a water-based low -dose atropine formulation by administering a drop of said formulation to the eye of a subject and measuring the diameter of a pupil of an eye for a period of 60 minutes or more and comparing said formulation to a second reference formulation of an identical concentration of low-dose atropine also comprising D2O and BAK.

30. A method of increasing the diameter of a pupil of an eye by at least 10% for a period of 60 minutes or more, comprising administering a formulation comprising atropine, D2O, and BAK to the eye.

31. The method of claim 30, wherein the formulation comprises a pD of about 7.

32. The method of claim 30 or 31, wherein the diameter is increased relative to a reference formulation comprising atropine at the same dose, wherein the formulation does not comprise BAK.

33. The method of claim 32, wherein the reference formulation comprises a pD of less than about 4 or a pH of less than or about 3.6.

34. The method of any one of claims 30-33, wherein the diameter is increased by at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90 % for a time period of 60 minutes or more.

35. The method of any one of claims 30-34, wherein the diameter is increased for a time period of 90 minutes or more.

36. The method of any one of claims 30-35, wherein the eye is a mammalian eye.

37. The method of any one of claims 30-36, wherein the eye is a human eye.

38. The method of any one of claims 30-37, wherein the eye is an adult human eye.

39. The method of any one of claims 30-37, wherein the eye is a pediatric human eye.

40. The method of any one of claims 30-39, wherein the pupil diameter prior to administration is from about 2mm and to about 7 mm.

41. The method of any one of claims 30-39, wherein the pupil diameter prior to administration is from about 2 mm and to about 8 mm.

42. The method of any one of claims 30-41, wherein the formulation is administered to only one eye of the subject.

43. The method of any one of claims 30-41, wherein the formulation is administered to both eyes of the subject.

44. The method of any one of claims 30-43, wherein the atropine is present in the ophthalmic composition at a concentration of one of: from about 0.001 wt% to about 0.40 wt%, from about 0.001 wt% to about 0.30 wt%, from about 0.001 wt% to about 0.20 wt%, from about 0.001 wt% to about 0.10 wt%, from about 0.001 wt% to about 0.09 wt%, from about 0.001 wt% to about 0.08 wt%, from about 0.001 wt% to about 0.07 wt%, from about 0.001 wt% to about 0.06 wt%, from about 0.001 wt% to about 0.05 wt%, from about 0.001 wt% to about 0.04 wt%, from about 0.001 wt% to about 0.03 wt%, from about 0.001 wt% to about 0.025 wt%, from about 0.001 wt% to about 0.02 wt%, from about 0.001 wt% to about 0.01 wt%, from about 0.001 wt% to about 0.008 wt%, or from about 0.001 wt% to about 0.005 wt%.

45. The method of claim 44, wherein the atropine is at a concentration of 0.001 wt% to 0. 1 wt%.

46. The method of any one of claims 30-45, wherein the ophthalmic composition further comprises an osmolarity adjusting agent.

47. The method of claim 46, wherein the osmolarity adjusting agent is sodium chloride.

48. The method of claim 47, wherein the sodium chloride is present in the ophthalmic composition at a concentration of one of: from about 0.01 wt% to about 1.0 wt%, from about 0.05 wt% to about 1.5 wt%, from about 0.075 wt% to about 2.0 wt%, or from about 0.1 wt% to about 3.0 wt%.

49. The method of any one of claims 30-48, wherein the ophthalmic composition further comprises a buffering agent.

50. The method of claim 49, wherein the buffering agent comprises citrate.

51. A method of increasing the diameter of a pupil of an eye by at least 10% for a period of 60 minutes or more, comprising administering a formulation comprising atropine, and BAK to the eye, wherein the composition comprises a pD of about 7 or a pH of about 6.6.

52. The method of claim 51, wherein the composition does not comprise D2O.

53. The method of claim 51, wherein the composition comprises D2O.

54. The method of any one of claim 51 or 52, wherein the diameter is increased relative to a reference formulation comprising atropine at the same dose, wherein the reference formulation does not comprise BAK.

55. The method of claim 54, wherein the reference formulation comprises a pD of less than about 4 or a pH of less than about 3.6.

56. The method of any one of claims 51-55, wherein the diameter is increased by at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90 % for a time period of 60 minutes or more.

57. The method of any one of claims 51-56, wherein the diameter is increased for a time period of 90 minutes or more.

58. The method of any one of claims 51-57, wherein the eye is a mammalian eye.

59. The method of any one of claims 51-58, wherein the eye is a human eye.

60. The method of any one of claims 51-59, wherein the eye is an adult human eye.

61. The method of any one of claims 51-59, wherein the eye is a pediatric human eye.

62. The method of any one of claims 51-61, wherein the pupil diameter prior to administration is between 2mm and 7 mm.

63. The method of any one of claims 51-61, wherein the pupil diameter prior to administration is between 2 mm and 8 mm.

64. The method of any one of claims 51-63, wherein the formulation is administered to only one eye of the subject.

65. The method of any one of claims 51-63, wherein the formulation is administered to both eyes of the subject.

66. The method of any one of claims 51-65, wherein the atropine is present in the ophthalmic composition at a concentration of one of: from about 0.001 wt% to about 0.40 wt%, from about 0.001 wt% to about 0.30 wt%, from about 0.001 wt% to about 0.20 wt%, from about 0.001 wt% to about 0. 10 wt%, from about 0.001 wt% to about 0.09 wt%, from about 0.001 wt% to about 0.08 wt%, from about 0.001 wt% to about 0.07 wt%, from about 0.001 wt% to about 0.06 wt%, from about 0.001 wt% to about 0.05 wt%, from about 0.001 wt% to about 0.04 wt%, from about 0.001 wt% to about 0.03 wt%, from about 0.001 wt% to about 0.025 wt%, from about 0.001 wt% to about 0.02 wt%, from about 0.001 wt% to about 0.01 wt%, from about 0.001 wt% to about 0.008 wt%, or from about 0.001 wt% to about 0.005 wt%.

67. The method of claim 66, wherein the atropine is at a concentration of 0.001 wt% to 0. 1 wt%.

68. The method of any one of claims 51-67, wherein the ophthalmic composition further comprises an osmolarity adjusting agent.

69. The method of claim 68, wherein the osmolarity adjusting agent is sodium chloride.

70. The method of claim 69, wherein the sodium chloride is present in the ophthalmic composition at a concentration of one of: from about 0.01 wt% to about 1.0 wt%, from about 0.05 wt% to about 1.5 wt%, from about 0.075 wt% to about 2.0 wt%, or from about 0.1 wt% to about 3.0 wt%.

71. The method of any one of claims 51-70, wherein the ophthalmic composition further comprises a buffering agent.

72. The method of claim 71, wherein the buffering agent comprises citrate.

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