Liquid composition

Abstract

An ophthalmically-acceptable aqueous liquid composition is provided comprising an amine buffer and a muscarinic acetylcholine receptor antagonist which comprises a hydrolysable bond wherein the muscar

Description

FIELD OF THE INVENTION

[001] The field of the invention relates to an ophthalmically-acceptable liquid composition comprising an amine buffer and a muscarinic acetylcholine receptor antagonist complexed with a macrocyclic host molecule such as cyclodextrin or cucurbituril. The liquid composition may be stored in a heat-sterilized container. Methods of manufacturing such containers are also described. BACKGROUND

[002] Pharmaceutical agents containing hydrolysable bonds can be unstable in aqueous liquid formulations, particularly when exposed to heat, resulting in decomposition to inactive, or toxic by-products. Liquid formulations containing such unstable agents can have a short shelf-life and thus often need to be made up at the point of care, or when prescribed, from a lyophilized powder.

[003] Atropine is a myopia management agent which safely and effectively slows down myopia progression. Under heated conditions or during long term storage, atropine degrades and produces by-products. In aqueous solutions, the major byproducts are dehydration product apoatropine and hydrolysis products of tropic acid and tropine. Among the degradation byproducts, apoatropine is about 20 times more toxic than atropine, which may raise the risk of drug safety or lead to reduced atropine dosage. There is a need to improve atropine’s stability in aqueous solutions, minimizing toxicity with less dehydration products and increasing efficacy of atropine.

[004] Atropine is most stable around pH 4.0. The administration of eye drops at this acidic pH is uncomfortable for patients. There is a need for a stable atropine composition with a higher pH, which would be more acceptable to patients.

[005] Pirenzepine is another antimuscarinic agent and is commonly used to treat duodenal ulcers. It is also effective for controlling myopia progression in children. Although more stable than atropine, pirenzepine is still subject to significant hydrolysis, especially upon exposure to heat.

[006] Liquid pharmaceutical formulations typically contain a phosphate-based buffer, such as phosphate buffered saline. In the present invention, amine buffers are used as these have been found to reduce the hydrolysis of muscarinic acetylcholine receptor antagonists in aqueous formulations. In addition, it has been found that the hydrolysis of muscarinic acetylcholine receptor antagonists can be further reduced by forming a complex with a macrocyclic host molecule such as a cyclodextrin or a cucurbituril. SUMMARY

[007] The present invention provides an ophthalmically-acceptable liquid composition comprising a muscarinic acetylcholine receptor antagonist complexed with a macrocyclic host molecule. The composition is preferably heat-stable.

[008] The present invention also provides a container containing a heat-stable ophthalmically-acceptable liquid composition of the invention.

[009] The present invention also provides a method of manufacturing a heat-sterilized container of the invention comprising: (a) placing an ophthalmically-acceptable solution in a container, (b) sealing the container, and (c) heat sterilizing the sealed container, wherein the solution is a liquid ophthalmically-acceptable formulation of the invention.

[010] Additional features and advantages of the present invention will be set forth in part in the description that follows, and in part will be apparent from the description, or may be learned by practice of the present invention. The objectives and other advantages of the present invention will be realized and attained by means of the elements and combinations particularly pointed out in the description and appended claims. [Oil] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are intended to provide a further explanation of the present invention, as claimed.

[012] Unless stated otherwise, all % are weight% and are based on the total weight of the composition or solution. DETAILED DESCRIPTION

[013] The present invention provides an ophthalmically-acceptable liquid composition comprising a muscarinic acetylcholine receptor antagonist complexed with a macrocyclic host molecule, which is stable when subjected to heat such as through heat sterilization. The ophthalmically-acceptable liquid composition comprises an amine buffer and a muscarinic acetylcholine receptor antagonist, wherein the muscarinic acetylcholine receptor antagonist comprises a hydrolysable bond.

[014] Typically, the liquid composition has a pH within the range of 5.0-7.5. Preferably the composition has a pH of 6.2 to 7.4, more preferably 6.3 to 7.2, more preferably 6.5 to 7.0, even more preferably 6.8 to 7.0. The pH is determined using a calibrated bench-top pH meter such as the Hanna HI 9321 benchtop pH meter or equivalent.

[015] The liquid composition is an aqueous composition which comprises an amine buffer which has an effective pH within the range 5.0-7.5. As used herein, an “amine buffer” comprises a buffering agent which contains an amine group. In one example, the amine group-containing buffering agent is the only buffering agent included in the composition. The amine group can be a primary, secondary or tertiary amine group.

[016] Suitable amine buffers or amine group-containing buffering agents include Tris(hydroxymethyl)aminomethane (Tris), N-(2-acetamido)iminodiacetic acid (ADA), bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane (Bis-Tris), 2-morpholinoethanesulfonic acid (MES), histidine, ethanolamine, hydrazine, imidazole, pyridine and piperazine. Preferably, the amine group-containing buffering agent is selected from Tris and Bis-Tris.

[017] The buffer agent may be present in the composition at any concentration sufficient to maintain an ophthalmically-acceptable pH (e.g., 5.8 to 7.4). Typically, the buffer agent is present at a molar concentration of 0.001M-0.5M. Preferably, the buffer agent is present at a molar concentration of 0.005M-0.2M, more preferably 0.006M-0.15 M, most preferably 0.008-0.12M.

[018] The presence of the amine buffer increases the heat-stability of the muscarinic acetylcholine receptor antagonist so that decomposition of the muscarinic acetylcholine receptor antagonist during heat sterilization is reduced as compared to a buffered composition lacking an amine group-containing buffering agent, such as phosphate buffered saline. The heat-stability of the muscarinic acetylcholine receptor antagonist is further increased when it forms a complex with macrocyclic host molecule, for example a cyclodextrin or a cucurbituril. In addition, the amine buffer and the macrocyclic host molecule complex with the muscarinic acetylcholine receptor antagonist may increase the shelf life, as the buffering agent and macrocyclic host molecule prevents or reduces hydrolysis and / or degradation of the muscarinic acetylcholine receptor antagonist during storage. In some examples, the liquid composition remains stable when stored at room temperature (around 20°C) removing the need to refrigerate the liquid composition prior to or after first use. A liquid composition at ambient temperature is typically more comfortable to a patient when administered to the eye.

[019] As used herein, a “heat-stable” composition is one in which less than 10% (by weight) of the muscarinic acetylcholine receptor antagonist degrades during autoclave at 121 °C (at 1 atm) for 30 minutes at 17 psi (hereinafter, “autoclave”). Thus, a heat-stable liquid composition of the invention can be packaged in a container and autoclaved to provide a heat-sterilized container wherein the amount of muscarinic acetylcholine receptor antagonist in the liquid composition following autoclave is no less than 90% (by weight) of an initial amount of the muscarinic acetylcholine receptor antagonist present in the liquid composition prior to heat-sterilization. In some examples, the amount of muscarinic acetylcholine receptor antagonist in the liquid composition is no less than 95% of an initial amount of the muscarinic acetylcholine receptor antagonist present in the composition prior to heat-sterilization. In some examples, the liquid composition that has been packaged in a container and autoclaved comprises less than 10% hydrolysed degradation products of the muscarinic acetylcholine receptor antagonist, and preferably less than 5% hydrolysed degradation products of the muscarinic acetylcholine receptor antagonist.

[020] Preferably, the autoclaved composition comprises at least 50% (by weight) fewer muscarinic acetylcholine receptor antagonist degradation products than an autoclaved control composition. As used herein, a “control composition” is a composition comprising phosphate buffered saline (PBS) having the formulation provided in Table 1 below instead of the amine buffer and does not contain a macrocyclic host molecule but is otherwise identical. In other words, a control composition has the same pH and amounts of muscarinic acetylcholine receptor antagonist and excipients as the amine buffer and macrocyclic host molecule-containing composition to which it is being compared. Thus, for example, if a liquid composition comprising an amine buffer and 1.00 wt.% atropine is autoclaved and 0.95 wt.% atropine remains in the composition after autoclave, the autoclaved composition comprises 0.05 wt.% atropine degradation products. If a control composition (i.e., comprising 1.00 wt.% atropine in PBS buffer) contains 0.80 wt.% atropine after autoclave, the autoclaved composition comprising the amine buffer has 75% fewer degradation products than the control composition. The heat-stable composition may comprise at least 60%, 75% or 80% (by weight) fewer muscarinic acetylcholine receptor antagonist degradation products after autoclave than the control composition. The amount of degradation of a muscarinic acetylcholine receptor antagonist after autoclave can be determined by routine HPLC analysis, such as described in the examples below.

[021] While the compositions described herein are heat-stable, they may be sterilized using any known manner of sterilization, including by filtration methods typically used for ophthalmic formulations. In one example the composition is filtered through a 0.22 pm filter. The filter-sterilized, heat-stable composition may be stored and administered at room temperature, which can be more comfortable to the patient compared to the administration of ophthalmic formulations that require refrigeration.

[022] The muscarinic acetylcholine receptor antagonist may be heat sensitive. As used herein a “heat-sensitive muscarinic acetylcholine receptor antagonist” is a pharmaceutically active muscarinic acetylcholine receptor antagonist which is prone to degradation upon autoclave. In particular, less than 95% (by weight) of the initial amount of a heat-sensitive muscarinic acetylcholine receptor antagonist is present after autoclave when prepared as a 0.1% solution in PBS at pH 7.0. In some examples, less than 90% (by weight), or less than 80% (by weight), or less than 70% (by weight) of the initial amount of the heat-sensitive muscarinic acetylcholine receptor antagonist is present after autoclave.

[023] The muscarinic acetylcholine receptor antagonist comprises a hydrolysable bond. A hydrolysable bond is one which breaks through the process of hydrolysis. As used herein, a muscarinic acetylcholine receptor antagonist is considered to contain a hydrolysable bond if an aqueous solution of the muscarinic acetylcholine receptor antagonist contains at least 5% (by weight) breakdown products after autoclave. The hydrolysable bond may be an ester bond or an amide bond. In some examples, the hydrolysable bond is an ester bond or amide bond linked via one or more covalent bonds to an N-containing heterocyclic group. Examples of N-containing heterocyclic groups include substituted or unsubstituted aziridine, azirine, azetidine, azete, pyrrolidone, pyrrole, pyrimidine groups, imidazoline, pyrazolidine, imidazole, pyrazole, pyridine, piperidine, tetrazole, pentazole, diazine, diazinane, triazinane, triazine, tetrazine and tropane groups.

[024] The composition comprises a muscarinic acetylcholine receptor antagonist. Suitable muscarinic acetylcholine receptor antagonists include atropine, pirenzepine, scopolamine, trospium chloride, dicycloverine, oxybutynin, ipratropium bromide, and telenzepine. Preferably, the muscarinic acetylcholine receptor antagonist is atropine or pirenzepine or a derivative or analog thereof, and pharmaceutically acceptable salts thereof. As used herein “atropine” includes D-atropine, L-atropine (hyoscyamine) or a racemic mixture, or a pharmaceutically acceptable salt thereof. Suitable salts include hydrochloride, sulfate, acetate, phosphate, diphosphate, chloride, maleate, citrate, mesylate, nitrate, tartrate, maleate, bicarbonate fumarate, tosylate, succinate, stearate and gluconate. Preferably, the muscarinic acetylcholine receptor antagonist is atropine sulfate.

[025] In some examples, the composition comprises a muscarinic acetylcholine receptor antagonist of formula (I) or (II): Formula (I) Me Formula (II)

[026] in which A is O or NH;

[027] Ri is H, D (deuterium), hydroxyl, alkoxy, nitrile, a halogen atom, a Ci-Cio straight, branched or cycloalkyl group optionally substituted with one or more halogen atom; or a phenyl or benzyl group optionally substituted with one or more substituents selected from Ci-Cio straight, branched or cycloalkyl group, haloalkyl group, hydroxyl, alkoxy, nitrile, nitro, amino, amide, ester, sulfone, sulfoxide, sulfonamide, and halogen atoms; or a heterocyclic saturated, unsaturated or aromatic 5- or 6-member ring containing one or more heteroatoms selected from nitrogen, oxygen and sulfur and each being optionally substituted with one or more substituents selected from Ci- Cio straight, branched or cycloalkyl group, haloalky 1 group, hydroxyl, alkoxy, nitrile, nitro, amino, amide, ester, sulfone, sulfoxide, sulfonamide, and halogen atoms;

[028] X is -OH, O, or -ONO2;

[029] Me is CH3; and

[030] W is Nor CH.

[031] Preferably A is O.

[032] Preferably Ri is H, D or a halogen atom. Preferably the halogen atom is F.

[033] Preferably A is O and Ri is H, D or a halogen atom. Preferably the halogen atom is F.

[034] Preferably A is O and Ri is H.

[035] Preferably X is O.

[036] Preferably W is N.

[037] Preferably X is O and W is N.

[038] The composition may comprise 0.001 wt.% to 5.0 wt % muscarinic acetylcholine receptor antagonist. In some examples, the composition comprises 0.005 wt.% to 3.0 wt.%, or 0.01 wt.% to 2.0 wt.% muscarinic acetylcholine receptor antagonist.

[039] In the composition, the muscarinic acetylcholine receptor antagonist is present in a complex with a macrocyclic host molecule. A “macrocyclic host molecule” is a cyclic molecule with a cavity or pore-like structure which can bind or encapsulate some or all of a small guest molecule, such as the muscarinic acetylcholine receptor antagonist. The guest molecule is held in place through non-covalent bonds such as hydrophobic interactions, hydrogen bonding, van der Waals forces and / or electrostatic forces. Examples of suitable macrocyclic host molecules include cyclodextrins, cucurbiturils, crown ethers, cryptophanes, calixarenes, resorcinarenes and pyrogallolarenes.

[040] Preferred macrocyclic host molecules are cyclodextrins and cucurbiturils. Cyclodextrins are made up of glucose units held together by ether bonds. The number of glucose units determines the size of the cavity, a-cyclodextrin has 5 glucose units, P-cyclodextrin has 6 glucose units and y-cyclodextrin has 8 glucose units. The cyclodextrin can be chemically modified, for example, to improved solubility. Modified cyclodextrins include methyl-P-cyclodextrin, (2-hydroxyethyl)-P-cyclodextrin, 2- hydroxypropyl-P-cyclodextrin (HP-P-CD) and sulfobutylether-P-cyclodextrin (SBE-P-CD), carboxymethyl P cyclodextrin sodium salt, P cyclodextrin sulfate sodium salt and derivatives thereof. Preferred cyclodextrins are P-cyclodextrin and hydroxypropyl P-cyclodextrin.

[041] Cucurbiturils are made of glycoluril units linked by methylene bridges. Cucurbiturils are commonly written as cucurbit[n]uril, where n is the number of glycoluril units. Examples include cucurbit-6uril, cucurbit-7uril and cucurbit-8uril. A preferred cucurbituril is cucurbit-7uril.

[042] The muscarinic acetylcholine receptor antagonist and a macrocyclic host molecule are present in the complex in a 1:1 ratio. Therefore, the muscarinic acetylcholine receptor antagonist and the macrocyclic host molecule are present in the composition in at least equimolar amounts, i.e. at a 1:1 molar ratio. Preferably the muscarinic acetylcholine receptor antagonist and a macrocyclic host molecule are present in the formulation in the same amount between 50-5000 ppm.

[043] The composition may contain additional amounts of one or more uncomplexed macrocyclic host molecule so the total amount of macrocyclic host molecules present is greater than an equimolar amount to the muscarinic acetylcholine receptor antagonist. This means there are additional uncomplexed macrocyclic host molecules present, i.e., it is present in excess. The additional uncomplexed macrocyclic host molecule may be present at a concentration of 5-4000 ppm. Thus, the composition comprises 5-4000 ppm more macrocyclic host molecules than the amount of muscarinic acetylcholine receptor antagonist and macrocyclic host molecule used to form the complex. Preferably, the additional uncomplexed macrocyclic host molecule is present in an amount of 100-3000 ppm, more preferably 200-2500 ppm or 300-2250 ppm. Preferably, the additional uncomplexed macrocyclic host molecule is a cyclodextrin, and is present at a concentration of 1000 - 2000 ppm. Preferably, the additional uncomplexed macrocyclic host molecule is P-cyclodextrin, or hydroxypropyl P-Cyclodextrin. Preferably, the additional uncomplexed macrocyclic host molecule is a cucurbituril and is present at a concentration of 200 - 1000 ppm. Preferably, the additional uncomplexed macrocyclic host molecule is cucurbit-7uril.

[044] Preferably, the ratio of the total amount of macrocyclic host molecules present to the muscarinic acetylcholine receptor antagonist is in the range 1.25:1 to 4:1, more preferably 1.5:1 to 3:1.

[045] The composition may be prepared by mixing the muscarinic acetylcholine receptor antagonist and a macrocyclic host molecule together in an equimolar amount so that they form a complex. Additional macrocyclic host molecules may then be added so that they are present in a molar excess of the muscarinic acetylcholine receptor antagonist. The additional macrocyclic host molecule may be the same as or different to the macrocyclic host molecule used in the formation of the complex. Preferably, the additional uncomplexed macrocyclic host molecule is the same as the macrocyclic host molecule used in the formation of the complex. Preferably, the additional uncomplexed macrocyclic host molecule is a cyclodextrin or a cucurbituril. Preferably, the additional uncomplexed macrocyclic host molecule is P-cyclodextrin, hydroxypropyl P-cyclodextrin or cucurbit-7uril.

[046] The composition is ophthalmically-acceptable which refers to a composition which may be in direct contact with the ocular environment without causing any physiological or pharmacological harmful effects. Initial stinging or minor discomfort is common with topical ophthalmic formulations and is not considered harmful. The compositions disclosed herein have the advantage of reducing or eliminating the stinging or discomfort associated with previous ophthalmic formulations of muscarinic acetylcholine receptor antagonists.

[047] The use of heat sterilization, as opposed to filter sterilization removes the need for a preservative. Preferably, the composition does not contain a preservative, so the composition is preservative-free. In some examples, the composition comprises less than 0.001 wt% of a preservative. Examples of preservatives used in ophthalmic solutions include benzalkonium chloride, cetrimonium, sodium perborate, sodium chlorite, stabilized oxychloro complex, SofZia, polyquaternium-1, chlorobutanol, edetate disodium, polyhexamethylene biguanide, or combinations thereof.

[048] The presence of trace amounts of heavy metals in the buffer solution may accelerate the hydrolysis of muscarinic acetylcholine receptor antagonists which contain a hydrolysable bond. The composition may optionally further comprise a heavy metal chelator, such as the heavy metal chelator ethylenediaminetetraacetic acid (EDTA). The amount of the chelator, e.g., EDTA present in the composition may be from about 10 ppm to 5000 ppm, preferably from about 100 ppm to about 1000 ppm, and more preferably from about 100 ppm to about 500 ppm. In one example the composition comprises 150 ppm EDTA.

[049] Where the composition comprises a molar excess of the macrocyclic host molecule, it preferably also includes EDTA. In this preferred embodiment, the molar ratio of total macrocyclic host molecule: muscarinic acetylcholine receptor antagonist present is >1:1, and the composition further comprises EDTA. Preferably, the ratio of the total amount of macrocyclic host molecules present to the muscarinic acetylcholine receptor antagonist is in the range 1.25: to 4:1, more preferably 1.5:1 to 3:1, and further comprises EDTA. The composition preferably comprises from about 10 ppm to about 5000 ppm EDTA, preferably about 100 ppm to about 1000 ppm EDTA, and more preferably from about 100 ppm to about 500 ppm EDTA. Preferably, the additional uncomplexed macrocyclic host molecule may be present at a concentration of 250-3000 ppm, more preferably 300-2500 ppm or 750-2250 ppm. Even more preferably, the additional uncomplexed macrocyclic host molecule is present at a concentration of 1000 - 2000 ppm. In one preferred composition, the additional uncomplexed macrocyclic host molecule is present at an amount of 50-4000 ppm and the EDTA is present in an amount of 10 ppm to 5000 ppm. Most preferably, the composition comprises 300-2250 ppm additional uncomplexed macrocyclic host molecule and 100 ppm to 500 ppm EDTA, most particularly 150 ppm EDTA. Preferably, the additional uncomplexed macrocyclic host molecule is a cyclodextrin or a cucurbituril. Preferably, the additional uncomplexed macrocyclic host molecule is 0-cyclodextrin, hydroxypropyl 0-cyclodextrin or cucurbit-7uril.

[050] The composition may optionally further comprise an agent capable of increasing intermolecular hydrogen bonding. These agents may help to further stabilize the muscarinic acetylcholine receptor antagonist after autoclave resulting in fewer muscarinic acetylcholine receptor antagonist degradation products. Preferably, the presence of the agent capable of increasing intermolecular hydrogen bonding results in at least 5% (by weight) fewer muscarinic acetylcholine receptor antagonist degradation products than an otherwise identical control composition without the agent capable of increasing intermolecular hydrogen bonding following autoclave. Examples of agents capable of increasing intermolecular hydrogen bonding include trehalose, ethylene glycol, and mannitol. Preferably, the agent capable of increasing intermolecular hydrogen bonding is trehalose.

[051] The agent capable of increasing intermolecular hydrogen bonding may be present in the composition in an amount of about 0.1 wt % to about 10.0 wt % (based on the total weight of the composition). Preferably, the composition comprises from about 0.5 wt % to about 5.0 wt % agent capable of increasing intermolecular hydrogen bonding, more preferably from about 1.0 wt % to about 4.0 wt %, even more preferably from about 2.0 wt % to about 3.0 wt %. In some examples, the composition may comprise 2.0 wt. % trehalose.

[052] Intermolecular hydrogen bonding can also be increased by formulating the composition with deuterated water (D2O) instead of regular water. Thus, in some examples, the heat stable ophthalmically-acceptable aqueous liquid composition comprises an amine buffer and a muscarinic acetylcholine receptor antagonist which comprises a hydrolysable bond, wherein the aqueous component of the composition consists of or comprises deuterated water.

[053] The liquid composition has an ophthalmically-acceptable osmolarity. An ophthalmically-acceptable osmolarity is in the range of about 200 mOsm / kg to about 500 mOsm / kg, and typically from about 270 mOsm / kg up to about 330 mOsm / kg.

[054] The liquid composition optionally comprises a tonicity agent in an amount to maintain an ophthalmically-acceptable osmolarity, such as about 0.1 wt.% to 5.0 wt.%, or about 0.5 wt.% to about 3.0 wt.%. Suitable tonicity agents include 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.

[055] The liquid composition optionally comprises an excipient that extends the period of time that a dose of the composition remains in contact with the cornea. For example, the excipient may be a viscosity-enhancing agent and / or a mucoadhesive agent. Suitable viscosity-enhancing agents include cellulose-based polymers (e.g., hydroxypropyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose, and hydroxyethyl cellulose), polyoxyethylene-polyoxypropylene triblock copolymers (e.g., Poloxamer 407), dextran-based polymers, polyvinyl alcohol, dextrin, polyvinylpyrrolidone, polyalkylene glycols, chitosan, collagen, gelatin, gellan gum, xanthan gum, guar gum, tamarind seed polysaccharide, hyaluronic acid, or combinations thereof.

[056] The liquid composition may optionally contain additional ingredients such as a comfort agent, a hydrophilic polymer, a wetting agent, a surfactant or other additive.

[057] The invention also provides a container containing a heat-stable ophthalmically-acceptable liquid composition as described herein. In some examples the container is heat-sterilized. In other examples the container is not heat-sterilized, and the ophthalmically-acceptable liquid composition is filter-sterilized. The container may be an eye drop dispenser such as an eye drop dispensing bottle or vial. The dispenser may be single use or multiple use. Some eye drop dispensers have a nozzle connected to a receptacle containing the liquid composition. The receptacle may have deformable walls so that the user can squeeze the receptacle to force the liquid composition into the nozzle. The nozzle allows a drop of the composition to be administered in the lower conjunctival sac and thus to the surface of the eye. Alternatively, a multidose dispenser may comprise a bottle containing the liquid composition of the invention, a tip or dropper, and a closure cap. The bottle may be rigid, for example, made from glass or a rigid plastic. The bottle may have a removable pipette with a deformable bulb to control movement of the liquid within the dropper.

[058] The receptacle can typically hold a volume of 0.01 ml to 20 ml. Single use dispensers typically contain sufficient fluid to provide the dosage required by a patient at one time. This may be a single drop for a single eye, or multiple drops (e.g., 2 or 3) for one or both eyes. Single use dispensers typically hold a volume of 0.01 ml to 0.1 ml. Multiple use dispensers contain multiple doses for administration a number of times, and typically hold a volume of 1.0 ml to 20 ml. Preferably, the multiple use dispenser is compatible for use with devices designed to aid the instilling of eye drops.

[059] The container may be a contact lens package. The container may comprise a base member comprising a bowl that defines a blister cavity dimensioned to accommodate a contact lens and the liquid composition and a flange projecting away from the blister cavity. The base member can be formed from any suitable material, such as glass or a thermoplastic material. A liquid-tight seal can be provided by a cover attached to the flange region, such as a removable foil that is glued or heat sealed to the flange region. Such contact lens containers (i.e., packages), which are commonly referred to as “blister packs”, are well-known in the art (see e.g., U.S. Pat. No. 7,426,993). Generally, the container is configured to receive a single contact lens and an amount of liquid composition sufficient to completely cover the contact lens, typically about 0.5-1.5 ml.

[060] The container can also be a “foil to foil package” which comprises two foil members bonded together, which may either be flat, or one or both foil members formed into an appropriate geometric shape to accommodate contact the lens and liquid composition. In another example, the container is in the form of a plastic base member comprising a plurality of threads and the cover comprises a plastic cap member comprising a compatible set of threads for engagement with the threads of the base member thereby providing a resealable cover. It will be appreciated that other types of containers can be used.

[061] The sealed contact lens package may be sterilized by sterilizing amounts of radiation, including heat or steam, such as by autoclaving. Preferably, the packaged contact lens is sterilized by autoclaving. Autoclaving contact lens packages generally involves subjecting the sealed contact lens package to temperatures of at least 121°C for at least 30 minutes at 17 psi (117 kPa).

[062] The liquid composition of the invention can be used to treat an ophthalmic disorder. Preferably, the ophthalmic disorder or condition is pre-myopia, myopia, or progression of myopia. The invention also provides an ophthalmically-acceptable liquid composition for use in medicine. Specifically, the ophthalmically-acceptable liquid composition can be used in the treatment of an eye disorder, such as myopia.

[063] The ophthalmically-acceptable liquid composition is formulated for administration onto the surface of the eye. It is not formulated for administration via injection.

[064] The following Examples illustrate certain aspects and advantages of the present invention, which should be understood not to be limited thereby.

[065] Example 1: Synthesis of macrocyclic host molecule complexes with atropine

[066] Complex of atropine and B-cyclodextrin (B-CD): 10.0 g B-CD was added to 600 mL DI water in a 1 L reactor. 2.05 g atropine was dissolved in 50 mL ethanol, then added into the reactor. The mixture was stirred at room temperature for 24 hr. The precipitates formed were separated by filtration, rinsed with ethanol and vacuum dried to produce 8.58 g white solid.

[067] Complex of atropine and (2-hvdroxypropyl) B-cyclodextrin (HPCD)

[068] 15.41 g HPCD was added with 3.18 g atropine and 80 mL ethanol into a 250 mL reactor. The mixture was stirred at room temperature for 24 hr. The clear solution was evaporated to removed the alcohol. The residue was rinsed with acetone and vacuum dried to get 17.54 g white solid.

[069] Complex of atropine and cucurbit7uril (CB7): 4.0 g CB7 was added to 800 mL DI water in a 1 L reactor. 2.05 g atropine was dissolved in 40 mL water, and the pH adjusted with IN HC1 to pH 2.5, then added into the reactor. The mixture was kept at pH 4.0 and stirred at room temperature for 24 hr. The solution was neutralized with IN NaOH to pH 7 and freeze dried with lyophilizer. The collected precipitate was washed with 150 mL acetone and dried to get 4.39 g white solid.

[070] The amount of atropine in the obtained complex was quantified as described HPLC method in Table 2.

[071] Example 2. Effect of encapsulation with macrocyclic host molecule on autoclavestability of atropine

[072] A study was conducted to evaluate the stability of atropine in the amine containing BisTris buffer after autoclave at pH 6~7. The buffer formulation is provided below in Table 1.

[073] TABLE 1: Buffer solutions Phosphate Buffered Saline (PBS) Ingredient Composition wt% Sodium Chloride 0.830% Sodium Phosphate Monobasic 0.071% Sodium Phosphate Dibasic 0.599% Water 98.500% Bis-Tris Solution Ingredient Composition wt% Sodium Chloride 0.894% Bis (2-hydroxy ethyl) amino-tris(hydroxymethyl)methane 0.050% Water 99.056%

[074] Sample solutions preparation: Atropine and atropine / B-CD complex were dissolved in the buffer to obtain 0.1% w / w solutions. Atropine / CB7 complex was dissolved in the buffer solution to obtain a 0.07% w / w solution. The pH of each solution was adjusted to the values shown in Table 3 using HC1 or NH40H.

[075] 4 mL of each solution was added to 6 mL glass vials and sealed with a cap. The vials were autoclaved at 121° C for 30 minutes at 17 psi (117 kPa).

[076] Reverse phase HPLC was used to detect the presence of racemic atropine in the samples pre-autoclave and post-autoclave samples using the conditions shown in Table 2.

[077] TABLE 2: HPLC conditions Parameter Reverse phase HPLC Column Atlantis T3 5 pm 4.6x250 mm Part#l 86003748 with Guard column Temperature 30°C Mobile phase Trifluoroacetic acid in water (0.05% v / v)-acetonitrile (80-20) Elution rate 1 mL / min Detector UV at 256 nm

[078] The percentage of atropine degradation from the autoclaved samples compared to the pre autoclaved samples is shown in Table 3.

[079] TABLE 3: Stability of atropine and its CD or CB7 complex after autoclave Test Compound Bis-Tris buffer pH % Atropine decomposition post-AC Atropine 6.27 2.4% Atropine / B-CD complex 6.23 1.5% Atropine / CB7 complex 6.20 0

[080] Example 3: Shelf-Life Stabilities of Atropine Complexes With B-CD, B-HPCD Are Evaluated with EDTA And Excess CD Or B-HPCD as Additives

[081] Eight test solutions as listed in Table 4 were prepared by dissolving the components in BisTris pH ~3.0. the pH was then adjusted to -pH 6.0 separately with HC1 or NaOH solution. A control solution was prepared by dissolving 1% (racemic)atropine w / w in 0. IN HC1, before diluting to 0.01% concentration with Bis-Tris buffer. The pH was adjusted to pH 6.0-6.5 separately with HC1 or NaOH solution.

[082] TABLE 4: Test solutions Solution ID Atropine / B-CD complex (%w / w) Atropine / B-HPCD complex (%w / w) EDTA (%w / w) B-CD (%w / w) B-HPCD (%w / w) 1 0.1 0 0 0 0 2 0.1 0 0.015 0 0 3 0.1 0 0 0.1 0 4 0.1 0 0.015 0.1 0 5 0 0.1 0 0 0 6 0 0.1 0.015 0 0 7 0 0.1 0 0 0.2 8 0 0.1 0.015 0 0.2

[083] 4 mL of each solution was added to 6 mL glass vials and sealed with a cap. Four sets of vials were prepared for each solution - Pre autoclave, Time zero, 6 months equivalent and 1-year equivalent samples. The time zero, 6 month and 1-year sample vials were autoclaved at 121° C for 30 minutes at 17 psi (117 kPa). The pre-autoclave and time zero samples were tested to measure the amount of atropine degradation. Pre-autoclave and time zero samples were also stored in freezer for later comparison to 35°C samples. The 6 month and 1-year samples were kept in 35°C oven protected from light. After 90 days, the 6-month samples were retrieved and placed in the freezer. After 180 days the 1 -year samples were retrieved and placed in the freezer. All the frozen samples were then tested to determine atropine decomposition rates.

[084] Reverse phase HPLC was used to detect the presence of atropine using the conditions shown in Table 2 above. The results are summarized in Table 5.

[085] TABLE 5: Decomposition rate of atropine complexed with B-CD or B-HPCD after autoclave under storage at 3 5 °C Solution ID Shelf-life time Tested pH % Degradation of atropine 1 Post-autoclave (TO) 6.23 1.5% RT Eq 1-year 6.25 14.9% 2 Post-autoclave (TO) 6.06 0.0% RT Eq 1-year 6.15 18.5% 3 Post-autoclave (TO) 6.15 0% RT Eq 1-year 6.39 11.6% 4 Post-autoclave (TO) 6.02 1.8% RT Eq 0.5-year 6.07 1.8% RT Eq 1-year 5.93 7.5% 5 Post-autoclave (TO) 6.10 4.1% RT Eq 1-year 6.15 17.7% 6 Post-autoclave (TO) 5.93 1.8% RT Eq 1-year 6.25 17.1% 7 Post-autoclave (TO) 6.38 0% RT Eq 1-year 6.51 17.4% 8 Post-autoclave (TO) 5.88 1.9% RT Eq 0.5-year 6.05 5.1% RT Eq 1-year 6.02 9.9%

[086] The test results for the control atropine solution after autoclave under storage at 35°C are summarized in Table 6.

[087] TABLE 6: Decomposition rate of atropine after autoclave under storage at 35°C Shelf-life time Target pH Test pH % Degradation of atropine Post-autoclave (TO) 6.0 6.27 2.4% RTEq 1-year 6.13 14.5% RT Eq 2-year 6.10 31.2% Post-autoclave (TO) 6.5 6.69 3.7% RTEq 1-year 6.54 26.5% RT Eq 2-year 6.49 60.0%

[088] Comparing the above data it is observed that the combination of EDTA and excess CD or HPCD has a synergistic effect on stabilization of the atropine complex with CD or HPCD, with an atropine decomposition rate of-10% RT equivalent 1-year. Meanwhile, atropine was decomposed -14.5% at pH 6.13 and -26.5% at pH 6.54 without additives under similar test condition.

[089] Example 4: Evaluation of the shelf-life of Atropine / CB7 complex with EDTA, CD and excess CB7 as additives in Bis-Tris at pH 6-6.5

[090] Solutions preparation. The test solutions listed in Table 7 were prepared by dissolving the components in Bis-Tris pH -3.0. the pH was then adjusted to pH 6.0-6.5 separately with HC1 or NaOH solution. A control solution was prepared by dissolving 1% atropine w / w in 0. IN HC1, before diluting to 0.01% concentration with Bis-Tris buffer. The pH was adjusted to pH 6.0-6.5 separately with HC1 or NaOH solution.

[091] TABLE 7: Test solutions. Solution ID Atropine / CB7 (%w / w) B-CD (%w / w) CB7 (%w / w) EDTA (%w / w) PH 9 0.07 0 0 0 6.0 10 0.07 0 0 0.015 6.0 11 0.07 0.1 0 0 6.0 12 0.07 0.1 0 0.015 6.0 13 0.07 0 0.03 0 6.0 14 0.07 0 0.03 0.015 6.0 15 0.07 0 0 0 6.5 16 0.07 0 0 0.015 6.5 17 0.07 0.1 0 0 6.5 18 0.07 0.1 0 0.015 6.5 19 0.07 0 0.03 0 6.5 20 0.07 0 0.03 0.015 6.5

[092] 3 mL of each solution was added to 6 mL glass vials and sealed with a cap. Five sets of vials were prepared for each solution: Pre-autoclave, Time zero, 6 months equivalent, 1-year equivalent and 2-year equivalent samples. The time zero, 6 month, 1-year and 2-year sample vials were autoclaved at 121° C for 30 minutes at 17 psi (117 kPa). The pre-autoclave and time zero samples were tested to measure the amount of atropine degradation. Pre-autoclave and time zero samples were also stored in a freezer for later comparison to 35°C samples. The 6 month, 1-year and 2-year samples were kept in a 35°C oven protected from light. After 90 days, the 6-month samples were retrieved and placed in the freezer. After 180 days the 1-year samples were retrieved and placed in the freezer. After 360 days the 2-year samples were retrieved and placed in the freezer. All the frozen samples were then tested to determine atropine decomposition rates. Reverse phase HPLC was used to detect the presence of atropine using the conditions shown in Table 2 above. The test results for atropine / CB7 complex at 35°C are summarized in Table 8.

[093] TABLE 8. Decomposition rate of atropine / CB7 complex after autoclave in Bis-Tris / pH ~6 and storage at 35°C Solution ID Shelf-life time Test pH % Degradation of atropine 9 Post-autoclave (TO) 6.20 0% RT Eq 1-year 6.13 1.7% RT Eq 2-year 6.25 17.0% 10 Post-autoclave (TO) 6.02 1.0% RT Eq 1-year 6.22 7.5% RT Eq 2-year 6.20 16.0% 11 Post-autoclave (TO) 6.26 0% RT Eq 1-year 6.17 3.2% RT Eq 2-year 6.27 15.8% 12 Post-autoclave (TO) 6.23 2.7% RT Eq 1-year 6.15 6.5% RT Eq 2-year 6.18 14.3% 13 Post-autoclave (TO) 5.89 6.3% RT Eq 1-year 5.92 4.6% RT Eq 2-year 6.00 6.6% 14 Post-autoclave (TO) 5.81 3.5% RT Eq 1-year 5.81 0.3% RT Eq 2-year 5.91 5.0% Solution ID Shelf-life time Test pH % Degradation of atropine 15 Post-autoclave (TO) 6.43 0% RT Eq 1-year 6.34 4.5% RT Eq 2-year 6.28 19.6% 16 Post-autoclave (TO) 6.33 0% RT Eq 1-year 6.33 8.6% RT Eq 2-year 6.42 26.3% 17 Post-autoclave (TO) 6.37 0% RT Eq 1-year 6.25 8.1% RT Eq 2-year 6.35 18.0% 18 Post-autoclave (TO) 6.37 0% RT Eq 1-year 6.39 9.9% RT Eq 2-year 6.46 23.8% 19 Post-autoclave (TO) 6.40 6.1% RT Eq 1-year 6.11 5.8% RT Eq 2-year 6.17 9.2% 20 Post-autoclave (TO) 6.00 3.8% RT Eq 1-year 6.23 3.2% RT Eq 2-year 6.18 8.7%

[094] By comparison of the above data with the atropine control data in Table 6, it is observed that encapsulation of atropine with CB7 has a stabilizing effect. The decomposition rates at pH 6~6.5 attain lower than 10% degradation after RT equivalent 2-year while uncomplexed atropine decomposed 14.5% at pH ~6 after RT equivalent 1-year, and 31.2% after RT equivalent 2-year.

[095] The disclosure herein refers to certain illustrated examples, it is to be understood that these examples are presented by way of example and not by way of limitation. The intent of the foregoing detailed description, although discussing exemplary examples, is to be construed to cover all modifications, alternatives, and equivalents of the examples as may fall within the spirit and scope of the invention as defined by the additional disclosure.

[096] Applicants specifically incorporate the entire contents of all cited references in this disclosure. Further, when an amount, concentration, or other value or parameter is given as either a range, preferred range, or a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. Where a range of numerical values is recited herein, unless otherwise stated, the range is intended to include the endpoints thereof, and all integers and fractions within the range. It is not intended that the scope of the invention be limited to the specific values recited when defining a range.

[097] The present invention can include any combination of the various features or embodiments described above and / or in the claims below as set forth in sentences and / or paragraphs. Any combination of disclosed features herein is considered part of the present invention and no limitation is intended with respect to combinable features.

[098] Other embodiments of the present invention will be apparent to those skilled in the art from consideration of the present specification and practice of the present invention disclosed herein. It is intended that the present specification and examples be considered as exemplary only with a true scope and spirit of the invention being indicated by the following claims and equivalents thereof.

Claims

1. An ophthalmically-acceptable aqueous liquid composition comprising an amine buffer and a muscarinic acetylcholine receptor antagonist which comprises a hydrolysable bond complexed with a macrocyclic host molecule.

2. The ophthalmically-acceptable liquid composition of claim 1, having a pH in the range of 5.0 to 7.5.

3. The ophthalmically-acceptable aqueous liquid composition of claim 1 or claim 2, having a pH of 5.8 to 7.4.

4. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the hydrolysable bond is an ester bond or an amide bond linked to a nitrogen-containing heterocyclic group.

5. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the muscarinic acetylcholine receptor antagonist is atropine or pirenzepine.

6. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the macrocyclic host molecule is a cyclodextrin or cucurbituril.

7. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the macrocyclic host molecule is P-cyclodextrin or hydroxypropyl P-cyclodextrin.

8. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the macrocyclic host molecule is cucurbit[7]uril.

9. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the composition is preservative-free.

10. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the composition further comprises EDTA and additional uncomplexed macrocyclic host molecule.

11. The ophthalmically-acceptable aqueous liquid composition of claim 10, wherein the additional uncomplexed macrocyclic host molecule is the same macrocyclic host molecule complexed with the muscarinic acetylcholine receptor antagonist.

12. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the amine buffer is selected from Tris(hydroxymethyl)aminomethane (Tris), N-(2-Acetamido)iminodiacetic acid (ADA), bis(2-Hydroxyethyl)iminotris(hydroxymethyl)methane (Bis-Tris), 2-morpholinoethanesulfonic acid (MES), histidine, ethanolamine, hydrazine, imidazole, pyridine and piperazine.

13. The ophthalmically-acceptable aqueous liquid composition of any preceding claim, wherein the amine buffer is selected from Tris and Bis-Tris.

14. A heat-sterilized container containing the ophthalmically-acceptable aqueous liquid composition of any one of claims 1 to 13.

15. The heat-sterilized container of claim 14, wherein the liquid composition comprises less than 10% hydrolysed degradation products of the muscarinic acetylcholine receptor antagonist.

16. The heat-sterilized container of claim 14 or claim 15, wherein the container is a multi-use or single-use eye drop dispensing bottle or vial.

17. The heat-sterilized container of any one of claims 14 to claim 16, wherein the container further contains a contact lens.

18. The heat-sterilized container of any one of claims 14 to 17, wherein the amount of muscarinic acetylcholine receptor antagonist in the composition is no less than 90% of an initialamount of the muscarinic acetylcholine receptor antagonist present in the composition prior to heat-sterilization.

19. The heat-sterilized container of any one of claims 14 to 18, wherein the composition comprises at least 90% (by weight) fewer hydrolysed degradation products of the muscarinic acetylcholine receptor antagonist than a control composition comprising PBS instead of the amine buffer.

20. The heat-sterilized container of any one of claims 14 to 19, wherein the heat-sterilized container is sterilized by autoclaving.

21. A method of manufacturing the heat-sterilized container of any one of claims 14 to 20 comprising:(a) placing an ophthalmically-acceptable solution in a container,(b) sealing the container, and(c) heat sterilizing the sealed container, wherein the solution is a liquid ophthalmically acceptable composition as defined in any one of claims 1 to 13.

22. A container containing the ophthalmically-acceptable aqueous liquid composition of any one of claims 1 to 13, wherein the liquid composition has been sterilized by filtration.

23. The container of claim 22, wherein the container is a multi-use or single-use eye drop dispensing bottle or vial.T +44(0)30 0300 2000A