Pharmaceutical compositions for intraocular administration

Abstract

Provided is a pharmaceutical composition that can be easily administered in the ear and has the function of retaining a medication and sustainedly releasing the same in the ear. This pharmaceutical composition for otic administration comprises one or more kinds of medications and a water-dispersible microcellulose composition, in particular, crystalline cellulose carmellose sodium.

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition that can be easily administered into the ear and has the function of retaining and sustained-release of a drug in the ear. [Background technology]

[0002] There are still few drug treatments for ear diseases, and surgery is often relied upon. Even if an effective drug is discovered, oral or intravenous administration can result in reduced efficacy or side effects. Therefore, ear drops are primarily used as pharmaceutical agents or pharmaceutical compositions for local administration of drugs to or near the affected ear site, and the development of technologies for drug retention and sustained release is anticipated.

[0003] Patent Document 1 (WO2011 / 049958) reports a sustained-release pharmaceutical formulation that utilizes a thermosensitive polymer. When a solution of a thermosensitive polymer, polyoxyethylene-polyoxypropylene copolymer, dissolved at about 5% to about 20% by weight is administered into the ear via a syringe needle, it gels due to body temperature (Non-Patent Document 1: James M. Chamberlain et al., ANNALS OF EMERGENCY MEDICINE, 1995, 25(1), pp. 15-20, reports that the temperature inside the ear canal is about 37°C to about 38°C). Therefore, it is reported that the formulation can be applied to the round window membrane.

[0004] Patent Document 2 (WO2014 / 186075) reports that biodegradable chitosan-based hydrogels can deliver drugs and be injected into the ear canal. The hydrogel formulations can solidify within minutes after mixing with a catalyst for chemical crosslinking, can be easily administered using a dual-bore syringe, or can be mixed with a crosslinker before delivery and injected using a single-bore syringe.

[0005] Although several pharmaceutical compositions capable of retaining and sustaining the drug in the ear have been reported, there is room for further study to provide a pharmaceutical composition that can be easily administered in the ear and has the function of retaining and sustaining the drug in the ear, due to the need for strict temperature control, limited administration time, or special administration devices. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Publication No. 2011 / 049958 [Patent Document 2] International Publication No. 2014 / 186075 [Non-patent literature]

[0007] [Non-Patent Document 1] James M Chamberlain et al.,ANNALS OF EMERGENCY MEDICINE,1995,25(1),p.15-20 Summary of the Invention [Problem to be solved by the invention]

[0008] An object of the present invention is to provide a pharmaceutical composition that can be easily administered intraearly and has the function of retaining and sustained-release of a drug in the ear. [Means for solving the problem]

[0009] The present inventors have conducted extensive research into pharmaceutical compositions that can be easily administered intraearly and that have the function of retaining and sustaining the drug in the ear. As a result, they have discovered that a pharmaceutical composition containing a water-dispersible fine cellulose composition has a viscosity that is not affected by temperature under temperature conditions ranging from room temperature to the temperature inside the ear canal, does not require a special administration device, has a low maximum administration pressure when injected, retains the drug in the ear, and has sustained drug release properties, and have thus completed the present invention.

[0010] That is, the present invention is [1] A pharmaceutical composition for intraaural administration containing one or more drugs and a water-dispersible fine cellulose composition; [2] The pharmaceutical composition according to [1], wherein the water-dispersible fine cellulose composition contains crystalline cellulose and a water-soluble polymer. [3] The pharmaceutical composition according to [2], wherein the water-soluble polymer is carmellose sodium. [4] The pharmaceutical composition according to [1], wherein the water-dispersible fine cellulose composition is crystalline cellulose-carmellose sodium. [5] Any of the pharmaceutical compositions according to [1] to [4], further comprising a solvent. [6] The pharmaceutical composition according to [5], wherein the solvent is water. [7] The pharmaceutical composition according to [5] or [6], wherein the total weight ratio of the water-dispersible fine cellulose composition to the pharmaceutical composition is 1.7% (w / w) to 9.1% (w / w). [8] The pharmaceutical composition according to [5] or [6], wherein the total weight ratio of the water-dispersible fine cellulose composition to the pharmaceutical composition is 2.9% (w / w) to 9.1% (w / w). [9] Any of the pharmaceutical compositions [1] to [8], which sustainably releases a drug when an insert with a permeable membrane is fitted into a plate well containing 2 mL of phosphate buffered saline at 37°C ± 2°C so as to separate the phosphate buffered saline into an upper layer and a lower layer, 0.5 mL of any of the pharmaceutical compositions [1] to [8] is poured onto the insert, and the plate well is shaken at a speed that causes the water surface of the phosphate buffered saline to sway, thereby dissolving the drug.

[10] Any of the pharmaceutical compositions according to [1] to [9], further containing a solubilizing agent.

[11] The pharmaceutical composition according to any one of [1] to

[10] , wherein the drug is one or more selected from the group consisting of low molecular weight compounds, nucleic acids, and proteins.

[12] The pharmaceutical composition according to any one of [1] to

[11] , wherein the drug is a drug that provides the biological activity of heparin-binding epidermal growth factor.

[13] The pharmaceutical composition according to

[12] , wherein the drug providing the biological activity of heparin-binding epidermal growth factor comprises a protein consisting of the amino acid sequence shown in SEQ ID NO: 1.

[14] A method for retaining a pharmaceutical composition for intraaural administration containing one or more drugs in the ear using a water-dispersible fine cellulose composition.

[15] The method according to

[14] , wherein the water-dispersible fine cellulose composition is crystalline cellulose-carmellose sodium. Regarding. [Effects of the Invention]

[0011] According to the present invention, a pharmaceutical composition can be provided which contains one or more drugs and a water-dispersible fine cellulose composition, particularly crystalline cellulose-carmellose sodium, and which can be easily administered intraearly and has the function of retaining and sustained-release of the drug in the ear. Furthermore, the present invention provides a method for retaining a pharmaceutical composition for intraaural administration containing one or more drugs in the ear by using a water-dispersible fine cellulose composition, particularly crystalline cellulose-carmellose sodium. [Brief explanation of the drawings]

[0012] [Figure 1] 1 is a graph showing the results of a flowability test of a microcrystalline cellulose·carmellose sodium (MCC·CMCNa) base liquid and a Pluronic® F-127 (Plu.) base liquid conducted in Test Example 2, in which the distance traveled down the bottom of a stainless steel square tray tilted at 30 degrees was measured every 10 seconds for 60 seconds. [Figure 2] 1 is a graph showing the results of static viscosity and dynamic viscosity measurements of an MCC·CMCNa base solution carried out in Test Example 4-2 to evaluate the effect of base concentration. [Figure 3] 1 is a graph showing the results of static viscosity and dynamic viscosity measurements of an MCC·CMCNa base solution carried out in Test Example 5-1 to evaluate the effect of DMSO on the viscosity of the base solution. [Figure 4]1 is a graph showing the results of a complex viscosity measurement test of a Plus base solution conducted in Test Example 5-2 to evaluate the effect of the solubilizing agent dimethyl sulfoxide (DMSO) on the viscosity of the base solution. DETAILED DESCRIPTION OF THE INVENTION

[0013] The present invention relates to a pharmaceutical composition for intraaural administration, which contains one or more drugs and a water-dispersible fine cellulose composition, particularly microcrystalline cellulose-carmellose sodium.

[0014] The ear is divided into the outer ear, middle ear, and inner ear. The outer ear consists of the pinna, ear canal, and outer membrane of the eardrum. The middle ear consists of the inner membrane of the eardrum, ossicles (malleus, incus, stapes), tympanic cavity, ossicular muscles, Eustachian tube, mastoid sinus, and mastoid acinus. The inner ear consists of the oval (vestibular) window, vestibule, semicircular canals, utricle, saccule, round (cochlear) window, and cochlea.

[0015] As used herein, "aural administration" typically refers to, but is not limited to, administering a pharmaceutical composition into the external auditory canal, near the outer membrane of the tympanic membrane, on the tympanic membrane, near the inner membrane of the tympanic membrane, inside the tympanic cavity, on the round window membrane, or near the round window. Preferably, the pharmaceutical composition is administered near the inner membrane of the tympanic membrane, inside the tympanic cavity, on the round window membrane, or near the round window. Furthermore, a "pharmaceutical composition for intraaural administration" typically refers to, but is not limited to, a pharmaceutical composition for administration into the external auditory canal, near the outer membrane of the tympanic membrane, on the tympanic membrane, near the inner membrane of the tympanic membrane, inside the tympanic cavity, on the round window membrane, or near the round window. Preferably, the pharmaceutical composition is for administration near the inner membrane of the tympanic membrane, inside the tympanic cavity, on the round window membrane, or near the round window.

[0016] Typical forms of the pharmaceutical composition of the present invention include, but are not limited to, solid, powder, liquid, paste, etc. Liquid is preferred. In this specification, sol, gel, suspension, dispersion, and lotion can be included in the liquid, with dispersion being the preferred liquid. In this specification, sol means a fluid state in which colloidal particles are dispersed in a liquid, and gel means a state in which the viscosity of the sol has increased and the fluidity has been lost. When the pharmaceutical composition is in a liquid form and contains a drug, it may be called a "liquid."

[0017] In the present invention, "water-dispersible fine cellulose composition" refers to a composition containing crystalline cellulose that is easily dispersed in water. The water-dispersible fine cellulose composition used in the present invention typically includes, but is not limited to, a composition consisting of a mixture in which a water-soluble polymer is attached to or coated on crystalline cellulose. Preferably, the composition is a mixture in which a water-soluble polymer is coated on crystalline cellulose. Examples of methods for attaching or coating a water-soluble polymer to crystalline cellulose include, for example, mixing crystalline cellulose and a water-soluble polymer, adding a solvent such as purified water to form a suspension or paste, spreading the suspension or paste thinly, drying by heat treatment, etc., and then pulverizing it to form a powder; or a method in which a solution in which a water-soluble polymer is dissolved is sprayed onto crystalline cellulose fluidized with a heated air current, drying, and then sizing the cellulose.

[0018] In the present invention, the term "water-soluble polymer" refers to a polymer that has many hydrophilic polar groups along the polymer backbone and therefore dissolves in water as a macromolecule. The water-soluble polymer used in the present invention is not particularly limited as long as it is pharmaceutically acceptable and enhances the dispersibility of crystalline cellulose in water. Typical examples of water-soluble polymers include natural plant polymers such as guar gum, carrageenan, karaya gum, locust bean gum, gellan gum, glucomannan, sodium alginate, and corn starch; natural microbial polymers such as xanthan gum; natural animal polymers such as sodium chondroitin sulfate and sodium hyaluronate; semi-synthetic polymers such as carmellose sodium, dextrin, methylcellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, and cationized guar gum; and synthetic polymers such as carboxyvinyl polymers, polyacrylic acid, polyvinylpyrrolidone, and polyvinyl alcohol. Preferred examples include carmellose sodium, karaya gum, dextrin, and xanthan gum, and more preferred examples are carmellose sodium.

[0019] Carmellose sodium, sometimes called carboxymethylcellulose sodium or CMC sodium (hereinafter sometimes referred to as CMCNa), is a water-soluble cellulose polymer. Carmellose sodium can be distinguished by its viscosity (25°C, 60 rpm) and degree of etherification (degree of substitution) when made into a 1% aqueous solution. The viscosity is typically 20 to 500 Pa·s, preferably 50 to 300 Pa·s, and the degree of substitution is typically 0.5 to 2.0, preferably 0.55 to 1.1. Preferred commercial products of carmellose sodium include, but are not limited to, CMC Daicel 1120, 1130, 1140, 1150, and 1160 (manufactured by Daicel) and Sunrose® F-10MC and F-30MC (manufactured by Nippon Paper Industries Co., Ltd.). CMC Daicel 1150 (manufactured by Daicel) is preferred.

[0020] The water-dispersible fine cellulose composition is preferably microcrystalline cellulose-carmellose sodium (hereinafter sometimes referred to as MCC-CMCNa). Microcrystalline cellulose-carmellose sodium, sometimes called "microcrystalline cellulose and sodium carboxymethylcellulose," is a mixture of microcrystalline cellulose and carmellose sodium for easy microdispersion. Specifically, it is typically microcrystalline cellulose coated with or attached to carmellose sodium, preferably microcrystalline cellulose coated with carmellose sodium. The content of microcrystalline cellulose in microcrystalline cellulose-carmellose sodium is typically 50% (w / w) to 95% (w / w), preferably 70% (w / w) to 94% (w / w), and more preferably 80% (w / w) to 93% (w / w). The upper and lower limits can be arbitrarily combined as desired, for example, 50% (w / w) to 93% (w / w). The content of carmellose sodium in microcrystalline cellulose carmellose sodium is typically 5% (w / w) to 50% (w / w), preferably 6% (w / w) to 30% (w / w), and more preferably 7% (w / w) to 20% (w / w). The upper and lower limits can be arbitrarily combined as desired, for example, 5% (w / w) to 20% (w / w).

[0021] Examples of crystalline cellulose / carmellose sodium products include, but are not limited to, Ceolus (registered trademark) RC-591NF (manufactured by Asahi Kasei), Avicel (registered trademark) RC-591, RC-581, and BV1518 (manufactured by FMC Biopolymers).

[0022] When the pharmaceutical composition is in a solid or powder form, the amount of water-dispersible fine cellulose composition, particularly crystalline cellulose carmellose sodium, per dosage unit is, for example, 0.03 mg to 60 mg, preferably 0.3 mg to 45 mg, more preferably 0.6 mg to 37.5 mg, even more preferably 0.9 mg to 30 mg, even more preferably 1.5 mg to 22.5 mg, even more preferably 2.25 mg to 18 mg, even more preferably 3 mg to 15 mg, and even more preferably 3 mg to 9 mg. The upper and lower limits can be arbitrarily combined as desired, for example, 0.03 mg to 45 mg.

[0023] The "solvent" used in the pharmaceutical composition of the present invention, or the solvent for dissolving or dispersing the pharmaceutical composition of the present invention, typically includes, but is not limited to, water (purified water, water for injection, or other pharmaceutically acceptable water), a mixture of water and a water-miscible solvent such as an alkanol having 1 to 7 carbon atoms, an aqueous dextrose solution, etc. Water is preferred.

[0024] When the pharmaceutical composition is in the form of a solution, dispersion, suspension, paste, or lotion, the concentration (proportion of total weight) of the water-dispersible fine cellulose composition used in the present invention, in particular microcrystalline cellulose-carmellose sodium, relative to the pharmaceutical composition is, for example, typically 1.7% (w / w) to 9.1% (w / w), preferably 1.9% (w / w) to 9.1% (w / w), more preferably 2.4% (w / w) to 9.1% (w / w), even more preferably 2.9% (w / w) to 9.1% (w / w), even more preferably 2.9% (w / w) to 8.3% (w / w), even more preferably 2.9% (w / w) to 6.5 (w / w), even more preferably 2.9% (w / w) to 5.7 (w / w), and even more preferably 2.9% (w / w) to 4.8 (w / w). The upper and lower limits can be arbitrarily combined as desired, for example, 2.9% (w / w) to 8.3% (w / w).

[0025] As used herein, the term "base" refers to a pharmaceutical additive, or a combination of pharmaceutical additives, among the components of a pharmaceutical composition, that primarily functions to retain and sustainably release a drug in the ear. Specifically, the term "base" refers to, but is not limited to, a water-dispersible fine cellulose composition, a water-dispersible fine cellulose composition and water, or the remaining pharmaceutical additives after removing the drug from the pharmaceutical composition. When the base is in a liquid state, it may also be referred to as a "base liquid."

[0026] As used herein, "gelation" or "gel formation" means that a fluid pharmaceutical composition or base temporarily or continuously loses its fluidity, and "gel-forming ability" means the property of gelling. If a pharmaceutical composition becomes too fluid, the pharmaceutical composition will be expelled from the external auditory canal or Eustachian tube. Therefore, it is preferable that the pharmaceutical composition or base quickly gels temporarily or continuously after intraaural administration. Methods for evaluating the presence or absence of gel formation and gel-forming ability include, but are not limited to, a method in which 300 μL, 200 μL, 100 μL, or 50 μL of the pharmaceutical composition or base is dropped onto a plastic petri dish or test tube, the dish is capped, and the tube is allowed to stand on the water surface of a water bath set at 37°C or on a heat block. 10 seconds, 15 seconds, 20 seconds, 30 seconds, 1 minute, 5 minutes, 10 minutes, 15 minutes, or 30 minutes after the dish or test tube is left standing, the dish or test tube is lifted and immediately inverted, and the behavior of the pharmaceutical composition or base is visually observed for 10 seconds after the dish or test tube is inverted (hereinafter, this method may be referred to as a gel-forming ability test). In the gel-forming ability test, samples left to stand on the water surface of a water bath set at 37°C for 10 seconds, 15 seconds, 20 seconds, 30 seconds, 1 minute, 5 minutes, or 10 minutes typically show no visible "falling" or "sideways flowing" for at least 5 seconds after inversion, and preferably show no visible "falling" or "sideways flowing" for at least 10 seconds after inversion. Note that if the pharmaceutical composition or base is a solid or powder and is to be dissolved or dispersed in an appropriate or specified amount of purified water or other pharmaceutically acceptable solvent before intraaural administration, the fluidity should be evaluated after dissolving or dispersing in an appropriate or specified amount of purified water or other pharmaceutically acceptable solvent (hereinafter, in the method for evaluating a pharmaceutical composition or base described herein, if the pharmaceutical composition or base cannot be appropriately evaluated in its solid or powder state, each item should be evaluated after dissolving or dispersing in an appropriate or specified amount of purified water or other pharmaceutically acceptable solvent).

[0027] Preferably, the pharmaceutical compositions of the present invention do not flow quickly or exhibit poor fluidity after intraaural administration. A method for evaluating the fluidity of the pharmaceutical compositions or bases of the present invention includes, but is not limited to, placing a stainless steel rectangular tray (e.g., a Clover 18-8 shallow rectangular tray) in a chromatography chamber (M-600FN, manufactured by TAITEC) at a set temperature of 37°C so that the angle between the bottom and the horizontal is 30°, gently dropping 0.5 mL of the pharmaceutical composition or base onto the bottom of the rectangular tray, and measuring the distance traveled by the pharmaceutical composition or base down the bottom of the rectangular tray within 60 seconds after dropping (hereinafter, sometimes referred to as a fluidity test). In this fluidity test, the shorter the distance traveled within 60 seconds after dropping, the more likely the pharmaceutical composition is to avoid being discharged from the external auditory canal or Eustachian tube and remain in the ear. The preferred migration distance for 60 seconds after dropping is typically 3.0 cm or less, preferably 2.5 cm or less, more preferably 2.0 cm or less, even more preferably 1.8 cm or less, and even more preferably 1.5 cm or less. The lower limit is 0 cm.

[0028] As used herein, "viscosity" or "static viscosity" refers to a viscosity obtained by dynamic measurement using a viscoelasticity measuring device (hereinafter referred to as a rheometer). The shear rate during measurement is set to a value that allows the sample to be considered stationary. For example, the viscosity can be obtained by a dynamic measurement method (hereinafter sometimes referred to as a static viscosity measurement test) using an Anton Paar rheometer (MCR302, plate: MEASURING CONE CP50-1, hood: H-PTD200, gap: 0.3 mm) under the following measurement conditions: sample volume: 1 mL, shear rate: 0.01 (1 / s), normal force: 0 N, torque reliability range: 1 μN m or more, and temperature: 37°C. Note that if the static viscosity measurement test does not exceed the torque reliability range, it is considered the limit of quantitation (LOQ).

[0029]

[0033] Preferably, the resting viscosity of the pharmaceutical composition of the present invention increases rapidly after intraaural administration using a syringe and needle commonly used in medical settings, so that the composition does not escape from the external auditory canal or Eustachian tube. The method for measuring the resting viscosity is not particularly limited. For example, the viscosity measured 10, 20, or 30 seconds after the start of the static viscosity measurement test, or 10, 20, 30, 60, 90, 120, 150, 180, 210, 240, 270, or 300 seconds after changing the measurement conditions from those of the dynamic viscosity measurement test described below to those of the static viscosity measurement test, is typically 10 Pa·s or higher, preferably 20 Pa·s or higher, more preferably 30 Pa·s or higher, even more preferably 50 Pa·s or higher, and even more preferably 100 Pa·s or higher. Preferably, the viscosity 120 seconds after changing from the measurement conditions of the dynamic viscosity measurement test described below to the measurement conditions of the static viscosity measurement test is typically 10 Pa·s or more, preferably 20 Pa·s or more, more preferably 30 Pa·s or more, even more preferably 50 Pa·s or more, and even more preferably 100 Pa·s or more. More preferably, the viscosity 60 seconds after changing from the measurement conditions of the dynamic viscosity measurement test described below to the measurement conditions of the static viscosity measurement test is typically 10 Pa·s or more, preferably 20 Pa·s or more, more preferably 30 Pa·s or more, even more preferably 50 Pa·s or more, and even more preferably 100 Pa·s or more. Note that if the viscosity is too high, there is a risk that the retained matter will come off the walls of the ear after administration and be expelled from the ear canal or Eustachian tube, and therefore the upper limit of the viscosity is typically 20,000 Pa·s or less.

[0030] As used herein, "dynamic viscosity" refers to a viscosity obtained by dynamic measurement using a rheometer, characterized by setting the shear rate during measurement to a rate at which the sample can be considered to be in a fluid state. For example, dynamic viscosity can be obtained using an Anton Paar rheometer (MCR302, plate: MEASURING CONE CP50-1, hood: H-PTD200, gap: 0.3 mm) under the following measurement conditions: sample volume: 1 mL, shear rate: 1000 (1 / s), normal force: 0 N, torque confidence range: 1 μN m or more, and temperature: 37°C (hereinafter sometimes referred to as a dynamic viscosity measurement test). Note that if the torque confidence range is not exceeded in the dynamic viscosity measurement test, the LOQ is defined as the viscosity.

[0031] The pharmaceutical composition of the present invention can preferably be administered intraaurally using a syringe and needle commonly used in medical settings. The dynamic viscosity of a pharmaceutical composition affects the maximum administration pressure during injection; high dynamic viscosity can hinder easy intraaural administration; therefore, a low dynamic viscosity is preferred. That is, the viscosity of the pharmaceutical composition of the present invention preferably decreases when shear force is applied. The method for measuring dynamic viscosity is not particularly limited. For example, when measuring dynamic viscosity using the dynamic viscosity measurement test, the viscosity of the pharmaceutical composition, base, or base solution is typically less than 10 Pa·s, preferably less than 5 Pa·s, more preferably less than 1 Pa·s, even more preferably less than 0.25 Pa·s, and even more preferably less than 0.20 Pa·s during the dynamic viscosity measurement test. Preferably, 30 seconds after changing the measurement conditions of the dynamic viscosity measurement test, the viscosity is typically less than 10 Pa·s, preferably less than 5 Pa·s, more preferably less than 1 Pa·s, even more preferably less than 0.25 Pa·s, and even more preferably less than 0.20 Pa·s. The lower limit is 0 Pa·s.

[0032] As used herein, "complex viscosity" refers to the viscosity obtained by dynamic measurement using a rheometer. When a thermoreversible gel-forming polymer is used as a base, the viscosity of the base or pharmaceutical composition can be evaluated. Examples of methods for measuring complex viscosity include, but are not limited to, methods using a rheometer. Examples of methods for measuring the complex viscosity of a pharmaceutical composition or base using a rheometer include, but are not limited to, an Anton Paar MCR302 rheometer equipped with a measuring cone CP25-2 plate, a hood H-PTD200, and a 0.3 mm gap, a sample volume of 0.3 mL, and measurement conditions of a shear strain of 5%, an angular frequency of 50 rad / s, a normal force of 0 N, and a torque confidence range of 1 μN·m or greater (hereinafter sometimes referred to as a complex viscosity measurement test). Note that the LOQ is defined as a value that does not exceed the torque confidence range under the complex viscosity measurement test conditions.

[0033] The pharmaceutical composition of the present invention preferably has a low maximum administration pressure (maximum pressurization, maximum pushing pressure) when extruding the pharmaceutical composition so that it can be easily administered into the ear using a syringe and injection needle. A method for measuring the maximum administration pressure includes, but is not limited to, a method using a load cell. For example, a method for measuring the maximum administration pressure using a load cell includes connecting a load cell (LC2-3305B-200N, contact area 25 mm) to a creep meter (RHEONER2 CREEPMETER RE2-33005C, manufactured by YAMADEN). 2、By installing a measurement device (manufactured by YAMADEN), putting an appropriate amount of pharmaceutical composition or base into a Luer lock syringe (SOFT-JECT S5010-LL, manufactured by Henke sass wolf), attaching a 25G injection needle (25G×60mm, manufactured by TOP), removing bubbles and adjusting the filling volume to 0.5 mL to prepare a measurement sample, setting the sample on the measurement device, and measuring the force required to push it at a speed of 1 mm / second in the vertical direction (hereinafter, sometimes referred to as the administration pressure measurement test). However, it is not limited to this. The measurement temperature is not particularly limited as long as it is within the range of room temperature or about 37°C to 38°C. The preferred maximum administration pressure in the administration pressure measurement test is typically 10 N or less, preferably 9 N or less, more preferably 7.2 N or less, still more preferably 6 N or less, and even more preferably 5 N or less. The lower limit of the maximum administration pressure is 0 N.

[0034] In this specification, the "sol-gel transition point" means the temperature at which a pharmaceutical composition changes from a sol state to a gel state. Examples of methods for evaluating the sol-gel transition point include, but are not limited to, methods using a rheometer or the like. In this specification, for example, when measuring the complex viscosity of a pharmaceutical composition or base using a rheometer (MCR302) manufactured by Anton Paar, the storage modulus (G'), which is a solid element, and the loss modulus (G"), which is a liquid element, are simultaneously quantified. When G'<G", it is in the sol state, when G'>G", it is in the gel state, and the temperature at which G' = G" can be defined as the sol-gel transition point.

[0035] In this specification, "slowly releasing a drug" or "sustained release" means that after administering a pharmaceutical composition, the drug continues to elute or has the property of continuing to elute for a certain period. Although there is no particular limitation on the elution rate, since the purpose of slowly releasing a drug is to reduce the number of administrations and / or maintain the drug concentration at the target site constant to sustain the drug effect and reduce side effects, it is preferable that the drug is slowly released.

[0036] A typical method for evaluating sustained release of a drug is, for example, a method based on the method described in "Dose-dependent sustained release of dexamethasone in inner ear cochlear fluids using a novel local delivery approach" (Xiaobo Wang, 2009 Audiol Neurotol 14:393-401). The method involves using a plate well (e.g., Transwell (registered trademark) or Snapwell (trademark) (6 wells, 12 mm diameter inserts, 0.4 μm pore size)) that can separate the upper and lower layers with an insert (insert membrane) with a permeable membrane. 2 mL of the dissolution test solution is placed in a plate well (size, Corning), and the dissolution test solution is heated to 37°C ± 2°C together with the plate well. Then, the insert membrane is fitted into the plate well, and 0.5 mL of a pharmaceutical composition containing one or more drugs is placed on the insert membrane. The plate well is heated to 37°C ± 2°C and shaken at a speed at which the water surface of the dissolution test solution fluctuates (for example, a speed of 100 rpm with a chromato chamber M-600FN manufactured by TAITEC), thereby dissolving the drug from the pharmaceutical composition. For example, the insert membrane is shaken at one of 1, 2, 4, 6, or 24 hours after the start of the dissolution test, or at two or more of 1, 2, 4, 6, or 24 hours after the start of the dissolution test, as necessary. Examples of such methods include, but are not limited to, a method in which the drug concentration in the lower layer of the plate is quantified and the dissolution rate is calculated from the drug concentration (hereinafter sometimes referred to as a dissolution test); or a method in which 2 mL of the pharmaceutical composition is placed into a plate well (e.g., a CELLSTAR™ 6-well plate, SC (manufactured by Greiner)) so that the pharmaceutical composition is spread evenly on the bottom of the well, and 3 mL of elution solution is gently added on top of it, while running it along the wall of the well, to initiate dissolution of the drug from the pharmaceutical composition; the drug concentration in the elution solution is quantified at one time point, for example, 1, 2, 4, 6, or 24 hours after the start of the dissolution test, or at two or more time points, if necessary, at 1, 2, 4, 6, or 24 hours after the start of the dissolution test, and the dissolution rate is calculated from the drug concentration. A dissolution test method is preferred.The method for quantifying the drug may be any analytical method optimal for the quantification of the drug, including, but not limited to, high performance liquid chromatography (HPLC) and a fluorometer. The dissolution test medium is not particularly limited as long as it can appropriately evaluate dissolution. When the solubility of the drug is independent of pH, the dissolution test medium may include, but is not limited to, phosphate buffered saline (hereinafter sometimes referred to as PBS; phosphate buffered salts) such as D-PBS(-). When the solubility of the drug is pH-dependent or the solubility of the drug is low, a dissolution test medium suitable for each drug (e.g., a solvent capable of dissolving 100% of a certain amount of drug) may be used, including, but not limited to, saline or the addition of a low concentration of a surfactant (e.g., polysorbate 20) to a solvent.

[0037] As used herein, the term "sustained drug release" in the dissolution test refers to a drug dissolution rate per appropriate dissolution test time that varies depending on the ear disease and / or drug. For example, this term means that the drug dissolution rate at 1 hour after the start of the dissolution test is typically 40% or less, preferably 35% or less, more preferably 25% or less, and even more preferably 20% or less. It also means that the drug dissolution rate at 2 hours after the start of the dissolution test is typically 50% or less, preferably 45% or less, more preferably 35% or less, and even more preferably 30% or less. It also means that the drug dissolution rate at 4 hours after the start of the dissolution test is typically 70% or less, preferably 65% ​​or less, more preferably 55% or less, and even more preferably 50% or less. It also means that the drug dissolution rate at 6 hours after the start of the dissolution test is typically 80% or less, preferably 75% or less, more preferably 650% or less, and even more preferably 60% or less. Furthermore, for example, it means that the drug dissolution rate 24 hours after the start of the dissolution test is typically 90% or less, preferably 85% or less, more preferably 75% or less, and even more preferably 70% or less.

[0038] As used herein, "retention" or "retention" refers to the property of a drug contained in a pharmaceutical composition administered into the ear to avoid being discharged from the external auditory canal or Eustachian tube and to remain temporarily or continuously at the administration site. Methods for evaluating retention include, but are not limited to, PET (Positron Emission Tomography) imaging, SPECT (Single Photon Emission Computed Tomography) imaging, tests using fluorescent dyes, and MRI (Magnetic Resonance Imaging). As used herein, a test using a fluorescent dye refers to, for example, a test in which a pharmaceutical composition containing a drug encapsulated or labeled with a fluorescent dye is administered into the ear and the amount of fluorescent dye remaining in the ear after a certain period of time is quantified. The retention rate can be calculated from the administered amount and the remaining amount.

[0039] The appropriate drug residual rate in the pharmaceutical composition of the present invention varies depending on the ear disease and / or drug. When the retention of the pharmaceutical composition of the present invention is evaluated by the above-mentioned method, for example, the period after administration until the residual rate reaches 50% or less is typically 6 hours or more, preferably 12 hours or more, more preferably 1 day or more, even more preferably 2 days or more, even more preferably 3 days or more, even more preferably 5 days or more, even more preferably 6 days or more, even more preferably 7 days or more, even more preferably 10 days or more, even more preferably 12 days or more, and even more preferably 14 days or more. Furthermore, the period after administration until the residual rate reaches 20% or less is typically 6 hours or more, preferably 12 hours or more, more preferably 1 day or more, even more preferably 2 days or more, even more preferably 3 days or more, even more preferably 5 days or more, even more preferably 6 days or more, even more preferably 7 days or more, even more preferably 10 days or more, even more preferably 12 days or more, even more preferably 14 days or more, even more preferably 21 days or more, and even more preferably 28 days or more. In the present specification, the term "having retentivity" refers to a case where the retentivity is at least as described above, but is not limited thereto.

[0040] The appropriate frequency of administration of the pharmaceutical composition of the present invention varies depending on the ear disease and / or drug, and is typically twice a day, preferably once a day, more preferably once every two days, even more preferably once every three days, even more preferably once every four days, even more preferably once every five days, even more preferably once every six days, even more preferably once every seven days, even more preferably once every ten days, even more preferably once every 12 days, even more preferably once every 14 days, even more preferably once every 21 days, and even more preferably once every 28 days, but is not limited thereto.

[0041] The dosage of the pharmaceutical composition of the present invention is not particularly limited as long as it can be administered to a patient. Typically, it is an amount that can be administered intraauricularly, preferably an amount necessary to provide useful contact with the tympanic membrane or an amount that can be administered into the tympanic cavity. The dosage is typically 1 μL to 2000 μL, preferably 10 μL to 1500 μL, more preferably 20 μL to 1250 μL, even more preferably 30 μL to 1000 μL, even more preferably 40 μL to 750 μL, even more preferably 50 μL to 600 μL, even more preferably 50 μL to 500 μL, and even more preferably 50 μL to 300 μL. The upper and lower limits can be arbitrarily combined as desired, for example, 1 μL to 1500 μL.

[0042] The pharmaceutical composition of the present invention contains one or more drugs. The "drug" used in the present invention typically includes, but is not limited to, an active ingredient required for the treatment of ear diseases. Preferably, the drug is a low-molecular-weight compound, a nucleic acid, or a protein.

[0043] A low molecular weight compound is a compound having a molecular weight of less than 500, and representative examples of such low molecular weight compounds include, but are not limited to, acetaminophen, diclofenac sodium, and nicardipine hydrochloride. A nucleic acid is a polymeric compound in which nucleotides consisting of a base, a sugar, and phosphate are linked together via phosphodiester bonds. A protein is a polymeric compound formed by the polymerization of L-amino acids in a chain, and examples thereof include, but are not limited to, enzymes, ligands for target receptors, and receptors themselves. A preferred example is a ligand for a target receptor, such as a drug that provides the biological activity of heparin-binding epidermal growth factor.

[0044] Drugs that provide the biological activity of heparin-binding epidermal growth factor used in the present invention include, but are not limited to, HB-EGF (Heparin-Binding Epidermal Growth Factor-like growth factor), human HB-EGF, recombinant human HB-EGF, etc. Typically, HB-EGF is used, preferably human HB-EGF, and more preferably recombinant human HB-EGF.

[0045] Drugs that provide the biological activity of heparin-binding epidermal growth factor for use in the present invention are typically the following proteins: a) a protein comprising an amino acid sequence in which 1 to 10, 1 to 5, 1 to 3, or 1 amino acid is deleted, substituted, inserted, and / or added in the amino acid sequence shown in SEQ ID NO: 1, and having a mitogenic effect and / or a migration-promoting effect on keratinocytes and fibroblasts; b) a protein comprising an amino acid sequence that has an identity of 60% or more, 75% or more, 85% or more, 90% or more, 95% or more, or 100% with the amino acid sequence set forth in SEQ ID NO: 1, and that has a mitogenic and / or migration-promoting effect on keratinocytes and fibroblasts; or c) A protein consisting of the amino acid sequence shown in SEQ ID NO: 1. Preferably, the protein consists of the amino acid sequence shown in SEQ ID NO: 1.

[0046] Drugs that provide the biological activity of the heparin-binding epidermal growth factor used in the present invention can be prepared by those skilled in the art using methods known in the art (e.g., the methods described in WO2014 / 186075) based on the protein sequence information disclosed herein.

[0047] When a drug that provides the biological activity of heparin-binding epidermal growth factor is used, one or a combination of two or more pharmaceutical additives may be added in an appropriate amount within the range in which the drug's efficacy is exerted in order to suppress aggregation of the drug that provides the biological activity of heparin-binding epidermal growth factor or structural changes of the drug that provides the biological activity of heparin-binding epidermal growth factor.

[0048] The pharmaceutical composition of the present invention can be used to treat ear diseases, including, but not limited to, chronic tympanic membrane perforation.

[0049] If desired, one or more pharmaceutical additives may be added to the pharmaceutical composition of the present invention in an appropriate amount within a range that achieves the desired effects of the present invention. Examples of pharmaceutical additives include, but are not limited to, buffers such as PBS, pH adjusters such as dilute hydrochloric acid and sodium hydroxide, isotonicity agents, solubilizers, antioxidants, preservatives, surfactants such as polysorbate 20, etc.

[0050] Examples of solubilizing agents include, but are not limited to, dimethyl sulfoxide (hereinafter sometimes referred to as DMSO), ethanol, dichloromethane, acetone, propylene glycol, polyethylene glycol, glycerol, benzyl alcohol, N,N-dimethylacetamide, etc. DMSO is preferred.

[0051] When a solubilizing agent is added to the pharmaceutical composition of the present invention, the concentration of the solubilizing agent used in the present invention is, for example, typically 0.01% (v / v) to 50% (v / v), preferably 0.1% (v / v) to 30% (v / v), more preferably 1% (v / v) to 10% (v / v), and even more preferably 1% (v / v) to 5% (v / v) relative to the pharmaceutical composition. The upper and lower limits can be arbitrarily combined as desired, for example, 0.01% (v / v) to 30% (v / v).

[0052] The method for producing the pharmaceutical composition of the present invention is described below, but includes known methods including steps such as preparation of the drug, dispersion of the base, pH adjustment, filling, sterilization, lyophilization of the drug and base, final mixing of the drug and base, and terminal sterilization.

[0053] <Drug preparation process> The drug preparation process is not particularly limited in terms of the equipment or means used, as long as it is a method that can dissolve, suspend, or disperse a drug in a typical pharmaceutical manner. Examples include, but are not limited to, methods in which the drug is dissolved, suspended, or dispersed in purified water or a buffer such as PBS as a solvent. In the case of poorly soluble drugs, a solubilizing agent such as DMSO may be added to dissolve the drug, but this is not a limitation. Furthermore, in the case of proteins, etc., a surfactant such as polysorbate 20 may be added to prevent aggregation and / or adsorption to the container, but this is not a limitation.

[0054] <Base dispersion and pH adjustment process> The base dispersion step can be carried out by any method capable of pharmacologic dispersion of the water-dispersible fine cellulose composition serving as the base, particularly crystalline cellulose-carmellose sodium, without particular limitations on the equipment or means used. For example, the base can be added to a solvent such as purified water and dispersed by stirring using a stirrer to obtain a base dispersion, but this method is not limited thereto. Furthermore, the base dispersion can be pulverized using a wet pulverizer to reduce viscosity during administration and improve viscosity after administration, but this method is not limited thereto. For the pH adjustment step, the pH can be adjusted by adding a pH adjuster such as dilute hydrochloric acid or sodium hydroxide, if desired, within a range that achieves the desired effects of the present invention, but this method is not limited thereto.

[0055] <Filling and sterilization process> In the filling and sterilization steps, the equipment and means are not particularly limited as long as they are generally pharmaceutically acceptable for filling and sterilization. For example, a method of dry-heat sterilizing the dispersed base liquid and filling it into a container such as a vial or a device can be mentioned, but is not limited to this. In this specification, sterilization means killing or removing microorganisms present in the pharmaceutical composition, container, device, etc. Examples of sterilization methods include, but are not limited to, dry-heat sterilization, heat sterilization, autoclaving, sterilization with chemicals, radiation sterilization, and filtration sterilization.

[0056] <Lyophilization process of drug and base> The dissolved drug solution and base dispersion can be frozen or lyophilized. In the lyophilization step of the drug, there are no particular limitations on the equipment or means used, as long as it is a method that can be used for pharmaceutical lyophilization.

[0057] <Final mixing process of drug and base> The pharmaceutical composition of the present invention can be provided as, but is not limited to, a frozen dispersion, refrigerated dispersion, or lyophilized product in which the drug and base are mixed, or as a frozen solution, refrigerated solution, or lyophilized product of the drug and base dispersion filled in separate containers. In the case of separate containers, the pharmaceutical composition for intraaural administration of the present invention can be prepared, for example, by adding the base dispersion to a vial containing the drug and mixing them, or by adding the drug or base dispersion from a vial containing the drug or the base dispersion to another empty vial and mixing them, but is not limited to this.

[0058] <Final sterilization process> The pharmaceutical composition for intraotic administration of the present invention may include a terminal sterilization step in its manufacturing process in order to provide a formulation that is free of or safe from infectious microorganisms. Sterilization refers to the killing or removal of microorganisms present in the pharmaceutical composition for intraotic administration, its container, or device. The sterilization method is not particularly limited as long as it is appropriately changed depending on the stability of the drug. For example, typical methods include dry heat sterilization, heat sterilization, autoclaving, sterilization with chemicals, radiation sterilization, and filtration sterilization, and autoclaving is preferred.

[0059] The container for filling the pharmaceutical composition of the present invention can be selected depending on the intended use. As used herein, the term "container" is not particularly limited as long as it can be sealed. Examples include, but are not limited to, vials, ampoules, syringes, bottles, and other large-capacity containers.

[0060] The pharmaceutical composition of the present invention can be delivered to a treatment site using a device. As used herein, the term "device" refers to a medical instrument or apparatus for delivering a pharmaceutical composition to a treatment site, and can be selected depending on the treatment site. Examples of such devices include, but are not limited to, devices in which the pharmaceutical composition is filled into a prescribed dosage form, such as a syringe (including disposable syringes), and is fitted with a needle, cannula, or catheter; double-barrel syringes for delivering two solutions simultaneously or while mixing; sprayers for delivering a spray to a treatment site; and pump-type devices in which the tip of a catheter connected to an osmotic pump is placed near the round window membrane and the osmotic pump is implanted behind the ear, continuously administering a drug solution onto the round window membrane. Preferably, the device is a syringe fitted with a needle. The length and thickness of the injection needle, cannula, or catheter can be selected appropriately depending on the treatment site and the patient. However, when a drug is administered locally to an affected area of ​​the ear or its vicinity by piercing the tympanic membrane, a hole will be created in the tympanic membrane after administration, so it is typically preferable to select an injection needle, cannula, or catheter with a gauge of 25 to 30 G.

[0061] The device can also be used as a container. For example, a syringe can be prefilled with a solution or other liquid to provide a prefilled syringe liquid formulation or an autoinjector liquid formulation. This eliminates the need for dissolving procedures and other operations in the medical field, allowing for more rapid response.

[0062] Examples of materials for the container or device include, but are not limited to, glass, plastic, etc. Furthermore, the container or device may be surface-treated, and may be subjected to, but is not limited to, silica coating, silicon coating, sulfur treatment, various low-alkali treatments, etc.

[0063] The present invention includes a method for retaining a pharmaceutical composition for intra-otic administration containing one or more drugs in the ear by using a water-dispersible fine cellulose composition. The present invention also includes a method for retaining a pharmaceutical composition for intra-otic administration containing one or more drugs in the ear by using microcrystalline cellulose-carmellose sodium.

[0064] The terms "intraaural administration," "for intraaural administration," "water-dispersible fine cellulose composition," "crystalline cellulose-carmellose sodium," "drug," and "retention" used in the method of the present invention can be applied as is to the descriptions of the pharmaceutical composition of the present invention. In a pharmaceutical composition for intra-ear administration containing one or more drugs, the amounts and methods of each component in the method of the present invention for retaining the pharmaceutical composition in the ear using a water-dispersible fine cellulose composition can be directly applied to the pharmaceutical composition of the present invention. [Example]

[0065] The present invention will be described in more detail below with reference to examples, but the present invention should not be construed as being limited thereto.

[0066] In the following examples, crystalline cellulose-carmellose sodium was Ceolus® RC A591NF (MCC content 80% or more, CMCNa content 8.3-13.7%) manufactured by Asahi Kasei, Pluronic® F-127 (hereinafter sometimes referred to as Plu.), a polyoxyethylene-polyoxypropylene triblock copolymer, was manufactured by BASF, sodium hyaluronate was manufactured by ACROS ORGANICS, sodium alginate was I-5 manufactured by Kimika, CMCNa was CMC Daicel 1150 manufactured by Daicel, methylcellulose (MC) was Metolose SM-4000 manufactured by Shin-Etsu Chemical Co., Ltd., hydroxyethyl cellulose (HEC) was SE900 manufactured by Daicel, hydroxypropyl cellulose (HPC) was HPC-H manufactured by Nippon Soda, hydroxypropyl methylcellulose (HPMC) was Metolose 60SH-4000 manufactured by Shin-Etsu Chemical Co., Ltd., polyvinylpyrrolidone was Kollidon 12PF manufactured by BASF, and pectin was GENU HM manufactured by Sansho. Pectin USP-H, Carbopol 971P NF manufactured by Lubrizol, carrageenan GENUVISCO PJ-JPE manufactured by Sansho, karaya gum DSP Gokyo Food & Chemical Karaya Gum Matsu M-150 manufactured by Gokyo Food & Chemical, xanthan gum Xanthan Gum PA manufactured by Ina Foods, DMSO manufactured by Kanto Chemical, polysorbate 20 manufactured by Kanto Chemical, FITC dextran (10kDa) Fluorescein isothiocyanate-dextran, average mol wt 10000 manufactured by Sigma-Aldrich, FITC dextran (70kDa) Fluorescein isothiocyanate-dextran, average mol wt 70000 manufactured by Sigma-Aldrich, FITC dextran (150kDa) Fluorescein isothiocyanate-dextran, average mol wt The reagents used were 150,000 and manufactured by Sigma-Aldrich (dextran will hereafter be referred to as DEX), acetaminophen manufactured by Yamamoto Chemical Industry, diclofenac sodium manufactured by FUJIFILM, nicardipine hydrochloride manufactured by Tokyo Kasei, and saline manufactured by Otsuka Pharmaceutical.Two types of PBS were used in the following examples: 1x PBS (pH 7.4) manufactured by Gibco, and D-PBS(-) manufactured by FUJIFILM. In the examples, when only "%" is displayed for the concentrations of MCC·CMCNa and Plus, this means "% (w / w)."

[0067] Experiment 1: Evaluation of gel-forming ability <Preparation Example 1-1> Preparation of 3.8% MCC·CMCNa base solution 400 mg of MCC·CMCNa was weighed out and dispersed in 10 g of purified water to prepare a 3.8% MCC·CMCNa base solution (Example 1).

[0068] <Preparation Example 1-2> Preparation of 17.0% Plus-PBS base solution 3.4 g of Plu. was weighed out and added to 16.6 g of 1×PBS, and Plu. was dissolved at about 4° C. to prepare a 17.0% Plu.-PBS base solution (Example 2).

[0069] <Test Example 1-1> Gel forming ability test The gel-forming ability at 37°C was evaluated for Example 1 prepared in Preparation Example 1-1 and Example 2 prepared in Preparation Example 1-2. 300 μL of each base liquid was dropped onto a plastic petri dish (1000-035, IWAKI), the dish was covered, and the dish was left to stand on the water surface of a water bath (TR-2A, AS ONE) set at 37°C. After 10, 15, or 20 seconds of standing, each base liquid was removed, and the dish was immediately inverted. The behavior of each base liquid was visually confirmed for 10 seconds after inversion. N=2 samples were tested. A score of × was given for samples that "fell" or "flowed sideways" within 5 seconds, and a score of ◯ was given for samples that "fell" or "flowed sideways" was not observed for at least 5 seconds. The results are shown in Table 1.

[0070] [Table 1]

[0071] The 3.8% MCC·CMCNa (Example 1) tended to form a gel faster than the 17.0% Plus-PBS base solution (Example 2), and it is expected to remain in the ear.

[0072] <Preparation Example 1-3> Preparation of 1.0-9.1% MCC·CMCNa base solution Using the same method as in Preparation Example 1-1, or by diluting the prepared base solution with an appropriate amount of purified water, 1.0%, 1.5%, 1.7%, 1.9%, 2.4%, 2.6%, 2.9%, 3.8%, 4.8%, 5.7%, 6.5%, 7.4%, 8.3%, and 9.1% MCC·CMCNa base solutions (Examples 3 to 16) were prepared. Note that MCC·CMCNa base solutions with concentrations higher than 9.1% could not be prepared because the MCC·CMCNa could not be adequately dispersed in purified water.

[0073] <Test Example 1-2> Gel forming ability test The gel-forming ability at 37°C was evaluated for Examples 3 to 16 prepared in Preparation Example 1-3. The test conditions were the same as Test Example 1-1, except that the standing time was changed to 10 minutes and the drop amount was changed to 300, 200 μL, 100 μL, and / or 50 μL. N=2 samples were tested, and if the sample "fell" or "flowed sideways" within 5 seconds, it was marked with an "X", and if the sample "fell" or "flowed sideways" was not confirmed for at least 5 seconds, it was marked with an "O". If the test was not performed, it was marked with a "-". The results are shown in Table 2.

[0074] [Table 2]

[0075] When a 50μL drop was added, gels were formed with MCC·CMCNa solutions with a concentration of 1.5% or higher. Similarly, gels were formed with MCC·CMCNa solutions with a concentration of 1.9% or higher when a 100μL drop was added, with a concentration of 2.6% or higher when a 200μL drop was added, and with a concentration of 2.6% or higher when a 300μL drop was added.

[0076] Experiment 2: Evaluation of fluidity of base liquid <Test Example 2> Fluidity test of base liquid The fluidity of Example 1 prepared in Preparation Example 1-1 and Example 2 prepared in Preparation Example 1-2 was evaluated. A stainless steel square tray was placed in a chromatography chamber (M-600FN, manufactured by TAITEC) set at a temperature of 37°C so that the angle between the bottom and the horizontal was 30 degrees. 0.5 mL of each base solution was gently dropped onto the bottom of the tray, and the distance traveled by the base solution down the bottom of the tray was measured every 10 seconds for 60 seconds after dropping (N=3). The results are shown in Figure 1. Note that the error bars in Figure 1 indicate standard deviation.

[0077] The 3.8% MCC·CMCNa base solution (Example 1) has a shorter migration distance than the 17.0% Plus-PBS base solution (Example 2), and is therefore expected to allow the drug to remain in the ear for a longer period of time.

[0078] Experiment 3: Evaluation of administration pressure A base liquid having a viscosity above a certain standard was prepared using pharmaceutical additives that impart viscosity, and the maximum administration pressure applied during injection was measured.

[0079] <Preparation Example 3-1> Preparation of each base liquid Sodium hyaluronate, sodium alginate, CMCNa, MC, HEC, HPC, HPMC, polyvinylpyrrolidone, pectin, carbopol, carrageenan, karaya gum, and xanthan gum were used as pharmaceutical additives to impart viscosity. Each pharmaceutical additive was added to purified water to achieve the base concentration shown in Table 3, and dissolved, etc. to prepare the base solutions of Examples 17 to 29. The above-mentioned base concentrations were measured using a rheometer (all rheometers used in this example were MCR302, manufactured by Anton Paar) equipped with a plate (MEASURING CONE CP25-2, manufactured by Anton Paar) and a hood (all hoods used in this example were H-PTD200, manufactured by Anton Paar) with a 0.3 mm gap width, using a 0.3 mL sample volume under the following measurement conditions: shear strain 5%, angular frequency 50 rad / s, normal force 0 N, temperature starting at 25°C and then heating to 37°C at a rate of 1°C / 10 s. Each base concentration was set so as to have a complex viscosity of 1650 mPa s or greater at 37°C. Note that although the complex viscosity of polyvinylpyrrolidone solutions decreases upon heating, 70.0% polyvinylpyrrolidone was used because its complex viscosity at 25°C exceeded 1650 mPa s. A 17.0% Plus-PBS base solution (Example 30) was prepared by adding the base to 1x PBS to the concentration shown in Table 3 and dissolving it by stirring at about 4°C. Purified water (Example 31) was used as a base solution containing no pharmaceutical additives to impart viscosity.

[0080] <Test Example 3-1> Measurement of maximum administration pressure during injection (at 25°C) The maximum administration pressure applied during injection was measured for some of the base solutions prepared in Preparation Example 1-3 (Examples 9 to 11) and each of the base solutions prepared in Preparation Example 3-1 (Examples 17 to 31). The administration pressure was measured using a RHEONER2 CREEPMETER (RE2-33005C, manufactured by YAMADEN) with a load cell (LC2-3305B-200N, contact area 25 mm 2The measurement was performed in a room with a temperature control system (YAMADEN, manufactured by Apiste) and the temperature of the measurement environment was controlled at 25°C using a precision air conditioner (PAU-300S-HC, manufactured by Apiste). 1 mL of each of the base solutions from Examples 9 to 11 or 17 to 30 or purified water (Example 31) was filled into a Luer-lock syringe (SOFT-JECT S5010-LL, manufactured by Henke Sass Wolf). A 25G injection needle (25G x 60 mm, manufactured by Top) was attached, air bubbles were removed, and the filled liquid volume was adjusted to 0.5 mL. The syringe was placed in the measuring device and the injection pressure was measured by pushing it vertically at a speed of 1 mm / sec (N=3). The results are shown in Table 3. Note that in Example 24, the injection pressure load was too high during the measurement, causing the injection needle to come off; therefore, the lowest value of the maximum injection pressures (N=3) was shown instead of the average value. A "-" symbol was used to indicate that the test was not performed.

[0081] [Table 3]

[0082] <Test Example 3-2> Measurement of maximum administration pressure during injection (at 37°C) The maximum administration pressure during injection was measured for some of the base solutions (Examples 3, 6, 9-16) prepared in Preparation Example 1-3. The test conditions were the same as those in Test Example 3-1, except for the temperature conditions. The temperature conditions were simulating the temperature changes that occur during intraauricular administration, and the test was conducted with the entire injection needle connected to the syringe immersed in water at 37°C ± 0.5°C (N = 3). The results are shown in Table 4.

[0083] [Table 4]

[0084] The maximum administration pressure for MCC·CMCNa solutions with concentrations of 1.0 to 6.5% was approximately 5 N, and for the highest concentration, 9.1%, the maximum administration pressure was approximately 10 N, suggesting that intraauricular administration is easy.

[0085] Experiment 4: Evaluation of viscosity during injection <Test Example 4-1> Measurement of static viscosity and dynamic viscosity during injection depending on base concentration The viscosity over time of some of the base solutions (Examples 3-6, 9, 11, 13-16) prepared in the same manner as in Preparation Examples 1-3 was measured under conditions simulating injection administration. A rheometer (MCR302, Anton Paar) equipped with a plate (Measuring Cone CP50-1, Anton Paar), a hood (H-PTD200, Anton Paar), and a 0.3 mm gap was used. The sample volume was 1 mL. The normal force was 0 N, the torque reliability range was 1 μN·m or greater, and the measurement temperature was 37°C. The shear rate was started at 0.01 (1 / s) to simulate a static state. After 30 seconds, the shear rate was changed to 1000 (1 / s), and then changed again to 0.01 (1 / s) after 60 seconds. If the torque reliability range was not exceeded, the value was considered to be below the limit of quantitation (LOQ). The measurements were performed twice and the average values ​​were obtained. The results are shown in Table 5 and Figure 2.

[0086] [Table 5]

[0087] The static viscosity of the MCC·CMCNa solution increased in a concentration-dependent manner, and the higher the concentration, the faster the viscosity returned to normal. At concentrations of 1.7% or higher, the viscosity exceeded 10 Pa·s 120 seconds after the shear rate was returned to 0.01 (1 / s) (180 seconds after the start of measurement). At concentrations of 2.9% or higher, the viscosity exceeded 10 Pa·s 60 seconds after the shear rate was returned to 0.01 (1 / s) (120 seconds after the start of measurement). At concentrations of 7.4% or higher, the viscosity tended to converge to a similar level. At concentrations of 9.1%, the viscosity tended to decrease approximately 120 seconds after the shear rate was returned to 0.01 (1 / s). When the concentration of the MCC·CMCNa base solution was 1.5% or less, the viscosity was below the limit of quantification at 120 seconds (180 seconds after the start of measurement) after the shear rate was returned to 0.01 (1 / s).

[0088] <Preparation Example 4-1> A 3.8% MCC·CMCNa base solution (Example 32) was prepared by dispersing 0.4 g of MCC·CMCNa in 10 g of purified water. A 3.8% MCC·CMCNa base solution (Example 33) was prepared by dispersing 4 g of MCC·CMCNa in 100 g of purified water and grinding the base solution twice at 100 MPa using a wet micronizer (NanoVeita NVL-ED015-D10L-XT110, manufactured by Yoshida Kikai Kogyo Co., Ltd.).

[0089] <Test Example 4-3> Measurement of particle size of dispersed substances in each base liquid The particle size of each base liquid prepared in Preparation Example 4-1 was measured. Approximately 20 μL of each base liquid was added to a measurement cell filled with purified water, and then stirred with a mini-stirrer. The particle size of the dispersion in each base liquid was measured by laser diffraction / scattering measurement using a particle size distribution analyzer (LA-960V2, manufactured by HORIBA). The results are shown in Table 7.

[0090] <Test Example 4-4> Viscosity measurement of each base liquid The effect of crushing MCC·CMCNa on viscosity was investigated. The viscosity change of each base liquid prepared in Preparation Example 4-1 was measured under conditions simulating injection administration. The test conditions were the same as those in Test Example 4-2, except for the number of measurements (N=3). The results are shown in Table 6.

[0091] [Table 6]

[0092] Experiment 5: Effect of DMSO addition on viscosity Assuming that the drug is poorly water-soluble, the viscosity of the base solution was evaluated when DMSO, one of the solubilizing agents, was further added.

[0093] <Preparation Example 5-1> Preparation of MCC·CMCNa base solution (with / without DMSO) 0.33 g, 0.44 g, and 0.55 g of MCC·CMCNa were weighed out and added to 10 mL of purified water, respectively, and stirred to prepare 3.2%, 4.2%, and 5.2% MCC·CMCNa base solutions. Each of the base solutions was mixed with purified water in a 9:1 ratio, or the base solution, purified water, and DMSO were mixed in a 9:0.5:0.5 ratio to prepare 2.9% MCC·CMCNa base solution (Example 34), a 2.9% MCC·CMCNa base solution containing 5.0% (v / v) DMSO (Example 35), a 3.8% MCC·CMCNa base solution (Example 36), a 3.8% MCC·CMCNa base solution containing 5.0% (v / v) DMSO (Example 37), a 4.7% MCC·CMCNa base solution (Example 38), and a 4.7% MCC·CMCNa base solution containing 5.0% (v / v) DMSO (Example 39).

[0094] <Preparation Example 5-2> Preparation of Plus base solution (with / without DMSO) 1.44 g, 1.67 g, and 1.89 g of Plu. were weighed out and added to 8.56 g, 8.33 g, and 8.11 g of 1x PBS, respectively, and stirred and dissolved at approximately 4°C to prepare 14.4%, 16.7%, and 18.9% Plu.-PBS base solutions. Each of the base solutions was mixed with PBS in a 9:1 ratio, or the base solution, 1x PBS, and DMSO were mixed in a 9:0.5:0.5 ratio to prepare 13.0% Plu.-PBS base solution (Example 40), a 13.0% Plu.-PBS base solution containing 5.0% (v / v) DMSO (Example 41), a 15.0% Plu.-PBS base solution (Example 42), a 15.0% Plu.-PBS base solution containing 5.0% (v / v) DMSO (Example 43), a 17.0% Plu.-PBS base solution (Example 44), and a 17.0% Plu.-PBS base solution containing 5.0% (v / v) DMSO (Example 45).

[0095] <Test Example 5-1> Viscosity measurement test of MCC·CMCNa base solution (with / without DMSO) The viscosity change with shear rate was measured for each of the MCC·CMCNa base solutions (Examples 34 to 39) prepared in Preparation Example 5-1. The test conditions were the same as those in Test Example 4-2, except for the number of measurements (N=3) and the measurement time (measurement was continued for 60 seconds after the shear rate was returned to 0.01 (1 / sec)). The results are shown in Table 7 and Figure 3.

[0096] [Table 7]

[0097] <Test Example 5-2> Complex viscosity measurement test of Plu. base solution (with / without DMSO) For each of the Plus base solutions (Examples 40-45) prepared in Preparation Example 5-2, the complex viscosity and sol-gel transition temperature were measured over time with temperature (N=3). Here, complex viscosity refers to the viscosity obtained by dynamic measurement using a rheometer. The sol-gel transition temperature refers to the temperature at which the pharmaceutical composition changes from a sol state to a gel state. The rheometer was equipped with a measuring cone (CP25-2, Anton Paar), a hood, and a 0.3 mm gap. The sample volume was 0.3 mL. The shear strain was 5%, the angular frequency was 50 rad / s, the normal force was 0 N, and the torque reliability range was 1 μN m or greater. The sample temperature was set to 15°C or 20°C at the start of measurement and then increased to 37°C at a rate of 1°C / 10 s. The change in complex viscosity and the sol-gel transition temperature were measured. The results are shown in Table 8 and Figure 4.

[0098] [Table 8]

[0099] The MCC·CMCNa base solution did not show any significant change in viscosity with or without DMSO at any concentration. Furthermore, there was no difference in the rate of viscosity change with shear rate. On the other hand, when comparing the 15.0% Plu.-PBS base solution with the 15.0% Plu.-PBS base solution containing 5.0% (v / v) DMSO, the temperature at which the complex viscosity reached 10,000 mPa·s and the sol-gel transition temperature were shifted 2–3°C lower by the addition of DMSO. This was also the case for the 17.0% Plu.-PBS base solution. The 13.0% Plu.-PBS base solution, regardless of the presence or absence of DMSO, did not show an increase in complex viscosity or sol-gel transition with increasing temperature. These results demonstrate that the viscosity of the MCC·CMCNa base solution is not affected by DMSO.

[0100] Experiment 6: Evaluation of sustained drug release The sustained release of drug-containing liquid formulations based on MCC·CMCNa was evaluated using several model drugs. The model drugs used were FITC-dextran (10 kDa, 70 kDa, 150 kDa) (hereinafter referred to as FITC-DEX) as a protein drug model, acetaminophen and diclofenac sodium as acidic small molecule drug models with different partition coefficients, and nicardipine hydrochloride as a basic small molecule drug model.

[0101] <Preparation Example 6-1> Preparation of FITC-DEX (10 kDa)-containing liquid 50 mg of FITC-DEX (10 kDa) was dissolved in D-PBS(-) to make 5 mL of 1.0% (w / v) FITC-DEX. 200 μL of 1.0% (w / v) FITC-DEX was mixed with 1800 μL of D-PBS(-) to prepare a PBS solution containing 0.1% FITC-DEX (Example 46).

[0102] 945 mg of Plu. was weighed out and added to 4.055 g of 1x PBS, and the Plu. was dissolved at approximately 4°C to prepare an 18.9% Plu.-PBS base solution. 200 μL of 1.0% (w / v) FITC-DEX was mixed with 1800 μL of the base solution to prepare a 17.0% Plu.-PBS solution containing 0.1% FITC-DEX (Example 47).

[0103] A 1.1% MCC·CMCNa solution was prepared by dispersing 0.11 g of MCC·CMCNa in 10 g of purified water. 200 μL of 1.0% (w / v) FITC-DEX was added to 1800 μL of the solution to prepare a 1.0% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 48). Similarly, 200 μL of 1.0% (w / v) FITC-DEX was mixed with 1800 μL each of 1.6% MCC·CMCNa vehicle solution, 1.9% MCC·CMCNa vehicle solution, 2.2% MCC·CMCNa vehicle solution, 3.2% MCC·CMCNa vehicle solution, 4.2% MCC·CMCNa vehicle solution, and 5.2% MCC·CMCNa vehicle solution to prepare 1.5% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 49), 0.1% FITC-DEX, and 1.5% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 49). A 1.7% MCC·CMCNa solution containing ITC-DEX (Example 50), a 1.9% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 51), a 2.9% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 52), a 3.8% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 53), and a 4.7% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 54) were prepared.

[0104] <Test Example 6-1> Dissolution test of liquid preparation containing FITC-DEX (10 kDa) Dissolution tests were performed on each of the liquid formulations (Examples 46 to 54) prepared in Preparation Example 6-1 (N=2). The dissolution tests were performed by placing 2 mL of D-PBS(-) in a plate well of a Snapwell™ (6-well, 12 mm diameter inserts, 0.4 μm pore size, Corning) and heating the plate well in a chromatographic chamber (M-600FN, TAITEC) set to 37°C. The insert membrane was then inserted into the well, and 0.5 mL of each liquid formulation was dispensed onto the insert membrane. The plate was then shaken at 37°C ± 2°C and 100 rpm. At 1, 2, 4, 6, and 24 hours after the start of the test, 0.5 mL of the lower layer of the insert membrane was sampled, and 0.5 mL of D-PBS(-) was added to the upper layer of the plate well. The FITC concentration of the sample solution was measured using a fluorescent plate reader (SpectraMax i3x, manufactured by MOLECULAR DEVICES) at an excitation wavelength of 494 nm and a measurement wavelength of 522 nm, and the dissolution rate was calculated. The results are shown in Table 9.

[0105] [Table 9]

[0106] <Preparation Example 6-2> Preparation of FITC-DEX (70 kDa)-containing liquid 50 mg of FITC-DEX (70 kDa) was dissolved in D-PBS(-) solution to make 5 mL, and 1.0% (w / v) FITC-DEX was obtained. Each liquid preparation (Examples 55 and 56) listed in Table 10 was prepared in the same manner as in Preparation Example 6-1.

[0107] A 3.3% MCC·CMCNa solution was prepared by dispersing 9.9 g of MCC·CMCNa in 290.1 ​​g of purified water. 200 μL of 1.0% (w / v) FITC-DEX was added to 1800 μL of the solution to prepare a 3.0% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 57). Similarly, 200 μL of 1.0% (w / v) FITC-DEX was added to 1800 μL each of 4.4% MCC·CMCNa solution and 5.5% MCC·CMCNa solution to prepare a 4.0% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 58) and a 5.0% MCC·CMCNa solution containing 0.1% FITC-DEX (Example 59).

[0108] <Test Example 6-2> Dissolution test of liquid preparation containing FITC-DEX (70 kDa) For Examples 55 to 59 prepared in Preparation Example 6-2, an FITC-DEX (70 kDa) dissolution test was performed (N=2) under the same test conditions as in Test Example 6-1 (except that the fluorescent plate reader was changed to Infinite M1000PRO, manufactured by TECAN). The results are shown in Table 10.

[0109] [Table 10]

[0110] <Preparation Example 6-3> Preparation of FITC-DEX (150 kDa)-containing liquid 50 mg of FITC-DEX (150 kDa) was dissolved in D-PBS(-) solution to make 5 mL, and 1.0% (w / v) FITC-DEX was obtained. Each liquid preparation (Examples 60 to 64) listed in Table 11 was prepared in the same manner as in Preparation Example 6-2.

[0111] <Test Example 6-3> Dissolution test of FITC-DEX (150 kDa)-containing liquid For Examples 60 to 64 prepared in Preparation Example 6-3, an FITC-DEX (150 kDa) dissolution test was carried out (N=2) under the same test conditions and method as in Test Example 6-2. The results are shown in Table 11.

[0112] [Table 11]

[0113] <Preparation Example 6-4> Preparation of acetaminophen-containing liquid 100 mg of acetaminophen was weighed out and dissolved in purified water to make 10 mL to give 1.0% (w / v) acetaminophen. 120 μL of 1.0% (w / v) acetaminophen was mixed with 1080 μL of 1×PBS to prepare a PBS solution containing 0.1% acetaminophen (Example 65).

[0114] 100 mg of acetaminophen was weighed out and dissolved in 0.05% polysorbate 20 / PBS solution to make 10 mL, creating a 1.0% (w / v) acetaminophen / PBS solution. 18.9 g of Plu. was weighed out and added to 81.1 g of 1x PBS. The Plu. was dissolved at approximately 4°C to prepare an 18.9% Plu.-PBS base solution. 300 μL of the 1.0% (w / v) acetaminophen / PBS solution was mixed with 2700 μL of the base solution to prepare a 17.0% Plu.-PBS solution containing 0.1% acetaminophen (Example 66).

[0115] A 4.2% MCC·CMCNa solution was prepared by dispersing 2.2 g of MCC·CMCNa in 50 mL of purified water. 150 μL of 1.0% (w / v) acetaminophen was added to 1350 μL of the solution to prepare a 3.8% MCC·CMCNa solution containing 0.1% acetaminophen (Example 67).

[0116] <Test Example 6-4> Dissolution test of acetaminophen-containing liquid preparations For Examples 65 to 67 prepared in Preparation Example 6-4, a dissolution test was conducted (N=2) under the same test conditions as in Test Example 6-1 (except that the dissolution test solution was changed to 1xPBS). Measurements were performed using HPLC (ESPRIMO D583 / N, manufactured by Waters) to measure the acetaminophen concentration of the sample solution at a measurement wavelength of 254 nm. The results are shown in Table 12.

[0117] [Table 12]

[0118] <Preparation Example 6-5> Preparation of diclofenac sodium-containing liquid 50 mg of diclofenac sodium was dissolved in D-PBS(-) solution to make 5 mL, and 1.0% (w / v) diclofenac sodium was obtained. Each liquid preparation (Examples 68 to 70) listed in Table 13 was prepared in the same manner as in Preparation Example 6-1.

[0119] <Test Example 6-5> Dissolution test of diclofenac sodium-containing liquid preparation For Examples 68 to 70 prepared in Preparation Example 6-5, a dissolution test was carried out under the same test conditions as in Test Example 6-1. The diclofenac sodium concentration of the sample solution was measured using HPLC (Waters Alliance e2695, manufactured by Waters) at a measurement wavelength of 240 nm. The results are shown in Table 13.

[0120] [Table 13]

[0121] <Preparation Example 6-6> Preparation of a liquid containing nicardipine hydrochloride 25 mg of nicardipine hydrochloride was dissolved in purified water to make 5 mL of solution to prepare 0.5% (w / v) nicardipine hydrochloride. 200 μL of 0.5% (w / v) nicardipine hydrochloride was mixed with 1800 μL of physiological saline to prepare 0.05% nicardipine hydrochloride-containing physiological saline (Example 71).

[0122] 1800 μL of 18.9% Plus-PBS base solution prepared in the same manner as in Preparation Example 6-1 was mixed with 200 μL of 0.5% (w / v) nicardipine hydrochloride to prepare a 17.0% Plus-PBS solution containing 0.05% nicardipine hydrochloride (Example 72).

[0123] 200 μL of 0.5% (w / v) nicardipine hydrochloride was mixed with 1800 μL of 4.4% MCC·CMCNa base solution prepared in the same manner as in Preparation Example 6-1 to prepare a 4.0% MCC·CMCNa solution containing 0.05% nicardipine hydrochloride (Example 73).

[0124] <Test Example 6-6> Dissolution test of liquid preparations containing nicardipine hydrochloride For Examples 71 to 73, dissolution tests were carried out under the same test conditions as in Test Example 6-1, except that the dissolution test solution was physiological saline instead of D-PBS(-). Measurements were carried out using HPLC (Waters Alliance e2695, manufactured by Waters) to measure the nicardipine hydrochloride concentration in the sample solution at a measurement wavelength of 254 nm. The results are shown in Table 14.

[0125] [Table 14]

[0126] Experiment 7: Evaluation of intra-ear retention using fluorescent dyes The intraear retention of a pharmaceutical composition containing MCC·CMCNa was evaluated using FITC-DEX (10 kDa) as a model drug.

[0127] <Preparation Example 7-1> Preparation of FITC-DEX (10 kDa)-containing liquid 70 mg of FITC-DEX (10 kDa) was dissolved in 1×PBS and the resulting solution was adjusted to 5 mL to prepare a 1.4% (w / v) FITC-DEX solution. 400 μL of this solution was mixed with 3600 μL of PBS to prepare a 0.14% (w / v) FITC-DEX-containing PBS solution (Example 74).

[0128] An 18.9% Plu.-PBS base solution was prepared by weighing out 1.89 g of Plu. and adding it to 8.11 g of 1x PBS and dissolving the Plu. at approximately 4°C. 400 μL of the 1.4% (w / v) FITC-DEX solution was mixed with 3600 μL of the 18.9% Plu.-PBS solution to prepare a 17.0% (w / v) Plu. solution containing 0.14% (w / v) FITC-DEX (Example 75).

[0129] A 1.1% MCC·CMCNa solution was prepared by dispersing 0.11 g of MCC·CMCNa in 10 g of purified water. 3600 μL of this solution was mixed with 400 μL of a 1.4% (w / v) FITC-DEX solution to prepare a 1.0% MCC·CMCNa solution containing 0.14% (w / v) FITC-DEX (Example 76). Similarly, 400 μL of 1.4% (w / v) FITC-DEX was mixed with 3600 μL each of 2.2% MCC·CMCNa base solution, 3.2% MCC·CMCNa base solution, and 4.2% MCC·CMCNa base solution to prepare 1.9% MCC·CMCNa solution containing 0.14% (w / v) FITC-DEX (Example 77), 2.9% MCC·CMCNa solution containing 0.14% (w / v) FITC-DEX (Example 78), and 3.8% MCC·CMCNa solution containing 0.14% (w / v) FITC-DEX (Example 79).

[0130] <Test Example 7-2> Retention test of FITC-DEX (10 kDa)-containing liquid A rat ear retention test was conducted for each of the solutions prepared in Preparation Example 7-1 (Examples 74 to 79) (N = 3 to 5). Male Slc:SD rats were purchased from Japan SLC and 6-week-old rats were used in the experiment. Under isoflurane (Pfizer) inhalation anesthesia, a hole was made in the tympanic membrane of the right ear of each rat using a 25G needle (NN-2525R, Terumo). 25 μL or 50 μL of each solution was administered through the hole using a microsyringe (1710LT, Hamilton) equipped with a blunted 25G needle. After administration, the rats were left in a lateral position for approximately 10 minutes before awakening. One or three days later, the rats were exsanguinated and decapitated under anesthesia, and the middle ear cavity was sampled. FITC-DEX was collected from the collected middle ear cavities, and the amount of FITC-DEX remaining in the ears was measured using a fluorescent plate reader (Infinite M1000PRO, manufactured by TECAN). The results are shown in Table 15.

[0131] [Table 15]

[0132] Experiment 8: Evaluation of the efficacy of HB-EGF-containing liquid using a chronic tympanic membrane perforation model A mouse model of chronic tympanic membrane perforation was prepared, and the efficacy of each liquid preparation containing mouse HB-EGF on tympanic membrane perforation was evaluated.

[0133] <Test Example 8-1> Creation of a chronic tympanic membrane perforation model mouse Male CBA / CaJ mice were bred and maintained at the Astellas Pharma Animal Research Facility from broodstock purchased from Charles River Japan. Six-week-old mice were used for the experiments. Under isoflurane (Pfizer) inhalation anesthesia, the left and right tympanic membranes of each mouse were fully perforated using a Rosen probe (52-147-50, Daiichi Medical). Gelfoam® (Pfizer) cut to fit inside the mouse's ear was soaked in a 10 mmol / L KB-R7785 solution (synthesized in-house) as a perforation agent (Hirayama et al., Bioorg Med Chem. 1997, Apr;5(4)765-778) (Morimoro et al., Life Sci. 1997;61(8):795-803) and placed at the perforation site. From the day after perforation creation, the inside of the ear was observed once a day. If the Gelfoam® had disappeared, a new Gelfoam® soaked in perforation agent was placed. If the Gelfoam® had not disappeared, additional perforation agent was administered and allowed to soak into the Gelfoam®. The perforation agent treatment was carried out for 7 days, and after 3 months, the perforation was confirmed under a microscope. Tympanic membranes that had healed naturally were excluded from further testing.

[0134] <Preparation Example 8-1> Preparation of mouse HB-EGF solution Mouse HB-EGF (cyt-068, manufactured by Prospec) was used. 50 μg of HB-EGF was dissolved in 850 μL of distilled water to prepare a 58.824 μg / mL mouse HB-EGF solution. 70 μL of distilled water was added to 400 μL of the 58.824 μg / mL mouse HB-EGF solution to prepare a 50 μg / mL mouse HB-EGF solution.

[0135] <Preparation Example 8-2> Preparation of saline solution (with / without mouse HB-EGF) The 50 μg / mL mouse HB-EGF solution prepared in Preparation Example 8-1 or distilled water was diluted 10-fold with physiological saline to prepare a 5 μg / mL mouse HB-EGF solution (with mouse HB-EGF) (Example 80) or a distilled water / physiological saline solution (without mouse HB-EGF) (Example 81). These solutions were stored at 4°C.

[0136] <Preparation Example 8-3> Preparation of 3.8% MCC·CMCNa solution (with / without mouse HB-EGF) A 4.2% MCC·CMCNa solution was prepared by dispersing 2.2 g of MCC·CMCNa in 50 g of purified water. A 3.8% MCC·CMCNa solution (with mouse HB-EGF) (Example 82) or a 3.8% MCC·CMCNa solution (without mouse HB-EGF) (Example 83) was prepared by mixing the 50 μg / mL mouse HB-EGF solution prepared in Preparation Example 8-1 or distilled water with the 4.2% MCC·CMCNa solution in a 1:9 ratio. These solutions were stored at 4°C.

[0137] <Test Example 8-2> Treatment of chronic tympanic membrane perforation 5 μL of each of the solutions (Examples 80 to 83) prepared in Preparation Examples 8-2 and 8-3 was administered onto the tympanic membrane using a Microman E (M10E, manufactured by Gilson) in mice with chronic tympanic membrane perforation prepared in Test Example 8-1 (N=10 to 12). One month after administration, the surrounding tissue, including the tympanic membrane, was sampled, and the presence or absence of perforation was confirmed after removing any residue. The tympanic membrane perforation healing rate was calculated from the number of perforated tympanic membranes and the number of completely healed tympanic membranes. The results are shown in Table 16.

[0138] [Table 16]

[0139] The healing rate for tympanic membrane perforations using saline solution (with mouse HB-EGF) (Example 80) was 30%, while the healing rate for tympanic membrane perforations using 3.8% MCC·CMCNa solution (with mouse HB-EGF) (Example 82) was 64%. [Industrial Applicability]

[0140] According to the present invention, it is possible to provide a pharmaceutical composition for intraaural administration that can be easily administered intraaurally and has the function of retaining and sustained-release of a drug in the ear.

[0141] While the present invention has been described with reference to specific embodiments, variations and modifications that are obvious to those skilled in the art are within the scope of the present invention.

Claims

1. A pharmaceutical composition for intraaural administration, comprising one or more drugs and a water-dispersible fine cellulose composition which is crystalline cellulose / carmellose sodium.

2. The pharmaceutical composition of claim 1 , further comprising a solvent.

3. The pharmaceutical composition of claim 2 , wherein the solvent is water.

4. 4. The pharmaceutical composition according to claim 2, wherein the proportion of the total weight of the water-dispersible fine cellulose composition is 1.7% (w / w) to 9.1% (w / w) based on the total weight of the pharmaceutical composition.

5. 4. The pharmaceutical composition according to claim 2, wherein the proportion of the total weight of the water-dispersible fine cellulose composition is 2.9% (w / w) to 9.1% (w / w) based on the total weight of the pharmaceutical composition.

6. The pharmaceutical composition according to any one of claims 1 to 5, which sustains the release of a drug when an insert with a permeable membrane is fitted into a plate well containing 2 mL of phosphate buffered saline at 37°C ± 2°C so as to separate the phosphate buffered saline into an upper layer and a lower layer, 0.5 mL of the pharmaceutical composition according to any one of claims 1 to 5 is placed on the insert, and the plate well is shaken at a speed that causes the water surface of the phosphate buffered saline to fluctuate, thereby dissolving the drug.

7. The pharmaceutical composition according to any one of claims 1 to 6, further comprising a solubilizing agent.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the drug is one or more selected from the group consisting of low molecular weight compounds, nucleic acids, and proteins.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the drug comprises a protein consisting of the amino acid sequence shown in SEQ ID NO:1.

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