WO2017142193A1

WO2017142193A1 - Ophthalmic composition having aqueous solution form comprising cyclosporin and hyaluronic acid or salt thereof

Info

Publication number: WO2017142193A1
Application number: PCT/KR2017/000034
Authority: WO
Inventors: 임광진; 정상욱; 박상준; 양호한; 황성주; 강한; 김택선; 이지연
Original assignee: 삼천당제약주식회사
Priority date: 2016-02-15
Filing date: 2017-01-03
Publication date: 2017-08-24
Also published as: KR101635915B1

Abstract

The present invention provides an ophthalmic composition in the form of an aqueous solution comprising cyclosporin and hyaluronic acid or a salt thereof, wherein the ophthalmic composition contains d-α-tocopherol polyethylene glycol succinate as a stabilizer and has excellent physical and chemical stability.

Description

Ophthalmic composition in the form of an aqueous solution comprising cyclosporin and hyaluronic acid or salts thereof

The present invention relates to an ophthalmic composition in the form of an aqueous solution comprising cyclosporin and hyaluronic acid or a salt thereof. More particularly, the present invention relates to ophthalmic compositions having good physical and chemical stability, including d-α-tocopherol polyethylene glycol succinate as stabilizer.

The most common method of dry eye treatment is to temporarily relieve symptoms by administering artificial tears to the eye whenever the symptoms of dry eye appear. Artificial tears contain viscoelastic substances such as carboxymethyl cellulose, chondroitin sulfate, and sodium hyaluronate, which replenish moisture and stabilize the tear film. Sodium hyaluronate exhibits excellent water retention by retaining water molecules in the molecule, thereby relieving dryness of the corneal conjunctiva, which is the external eye tissue of the eye, and healing epithelial cells of the cornea. However, artificial tears containing these substances have limited therapeutic effects because they differ physically and physiologically from natural tears.

Unlike artificial tears that temporarily relieve dry eye, cyclosporin reduces the number of T-lymphocytes in the conjunctival tissue, inhibits cytokine production, regulates the inflammatory response of the ocular surface and lacrimal glands, and increases the number of conjunctival goblet cells. It is known to promote tear production. Currently, Restasis ^™ (a cyclosporine ophthalmic emulsion 0.05%) by Allergan, USA is marketed as a cyclosporine-containing ophthalmic composition for the treatment of dry eye. The formulation contains castor oil as the solubilizer of cyclosporin and is in the form of an emulsion obtained using a crosslinked polyacrylate emulsifier and a surfactant such as polysorbate 80 to emulsify the oil component in an aqueous solution. 368,181). An emulsion refers to a liquid-liquid dispersion system in which at least one liquid phase is dispersed in one liquid phase, and generally has a size distribution of 0.1 μm to several tens of μm. Such emulsions are thermodynamically unstable and have the property of separating into various forms such as coagulation, sedimentation, creaming, grain growth, and coalescence. Commercially available restissis has oily eye irritation, emulsion particles are uneven and sufficient stability is not secured before use.

Recently, studies have been developed on the combination of cyclosporin for the treatment of dry eye due to tearing and sodium hyaluronate, which has a healing effect on corneal epithelial disorder caused by dry eye. For example, US Pat. No. 6,677,304 discloses a topical ophthalmic composition in the form of an aqueous solution comprising cyclosporin and sodium hyaluronate and comprising polysorbate 80 as a stabilizer. According to the patent, it is disclosed that it is stable without deposit formation at room temperature for 12 months.

As a result of evaluating the stability of the ophthalmic composition disclosed in US Pat. No. 6,677,304, the inventors found that the stability of the cyclosporin was drastically reduced to satisfy the Ministry of Food and Drug Safety approval criteria (see Test Example 2 below). ). In addition, the ophthalmic composition disclosed in U.S. Patent No. 6,677,304 was found to produce a precipitate in one week when stored under accelerated storage conditions (40 ± 2 ° C./25% relative humidity or less) (Test Example 1 below). Reference).

The inventors have conducted various studies to improve the physical and chemical stability of ophthalmic compositions in the form of aqueous solutions containing cyclosporin and hyaluronic acid or salts thereof. As a result, it has been found that ophthalmic preparations in the form of aqueous solutions containing cyclosporin and hyaluronic acid or salts thereof have excellent physical and chemical stability, using certain additives, d-α-tocopherol polyethylene glycol succinate, as stabilizers. .

Accordingly, it is an object of the present invention to provide an ophthalmic composition in the form of an aqueous solution comprising cyclosporin and hyaluronic acid or salts thereof comprising d-α-tocopherol polyethylene glycol succinate as a stabilizer.

According to one aspect of the present invention, an ophthalmic composition in the form of an aqueous solution comprising cyclosporin and hyaluronic acid or a salt thereof, wherein the ophthalmic composition comprises d-α-tocopherol polyethylene glycol succinate as a stabilizer. Is provided.

The ophthalmic composition of the present invention may comprise d-α-tocopherol polyethylene glycol succinate at a concentration of 0.01 to 5.00 w / v%, preferably 0.10 to 1.00 w / v%.

In addition, the ophthalmic composition of the present invention may further comprise at least one stabilizing aid selected from the group consisting of polyoxyl 35 castor oil, polysorbate 80, and polyethylene glycol 200. Stabilizing aids such as polyoxyl 35 castor oil, polysorbate 80, polyethylene glycol 200, etc. may be included independently at a concentration of 0.10 to 5.00 w / v%.

According to the present invention, when the d-α-tocopherol polyethylene glycol succinate is contained as a stabilizer, the ophthalmic preparation in the form of an aqueous solution containing cyclosporin and hyaluronic acid or a salt thereof has excellent physical and chemical stability, and thus, It has been found that they can meet the permit criteria. In addition, the ophthalmic composition of the present invention can be sterilized using a conventional antibacterial filter by having an aqueous solution form. Therefore, the ophthalmic composition according to the present invention can be usefully used for the treatment of dry eye and the like.

1 shows evaluation items of the National Eye Institute (NEI) grading system.

Figure 2 shows the results of evaluating the effect of dry eye syndrome according to the National Eye Institute (NEI) grading system through corneal staining using a fluorescent dye (fluorescein). * p <0.05, ** p <0.01 (vs. cyclosporin A alone formulation, # p <0.05 (vs. sodium hyaluronate alone formulation)

The present invention provides an ophthalmic composition comprising an d-α-tocopherol polyethylene glycol succinate as a stabilizer in an ophthalmic composition in the form of an aqueous solution comprising cyclosporin and hyaluronic acid or a salt thereof.

Cyclosporin and hyaluronic acid or salts thereof contained as an active ingredient for treating dry eye in the ophthalmic composition of the present invention may be used in therapeutically effective amounts, respectively. For example, the cyclosporin includes various cyclosporine analogs, such as cyclosporin A, B, C, D, G, and may preferably be cyclosporin A. Cyclosporin, such as cyclosporin A may be present in an aqueous solution of, for example, 0.01 to 2.00 w / v%, preferably 0.01 to 0.20 w / v%, more preferably about 0.05 w / v%, but is not limited thereto. It doesn't happen. The hyaluronic acid or a salt thereof (for example sodium hyaluronate) is, for example, in an aqueous solution of 0.01 to 2.00 w / v%, preferably 0.10 to 0.50 w / v%, more preferably about 0.30 w / v% It may be present in a concentration of, but is not limited thereto.

D-α-Tocopherol polyethylene glycol succinate used as a stabilizer in the pharmaceutical composition of the present invention is' d-α-Tocopherol polyethylene glycol 1000 succinate (d-α-Tocopherol polyethylene glycol) 1000 succinate) ',' vitamin E TPGS ',' TPGS 'and the like. The d-α-tocopherol polyethylene glycol succinate may be used in an amount to maintain the stability of the ophthalmic composition in the form of an aqueous solution, for example, 0.01 to 5.00 w / v% in an aqueous solution, preferably 0.10 to 1.00 w / v%, more preferably at a concentration of about 0.50 w / v%.

In one embodiment, the present invention provides an ophthalmic composition comprising 0.05 w / v% cyclosporin, 0.30 w / v% hyaluronic acid or a salt thereof, and 0.50 w / v% d-α-tocopherol polyethylene glycol succinate. .

The pharmaceutical compositions of the invention may include a stabilizing aid, such as polyoxyl 35 castor oil (polyoxyl 35 castor oil) [for example, "Crescent mode pore ^TM ELP (Cremophor ^TM ELP)"], polysorbate 80, and at least one stabilizing aid selected from the group consisting of polyethylene glycol 200. Stabilizing aids such as polyoxyl 35 castor oil, polysorbate 80, polyethylene glycol 200, and the like may be independently included, for example, at a concentration of 0.10 to 5.00 w / v%.

In one embodiment, the present invention provides 0.05 w / v% of cyclosporin, 0.30 w / v% of hyaluronic acid or a salt thereof, 0.50 w / v% of d-α-tocopherol polyethylene glycol succinate, 1.00 w / of polyoxyl 35 castor oil. An ophthalmic composition comprising v%, and polysorbate 80 0.50 w / v%. In another embodiment, the invention provides 0.05 w / v% of cyclosporin, 0.30 w / v% of hyaluronic acid or a salt thereof, 0.50 w / v% of d-α-tocopherol polyethylene glycol succinate, and 2.00 w of polyoxyl 35 castor oil. Provided is an ophthalmic composition comprising / v%. In another embodiment, the present invention provides 0.05 w / v% of cyclosporin, 0.30 w / v% of hyaluronic acid or a salt thereof, 0.50 w / v% of d-α-tocopherol polyethylene glycol succinate, 2.00 w of polyoxyl 35 castor oil. / v%, and polyethylene glycol 200 1.00 w / v%.

Ophthalmic compositions in the form of aqueous solutions of the present invention may include additives commonly used in the field of ophthalmic compositions. Examples of the additives include pH adjusting agents such as monosodium phosphate and disodium phosphate; Isotonic agents, such as glycerin, sorbitol, mannitol and the like. The pH adjusting agent is used to maintain the pH of the resulting ophthalmic composition is 7.0 to 7.5, it may be added in the form of a phosphate buffer.

The ophthalmic composition of this invention can be sterilized using a conventional disinfection filter (for example, a 0.22 micrometer filter). Since the ophthalmic composition of the present invention has an aqueous solution form, unlike an emulsion preparation containing an oil component, sterilization using a conventional antibacterial filter is possible.

Hereinafter, the present invention will be described in more detail through Examples and Test Examples. However, these Examples and Test Examples are for illustrating the present invention, and the present invention is not limited to these Examples and Test Examples.

실시예Example 1∼25. 비타민 E 1-25. Vitamin E TPGS를TPGS 안정화제로서 함유하는 안과용 조성물의 제조 Preparation of ophthalmic composition containing as stabilizer

According to the ingredients and contents of Tables 1 to 3, containing vitamin E TPGS as a stabilizer or containing vitamin E TPGS as a stabilizer and containing Cremophor ELP, polysorbate 80, and PEG 200 as stabilizing aids An ophthalmic composition was prepared. The content of each component shown in Tables 1 to 3 below represents w / v% unless otherwise indicated.

Completely dissolve cyclosporin A or cyclosporin A and a stabilizing aid in vitamin E TPGS (solution-A). Sodium hyaluronate, monosodium phosphate, disodium phosphate, and glycerin were added to sterile purified water to obtain a solution (solution-B). After solution-A was sufficiently mixed with Solution-B, sterile purified water was added to adjust the total volume to 100 mL, and then filtered through a 0.22 μm filter to prepare an ophthalmic composition. The pH of the obtained ophthalmic composition is as described in following Tables 1-3.

TABLE 1

Ophthalmic Compositions Containing Stabilizers

TABLE 2

Ophthalmic composition containing stabilizer and stabilizing aid

TABLE 3

Ophthalmic composition containing stabilizer and stabilizing aid

비교예Comparative example 1∼10. 1 to 10. 폴리소르베이트Polysorbate 80을 안정화제로서 함유하는 안과용 조성물의 제조 Preparation of an ophthalmic composition containing 80 as a stabilizer

An ophthalmic composition was prepared containing the polysorbate 80 disclosed in US Pat. No. 6,677,304 as a stabilizer with the ingredients and contents of Table 4 below. In addition, ophthalmic compositions containing 0.05 w / v% and 0.30 w / v% of cyclosporin A and sodium hyaluronate, respectively, were prepared using the ingredients and contents shown in Table 5 below. In Table 4 and Table 5 above The content of each component described represents w / v% unless otherwise indicated.

Cyclosporin A was completely dissolved in polysorbate 80 (solution-A). Sodium hyaluronate, sodium hydrogen phosphate 12-hydrate, and sorbitol were added to sterile purified water to obtain a solution (solution-B). After solution-A was sufficiently mixed with Solution-B, sterile purified water was added to adjust the total volume to 100 mL, and then filtered through a 0.22 μm filter to prepare an ophthalmic composition. The pH of the obtained ophthalmic composition is as described in following Tables 4-5.

TABLE 4

TABLE 5

시험예 1. 물리적 안정성 평가Test Example 1 Physical Stability Evaluation

The ophthalmic compositions prepared in Examples 1 to 25 and Comparative Examples 1 to 5 were stored for 1 week under accelerated storage conditions (40 ± 2 ° C./25% relative humidity or less), and physical stability, ie, appearance and precipitation, was evaluated. , The results are shown in Table 6 below.

TABLE 6

Physical stability assessment

From the results of Table 6, it can be seen that the ophthalmic composition containing vitamin E TPGS as a stabilizer according to the present invention maintained a transparent state without generating precipitate for one week under accelerated storage conditions. In contrast, all of the compositions of Comparative Examples 1 to 5 containing only polysorbate 80 as a stabilizer produced precipitates under accelerated storage conditions, showing low physical stability.

시험예Test Example 2. 화학적 안정성 평가 2. Chemical Stability Assessment

In ophthalmic compositions (ie, ophthalmic compositions prepared in Examples 8, 13, and 23) containing cyclosporin A and sodium hyaluronate, respectively, 0.05 w / v% and 0.30 w / v%, and Comparative Examples 6-10 The prepared ophthalmic composition was stored for 6 months under accelerated storage conditions (40 ± 2 ° C./relative humidity 25% or less), and the chemical stability was evaluated. Chemical stability was measured under the following HPLC analysis conditions for cyclosporin A content at the time of initial, 3 months and 6 months, with each composition stored for 6 months under accelerated storage conditions. The results are shown in Table 7 below.

Column: CAPCELLPAK C18 (4.6 mm X 25 cm)

Column temperature: 80 ℃

Flow rate: 1.2 mL / min

Detector: UV absorbance photometer (wavelength 210 nm)

Injection volume: 10 μl

The peak area of cyclosporin A was confirmed, and the content of cyclosporin A was obtained by the following equation.

Cyclosporine A Content (%)

= Peak area of standard solution / peak area of sample x purity of standard product (%)

TABLE 7

Content of cyclosporin A (%, HPLC peak area)

In the case of eye drop composition, for the approval of the Ministry of Food and Drug Safety, the content of the main ingredient is increased for 6 months under accelerated storage conditions (40 ± 2 ℃ / 25% relative humidity or less), which can replace stability during the expiration date (24 months). 90 to 110% must be maintained. However, from the results of Table 7, it can be seen that the ophthalmic composition of Comparative Examples 6 to 10 containing only polysorbate 80 as a stabilizer does not meet the permit criteria because the content of cyclosporin A is drastically reduced. In contrast, the ophthalmic composition prepared according to the present invention can satisfy the permit requirement of the Ministry of Food and Drug Safety by maintaining the content of cyclosporin A 90% or more for 6 months under accelerated storage conditions.

시험예 3. 안구건조증 유발 마우스 모델을 이용한 약효 평가Test Example 3 Evaluation of Drug Effect Using Dry Eye Induced Mouse Model

Using the ophthalmic composition prepared in Example 8, the therapeutic effect of the cornea damaged by dry eye syndrome was evaluated by corneal staining using a fluorescent dye (Fluorescein) as follows.

(1) Test drug manufacturing

As a test for drug efficacy, a vehicle, a cyclosporin A alone preparation, a sodium hyaluronate alone preparation, a cyclosporine A and a sodium hyaluronate containing preparation (Example 8) were used as shown in Table 8 below. The vehicle, the cyclosporin A-containing formulation and the sodium hyaluronate-only formulation were prepared in the same manner as in Example 8 except for the use of cyclosporin A and sodium hyaluronate, respectively. The content of each component described in Table 8 below represents w / v% unless otherwise indicated.

TABLE 8

(2) dry eye syndrome

Mice were bred from the start of dry eye induction to the end of the experiment (21 days) in a controlled environment chamber (CEC) set at a temperature of 21-23 ° C., a relative humidity of 30% or less, and airflow 15 L / min. 1% atropine sulphate was added twice per day for 2 days (09 o'clock and 21 o'clock), and 2 μl were inoculated into the mouse eye using a micro pipette. Scopolamine was injected subcutaneously into mice at the dose of 0.5 mg / 0.1 mL / head three times / day (09, 13 and 21 hours) from the start of dry eye induction to the end of the experiment.

(3) Evaluation of corneal fluorescein staining

The trigger score was evaluated by corneal fluorescent dye staining on the 3rd day (early) (before the administration of the test substance and before 09 o'clock) after the dry eye was induced for 2 days. Mice were anesthetized with Zoletil 50 (VIRBAC, France) and xylazine (Rompun ^® , Bayer AG, Germany), followed by 2 μl of 1% fluorescein. The conjunctival sac was instilled into the cornea, and after 3 minutes, the cobalt blue light was used to measure and record the staining score of the cornea surface. After determination of the initial trigger score, after separation into four groups (7 per group, n = 7), vehicle, cyclosporin A alone formulation, sodium hyaluronate alone formulation, and Examples in the right eye of each group for 21 days, and Examples Each of the ophthalmic compositions of 8 was administered. On the 7th, 14th and 21st day, the staining score was evaluated in the same manner as the initial induction score evaluation method, and the scores of each group were compared and analyzed. Staining scores were evaluated according to the National Eye Institute (NEI) grading system (ie, FIG. 1).

The scores of the five corneal zones in the National Eye Institute (NEI) grading system of FIG. 1 are as follows.

0: normal (dyed area 0%)

1: Mild (dyed area 0-30%)

2: moderate (dyed area 30 to 60%)

3: Severe (dyed area 60-100%)

Total score: 15

The dry eye syndrome evaluation result performed as described above is shown in FIG. As can be seen from the results of FIG. 2, the ophthalmic composition-administered group prepared according to the present invention showed effective therapeutic activity against corneas damaged by dry eye syndrome, and thus the composition of the present invention can be usefully used as a treatment for dry eye syndrome. have.

Claims

An ophthalmic composition in the form of an aqueous solution comprising cyclosporin and hyaluronic acid or a salt thereof, wherein the ophthalmic composition comprises d-α-tocopherol polyethylene glycol succinate as a stabilizer.
The ophthalmic composition according to claim 1, comprising d-α-tocopherol polyethylene glycol succinate at a concentration of 0.01 to 5.00 w / v%.
The ophthalmic composition according to claim 2, comprising d-α-tocopherol polyethylene glycol succinate at a concentration of 0.10 to 1.00 w / v%.
The ophthalmic composition of claim 1, further comprising at least one stabilizing aid selected from the group consisting of polyoxyl 35 castor oil, polysorbate 80, and polyethylene glycol 200.
The ophthalmic composition according to claim 4, wherein the stabilizing aid comprises polyoxyl 35 castor oil and the polyoxyl 35 castor oil is present at a concentration of 0.10 to 5.00 w / v%.
The ophthalmic composition according to claim 4, wherein the stabilizing aid comprises polysorbate 80 and the polysorbate 80 is present at a concentration of 0.10 to 5.00 w / v%.
The ophthalmic composition according to claim 4, wherein the stabilizing aid comprises polyethylene glycol 200, and the polyethylene glycol 200 is contained at a concentration of 0.10 to 5.00 w / v%.
The ophthalmic composition according to any one of claims 1 to 7, wherein the concentrations of cyclosporin and hyaluronic acid or salts thereof are 0.01 to 2.00 w / v% and 0.01 to 2.00 w / v%, respectively.
The ophthalmic composition of claim 1 comprising 0.05 w / v% cyclosporin, 0.30 w / v% hyaluronic acid or salt thereof, and 0.50 w / v% d-α-tocopherol polyethylene glycol succinate.
5. The method of claim 4, wherein the cyclosporin 0.05 w / v%, hyaluronic acid or its salt 0.30 w / v%, d-α-tocopherol polyethylene glycol succinate 0.50 w / v%, polyoxyl 35 castor oil 1.00 w / v% , And polysorbate 80 0.50 w / v%.
5. The method of claim 4, wherein the cyclosporin 0.05 w / v%, hyaluronic acid or its salt 0.30 w / v%, d-α-tocopherol polyethylene glycol succinate 0.50 w / v%, and polyoxyl 35 castor oil are 2.00 w / v Ophthalmic composition comprising%.
5. Cyclosporin 0.05 w / v%, hyaluronic acid or salt thereof 0.30 w / v%, d-α-tocopherol polyethylene glycol succinate 0.50 w / v%, polyoxyl 35 castor oil 2.00 w / v% , And polyethylene glycol 200 1.00 w / v%.