JP2001514228A - Pharmaceutical composition for treating ocular inflammation containing dexamethasone palmitate - Google Patents

Abstract

  (57) [Summary] Ocular inflammation is treated with an oil-in-water (O / W) emulsion containing an effective amount of a lipophilic ester of dexamethasone.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD OF THE INVENTION

 The present invention provides new formulations and uses thereof for treating ocular inflammation.

[0002]

[Prior art]

In recent years, oil-in-water emulsions have become increasingly important as vehicles for delivering hydrophobic drugs. For example, a fat emulsion described in EP315079 and the like.

[0003] US Pat. No. 4,340,594 describes a parenterally administered fat emulsion containing a steroid with anti-inflammatory activity, in particular dexamethasone palmitate. The fat emulsions described therein are useful in the treatment of rheumatism, immunohemolytic anemia, idiopathic thrombocytopenic purpura, Paget's disease, or treatments associated with kidney transplantation.

In vivo, dexamethasone palmitate acts as a prodrug because it is hydrolyzed by esterases to dexamethasone, a biologically active metabolite thereof. Nevertheless, dexamethasone palmitate has several advantages over conventional water-soluble dexamethasone compositions due to its lipid content.

[0005]

[Problems to be solved by the invention]

The present invention is based on the novel discovery that fat emulsions containing dexamethasone palmitate as an active ingredient are useful in treating ocular inflammation.

Accordingly, the present invention relates to a pharmaceutical composition in the form of an oil-in-water emulsion for the treatment of ocular inflammation, which contains, for example, an effective amount of a lipophilic ester of dexamethasone such as dexamethasone palmitate as an active ingredient. Composition.

[0007]

[Means for Solving the Problems]

The present invention comprises an effective amount of a lipophilic ester of dexamethasone as an active ingredient,
A pharmaceutical composition in the form of an oil-in-water (O / W) emulsion for treating eye inflammation.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION

An emulsion is a dispersion of one liquid in a second immiscible liquid. Since most of the emulsifiers contain water as one of the phases, it is customary to have two types: oil-in-water (O / O) consisting of colloidal particles dispersed in a water column. W) and water-in-oil (W / O) emulsions in which their phases are reversed. A continuous liquid is called a dispersion medium, and a liquid in the form of fine particles is called a dispersed phase.

In order to reach a stable emulsion, a third factor, the emulsifier, must be present. For example, in an O / W emulsion, each colloid particle has an oily core comprising an emulsifier and a surfactant and an outer layer. You containing 10% soybean oil commercially available fatty Emulsions may Intralipid ^TM (Intralipid ^TM; Pharm
acia AB, Sweden).

[0010] Oil-in-water emulsions comprising oil-based particles suspended in an aqueous solvent as described above can incorporate various hydrophobic drugs into their oily cores or their interfacial films, Therefore, it has been proposed and used as a pharmaceutical carrier for such drugs. The present invention relates to the use of an oily ester of dexamethasone as a pharmaceutically active ingredient (i.e., drug).
The present invention relates to the above oil-in-water emulsion for treating eye inflammation.

Typically, the emulsion contains, in addition to the active substance, an oil component and other components, ie, an emulsifier and / or a surfactant. In addition, typically, the aqueous phase (dispersion medium) contains an osmotic agent dissolved therein to raise the osmotic pressure to physiological levels.

Hereinafter, the concentration of the components of the emulsifier is determined by the weight (w / v) of the components in 100 volume units of the total composition.
Is indicated as%.

[0013] The compositions of the present invention comprise an effective amount of a lipophilic ester of dexamethasone. The term "effective amount" is to be understood as an amount sufficient to provide a therapeutic effect. An effective amount in the composition will depend on the dosage form, the regimen (i.e., the composition is given once a day, twice a day, etc.), the age group of the patient, the severity of inflammation, and It obviously depends on various other factors known per se. There will be no difficulty for those skilled in the art to determine by routine and simple experimentation to set an effective amount in each case.

A typical example of a lipophilic ester of dexamethasone is an ester of dexamethasone with a fatty acid, typically such an acid has a chain of 6 to about 22 carbon atoms. Preferred examples of such fatty acids are palmitic, oleic, linoleic,
And stearic acid. A particularly preferred fatty acid is palmitic acid (ie, the activator is preferably dexamethasone palmitate). Typically, dexamethasone palmitate comprises from 0.05%, typically between 0.1% and preferably 0.15%,
It may be included in the composition in a range of 0.5%, typically between 0.4% and preferably between 0.3%.

[0015] The oil composition is typically a vegetable oil. Examples of vegetable oils are oleic acid, linoleic acid, lauric acid, soybean oil, olive oil, safflower oil and the like. Vegetable oils are typically contained at a concentration ranging from about 5% to 30% (w / v).

Typical emulsifiers are phospholipids. Examples of phospholipids are phosphatidylcholine,
Phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine and sphingomyelin. The phospholipid is preferably about 0.5%
It is contained in a concentration range of 10% (w / v), especially in a concentration range of 1-5%.

Typical examples of surfactants are polyalkylene glycols having an average molecular weight of 1,000 to 10,000, polyoxyalkylene copolymers having an average molecular weight of 1,000 to 10,000 and polyoxyethylene fatty acid esters.

Examples of osmotic agents are sucrose or glycerin.

In addition to the above components, the compositions of the present invention may contain various preservatives such as methyl, ethyl and butylparaben, or antioxidants such as α-tocophalol and ascorbic acid.

[0020] The pharmaceutical compositions of the invention will typically be in a suitable dosage form, eg, in the form of eye drops, for example, any drop volume to be administered at each application (eg, 1-2 drops). (Drops).

The present invention also provides a method for treating ocular inflammation comprising topically applying an oil-in-water emulsion containing an lipophilic ester of dexamethasone as an active ingredient to the eye. .

Still further provided by the present invention is the use of a lipophilic ester of dexamethasone and an oil-in-water emulsion for the manufacture of a topical ophthalmic composition for treating ocular inflammation.

Some examples of the present invention are described below, but are not limited thereby.

[0024]

【Example】

1. Experimental method [Animal care] Body weight supplied by a local breeder (Levinstein, Yokneam, Israel) 2.0-
Fifteen male NZW rabbits weighing 2.5 kg were used for the test.

Evaluation of newly developed and clinically applied drugs for treating anterior uveitis, as widely accepted in reports published in scientific publications As the most appropriate experimental model for this study, albino rabbits are used in this study.

Each animal was quarantined on arrival and only appropriate and healthy animals were scheduled for use. Animals were acclimated for 5 days after receiving, during which time they were observed daily for signs of poor health. No abnormalities were detected.

[0027] Rabbits are placed in a laboratory animal facility with restricted access, with a target temperature of 17-23 ° C, 30-7
Maintained at a target humidity of 0% and a 12 hour light / 12 hour dark cycle environmental condition. Temperature and relative humidity were recorded daily. No deviation from the target value was observed.

Rabbits were individually housed in stainless steel cages mounted on batteries. The cage has dimensions of 50 × 45 × 48 cm and is fitted with a stainless steel floor placed over the undertray.

Animals can be uniquely identified by ear number tags.

[0030] Each cage was provided with a front-view cage card containing all details regarding the study number, animal number, gender and animal system, and treatment.

A complete product pelleted rabbit diet (AMBAR 19701) was provided without restriction and the animals had free access to water supplied via a water bottle on the side of the cage.

[0032] 2. Test and Control Compositions Test compositions: Commercially available compositions containing an oil-in-water emulsion containing dexamethasone palmitate at a concentration of 4 mg / ml (LIMETHASON ^™ , Green Cross Corporation)
, Japan).

Reference composition: Commercially available clear aqueous ophthalmic solution containing 1 mg / mol disodium dexamethasone dexamethasone (STERODEX ^™ , Fischcer Pharmaceut)
ical Labs. (1974) Ltd., Israel).

[0034] 3. Experimental Design Conditions [Distribution of Animals to Study Groups] Animals were assigned to study groups in a blind fashion to prevent experimental bias on the part of clinical and laboratory tests. One group was treated with the test composition, one group was treated with the reference composition, and a third group was an untreated control group. The code for each treatment performed on a particular test animal was first revealed at the end of clinical and laboratory experiments.

[Induction of Experimental Uveitis] The EIU administered endotoxin lipopolysaccharide (LPS-E. Coli) at a dose of 2 μg (50 μl LPS / 1 ml HP
CD [hydroxypropylcyclodextrin] was injected intravitreally into the left eye of each test animal (40 μl) for all animals in the three test groups. During treatment, animals were anesthetized by intramuscular injection of 30 mg / kg of ketamine HCL and 5 mg / kg of xylazine HCL.

[EIU Control] The contralateral (right) eye served as a control group for LPS-injected eyes and was similarly injected intravitreally using 50 μl of sterile saline.

[0037] 4. Treatment Immediately following the induction of experimental uveitis, apply one drop of each formulation to both eyes (LPS and saline injected eyes) for both rabbits in the test and reference composition groups The first instillation was repeated 6 times in total. Subsequently, two further instillations were made at intervals of about 3 to 4 hours. The next day, and following the 24 hour clinical study period (see below), the remaining three instillations were performed in a similar manner and at the same intervals.

[0038] 5. Observations and tests Clinical studies Rabbit eye clinical studies are performed 24 and 48 hours after LPS infusion, and according to the grade scale detailed below, conjunctival inflammatory response, iris hyperemia and anterior eye. Based on scoring of tuft flare: I. Grade scale-conjunctiva 0 = normal. The blood vessels of the eyelid and bulbar conjunctiva are easily observed. It may appear bleached to a reddish-brown pink, but without scleral hyperemia. 1 = Red, reddish brown, mainly confined to the eyelid conjunctiva, with some scleral hyperemia mainly confined to the upper and lower parts of the eye. 2 = bright reddish conjunctiva of the eyelid with scleral hyperemia covering at least 75% of the periphery of the scleral area. 3 = beef-like dark red with congestion of both eye and lid conjunctiva, plus significant scleral hyperemia and conjunctival petechiae.

II. Grade scale-iris 0 = normal. An iris without any hyperemia of the iris blood vessels. Occasionally, slightly congested second and third vessels are found near the pupil margin in a small area of about 1-2 mm in diameter. 1 = minimal hyperemia of the second vessel (not in the third vessel). Broad and uniform, but the 1 o'clock or 6 o'clock position may be more pronounced. If the blood vessels confined to that area are clearly hyperemic, the third blood vessel is considered to be substantially hyperemic. 2 = minimal hyperemia of the third blood vessel and minimal to moderate hyperemia of the second blood vessel, or solitary swelling of the iris stroma. 3 = moderate hyperemia of second and third vessels. In addition, some swelling of the iris stroma (most prominent near the 3 and 9 o'clock positions). 4 = significant hyperemia of second and third blood vessels with significant swelling of iris stroma. The iris looks wrinkled and may be accompanied by bleeding in the anterior chamber.

III. Grade scale-anterior chamber flare 0 = normal. There is no flare at all. 1 = slightly detected weak flare. 2 = Medium flare. The iris and lens details are clear. 3 = noticeable iris flare. Iris and lens details are hazy. 4 = Intense flare. Fixed and coagulated aqueous humor with large amounts of fibrin.

The scores from all three clinical parameters measured at each observation period were tabulated separately for each eye of each individual animal. Add the exam scores for 24 and 48 hours and add
It was calculated as "cumulative score."

[Aqueous Aqueous Cell Fraction] Following clinical evaluation of the eyes for 48 hours, the animals were anesthetized (ketamine HCL 100 mg / kg and xylazine HCL 15 mg / kg were injected by im injection) and both eyes of each test animal Was collected using an appropriate hypodermic injection needle and syringe. Cell fractionation was measured using a hemocytometer.

Cell counts were expressed as the total number of fractionated cell types examined (mononuclear and lobulated nuclei; erythrocytes; epithelial cells) in both eyes of individual animals.

[Measurement of protein content of aqueous humor] Immediately after collecting an aqueous humor sample for a cell number test, the remaining aqueous humor was subjected to MIKRO
Centrifuged for 5 minutes in a 12/24 centrifuge (Hettich Zentrifugen). The supernatant was then subjected to a protein assay using a standard commercial assay kit (BioRad Standard III). The protein content of the aqueous humor was measured in both eyes of all test animals.

Preservation of Eyes for Histological Examination Following aspiration of aqueous humor, all test animals were sacrificed and their eyes were excised and fixed with Davidson's solution.

6 Results and Discussion The results of this study are shown below in Figures 1-3 and Table 1; Table 1 summarizes all study group data for clinical treatment, aqueous humor protein content for each of the three treatments. Summarizes the criteria for quantity and cell count.

Table 1: Mean ± S of EIU parameters measured in rabbit left eye (LPS-injected)
List of D

[Table 1]

FIGS. 1-3 show graphs further illustrating the results of Table 1; FIG. 1—Overview of clinical testing criteria; FIG. 2—Aqueous humor protein content; FIG. 3—Aqueous humor cells. number. Each of the bars indicated as A, B, C, D, E and F shows the mean and standard deviation as follows: A, B-untreated control group; A-LPS-injected left eye , And B-right eye with saline injection.

Animals treated with C, D-reference composition; left eye with C-LPS-injection and right eye with D-saline infusion.

Animals treated with E, F-test composition; left eye with E-LPS-injection and right eye with F-saline infusion.

The data for the animals in the untreated control test group show a significant difference in the experimental parameters between the left eye (LPS-injection) and the right eye (saline injection), which is due to the EIU
It reflects the well-known acute inflammatory response to intravitreal LPS infusion in the model. Thus, within 24 hours after intravitreal injection, the left eye of all animals in the group showed significant conjunctival and iris hyperemia and clear clinical signs of anterior chamber flare, which was virtually At the 48-hour consultation. In any of these animals, the right control eye, injected with intravitreal saline, showed no signs of inflammation on two timed examinations.

Measuring both aqueous humor protein content and cell number in the untreated control group confirms clinical observations. In particular, for the former, there is a significant difference between the two contralateral eyes, about 2-4 mg in the control (right) eye.
LPS-injected (left) eyes showed an average protein concentration of approximately 130 mg / ml compared to normal levels of / ml. To assess the degree of acute ocular inflammation, at least in the conditions of the test,
Judgment by total cell number data, this parameter is not shown as a parameter for protein content nor is it a contradictory measure.

[0053] Both the test composition and the reference composition have been demonstrated by effectively reducing the acute inflammatory response caused by EIU and suppressing all three experimental criteria tested. Therefore, regarding the “cumulative score” for evaluating clinical signs, the left (L
(PS-injected) in the untreated control group on the eye in reducing inflammation in the corresponding eye
The relative effect of the two compositions was statistically significant (p <.05). A similar statistically significant (p <.05) treatment effect was also demonstrated by a reduction in aqueous humor protein concentration levels by both compositions.

Regarding the comparative treatment effect of both compositions, a clearly consistently enhanced activity by the test composition was observed in all of the experimental parameters tested. This relative increase was particularly evident from the data for both the "cumulative score" of the clinical evaluation and the aqueous humor protein content. In terms of the percentage difference (%), the test composition treatment led to a reduction in clinical severity and protein content of about 70 and 50%, respectively, while the reference composition treatment was about 55% less. And a corresponding decrease of 35%. This indicates that the optimal concentration of dexamethasone palmitate may be lower than in the test composition, for example 0.2% or 3%.

The treatment applied to the saline-injected control right eye had no apparent effect on either treatment.

[Brief description of the drawings]

FIG. 1 is a bar graph (mean ± S) showing the overall score of the clinical trial.
D).

FIG. 2 is a bar grab (mean ± SD) showing aqueous humor protein content.

FIG. 3 shows bar graphs showing aqueous humor cell counts for various treatments; control (A, B), treatment with reference composition (C, D) and treatment with the composition of the present invention.
(E, F). In each case, the left eye was injected with LPS (results are shown as columns A, C, and E in each of FIGS. 1-3), and the right eye was injected with saline (results were: In each of FIGS. 1-3, shown as columns B, D and F).

[Procedural Amendment] Submission of translation of Article 34 Amendment of the Patent Cooperation Treaty

[Submission date] February 28, 2000 (2000.2.28)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Claims

[Correction method] Change

[Correction contents]

[Claims]

──────────────────────────────────────────────────続 き Continuation of front page (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE ), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE, GH, GM, HR, HU, ID, IL, IS, JP, KE, KG, KP , KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, US, UZ, VN, YU, ZW

Claims (8)

[Claims] 1. A pharmaceutical composition in the form of an oil-in-water (O / W) emulsion for treating ocular inflammation, comprising an effective amount of a lipophilic ester of dexamethasone as active ingredient. 2. The pharmaceutical composition according to claim 1, wherein the active ingredient is an ester of dexamethasone and a fatty acid. 3. The pharmaceutical composition according to claim 2, wherein the ester of dexamethasone is dexamethasone palmitate. 4. The pharmaceutical composition according to claim 1, which is in the form of eye drops. 5. Use of a lipophilic ester of dexamethasone and an oil-in-water (O / W) emulsion for the manufacture of a topical ophthalmic composition for treating ocular inflammation. 6. The use according to claim 5, wherein the dexamethasone ester is dexamethasone palmitate. 7. A method for treating eye inflammation comprising topically applying an oil-in-water (O / W) emulsion containing an lipophilic ester of dexamethasone as an active ingredient to the eye. 8. The method of claim 7, wherein the ester of dexamethasone is dexamethasone palmitate.