United States Patent [191
`Ogawa et al.
`
`[11] Patent Number:
`[45] Date of Patent:
`
`‘r 4,910,225
`Mar. 20, 1990
`
`[54] LOCALLY ADMINISTRABLE
`THERAPEUTIC COMPOSITION FOR
`INFLAMMATORY DISEASE
`
`[75] Inventors: Takahiro Ogawa, Nishinomiya;
`Yoshikazu Kurihayashi, Kobe;
`Kazumichi Ushio, Nishinomiya;
`Akira Ohtori, Nara, all of Japan
`
`[73] Assignees: Seniu Pharmaceutical Co., Ltd.,
`Osaka, Japan; A. H. Robins
`Company, Incorporated, Richmond,
`Va.
`
`OTHER PUBLICATIONS
`Walsh et al., Journal of Medicinal Chemistry, 1989, vol.
`27, No. 11, pp. 1379-1388.
`Chem. Abst. l07—(1987)-211870z.
`Primary Examiner-Stanley J. Friedman
`Attorney, Agent, or Firm-Wenderoth, Lind & Ponack
`[57]
`ABSTRACT
`This invention relates to a locally administrable thera
`peutic composition for inflammatory disease which is
`characterized by comprising benzoylphenylacetic acid
`of the formula
`'
`
`[21] Appl. No.: 301,033
`
`[22] Filed:
`
`Jan. 24, 1989
`
`Foreign Application Priority Data
`[30]
`Jan. 27, 1988 [JP]
`Japan ................................ .. 63-16683
`
`[51] Int. C13 .......................................... .. A61K 31/195
`
`[52] US. Cl. . . . . . . . . . . . . . . . .
`
`. . . . . . . . . . .. 514/561
`
`[58] Field of Search ....................................... .. 514/561
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`4,045,576 8/ 1977 Welstead et al. ................. .. 424/309
`4,126,635 11/1978 Welstead et al.
`562/441
`4,568,695 2/1986 Moran et al. ...... ..
`514/648
`4,683,242 7/1987 Poser ................................. .. 514/539
`
`FOREIGN PATENT DOCUMENTS
`
`0221753 5/1987 European Pat. Off. .
`58-201710 2/1984 Japan.
`
`I
`
`NH;
`
`CHzCOOl-l
`
`(wherein R is a hydrogen or halogen atom), or a salt
`thereof, or the hydrate of said acid or salt, as active
`ingredient.
`An ophthalmic composition according to the invention
`can treat effectively in?ammatory eye disease by topi:
`cal application, is not an irritant to the eye, and has a
`superior effect to conventional drugs of the same or
`similar type.
`The aqueous composition prepared in accordance with
`this invention has excellent stability and can be used
`advantageously as a nasal or otic composition as well as
`an ophthalmic one in the treatment of in?ammatory otic
`or nasal disease.
`
`9 Claims, No Drawings
`
`LUPIN EX1004, Page 1
`
`

`
`1
`
`LOCALLY ADMINISTRABLE THERAPEUTIC
`COMPOSITION FOR INFLAMMATORY DISEASE
`
`4,910,225
`2
`topical application, and that the effectiveness of such
`drugs is compatible with that of conventional steroid
`anti-in?ammatory drugs.
`Furthermore, since the inventors obtained the ?nding
`that there are some problems that the above-mentioned
`benzoylphenylacetic acid derivatives are unstable in an
`aqueous solution with the optimal pH range for a lo
`cally administrable therapeutic composition, they ex
`tensively investigated in search of the method for the
`preparation of a stable aqueous solution. As a result, we
`have succeeded in preparing a stable aqueous composi
`tion. Thus, the stable aqueous composition according to
`the invention are achieved based on the above ?nding.
`While a number of non-steroid compounds fall under
`the category of anti-inflammatory agents, all of them
`are not effective in treating in?ammatory eye diseases
`when topically administered to the eye. This is because
`there are several problems lying before them. First,
`when topically administered to the eye, a medicinal
`agent has to pass through the cornea so that it can reach
`the site of in?ammation. Even when it has succeeded in
`arriving at the site of in?ammation, it must remain there
`in a necessary concentration for a necessary period of
`time. If it fails to meet these requirements, it will be
`unable to produce expected therapeutic effects. Fur
`thermore, in case it is irritative to the eye, it is rather
`possible that the topical administration of the medicinal
`agent to the eye would cause exacerbation of symptoms.
`Therefore, great caution and much care are necessary in
`selecting a medicinal agent for topical administration to
`the eye. Furthermore, in case of administration in the
`form of eye drops, it goes without saying that it is desir
`able that the eye drop is stable for a long period of time
`in an aqueous solution without decomposition or form
`ing insoluble matters.
`.
`Accordingly, it is an object of the invention to solve
`the above problems and provide a novel and useful
`agent for ophthalmic use.
`I
`-
`Moreover, the other object of the invention is to
`provide a suf?ciently stable aqueous solution
`such as
`eye drops, otic
`solution and nasal solution which con
`tains the above compounds when stored for a long per
`iod of time.
`
`25
`
`BACKGROUND OF THE INVENTION
`1. Field of the Invention
`This invention relates to a locally administrable ther
`apeutic composition for in?ammatory eye disease as
`well as nasal or otic in?ammatory disease.
`More particularly, it relates to a locally administrable
`therapeutic composition for in?ammatory eye as well as
`for nasal or otic in?ammatory disease, which contains
`as active ingredient a benzoylphenylacetic acid deriva
`tive, a salt thereof or the hydrate of said acid or salt.
`The other object of the present invention is to pro
`vide a stable locally administrable aqueous composition
`such as eye drop, otic composition and nasal composi
`tion containing the above compounds.
`2. Description of the Prior Art
`That certain benzoylphenylacetic acid derivatives,
`when orally administered, exhibit anti-in?ammatory
`activity has been reported in detail in Journal of Medici
`nal Chemistry, Volume 27, pages 1379-1388 (1984),
`among others. Furthermore, Japanese Laid Open Pa
`tent Publication No. l26l24/ 1987 describes pharmaceu
`tical compositions for percutaneous administration
`which contain these compounds. However, none of the
`published literature-inclusive of the above-mentioned
`patent speci?cation-contains any description indicating
`or suggesting that these medicinal substances are effec
`tive against in?ammatory disease of the eye, nose or car
`when they are administered topically.
`For the treatment, with topical application of drugs,
`of in?ammatory ophthalmopathy such as uveitis and
`conjunctivitis which are most frequently observed in
`the ophthalmological ?eld, steroid drugs such as dex
`amethazone have so far been employed. Topical appli
`cation of steroid drugs to the eye has some apprehen
`sion of increasing intraocular pressure to cause glau
`coma. And, there is a fear not only of causing corneal
`perforation when such steroid drugs are applied to pa
`tients suffered from corneal herpes, corneal ulcer or the
`like, but also of induction of corneal herpes, keratomy
`cosis, Pseudomonas infections and the like by the topi
`cal application of steroid drugs. As there has been
`known such-effects as above, steroid anti-in?ammatory
`agents shall be applied with particular care. In spite of
`such a situation, there has not been known any non-,
`steroid anti-in?ammatory agent compatible with steroid
`anti-in?ammatory drugs in effectiveness for the treat
`ment of in?ammatory opthalmopathy such as uveitis.
`Thus, in the present stage in this technical field, for the
`treatment of in?ammatory ophthalmopathy, it is hardly
`possible not to use steroid anti-in?ammatory agents
`with particular care to avoid the side effects as above
`mentioned. Under such circumstances, it is natural that
`ophthalmological experts are
`awaiting the appearance
`of non-steroid drugs which are effectively usable
`against uveitis or the like.
`The present inventors investigated to find out topi
`
`cally applicable drugs with lesser side-effects
`and with
`superior effectiveness by which topically applicable
`drugs having been employed in the treatment ofin?am
`matory ophthalmopathy, i.e. steroid anti-in?ammatory
`agent, can be replaced. As a result, the present inventors
`unexpectedly found that certain derivatives of benzoyl
`phenylacetic acid are very effective in the treatment of
`in?ammatory ophthalmopathy, especially of uveitis, by
`
`45
`
`55
`
`65
`
`SUMMARY OF THE INVENTION
`The present
`invention, which has been completed
`based on the above finding, provides a therapeutic com
`position for administration to the eye for the treatment
`of in?ammatory eye diseases which contains as active
`ingredient a benzoylphenylacetic acid of the formula
`
`0
`II
`c
`
`[wherein R is a hydrogen or halogen atom], or salt
`thereof, or the hydrate of said acid or salt. In the for~
`mula. the halogen atom represented by R is, for exam
`ple, ?uorine, chlorine. bromine or iodine. The above
`compound to be used in accordance with the invention
`may be in a salt form. The salt includes alkali metal salts
`such as sodium salt and potassium salt, alkaline earth
`metal salts such as calcium salt and magnesium salt,
`among others, and any salt may suitably be used pro
`
`LUPIN EX1004, Page 2
`
`

`
`3
`vided that it can attain the object of the invention. The
`compounds de?ned above may be obtained in the form
`of a hydrate depending on the conditions of synthesis,
`recrystallization and so forth, and such form may be
`used in practicing the invention without any inconve
`nience or trouble.
`Further, the above compounds may be unstable when
`stored in an aqueous solution for a long period of time,
`and there are some problems in the stability of an aque
`ous solution containing the compounds. Therefore the
`inventors extensively investigated the stabilizing
`method in order to enhance the stability. As a result,
`unexpectedly, they have succeeded in stabilizing the
`solution by incorporating a water-soluble polymer and
`sul?te and adjusting the pH to about 6-9.
`
`10
`
`20
`
`25
`
`40
`
`45
`
`30
`
`4,910,225
`4
`tion may further contain pharmaceutically active ingre
`dients, such as an anti-in?ammatory agent of another
`kind, an analgesic and an antimicrobial, unless they are
`un?t for the purpose of attaining the object of the inven
`tion. Examples of such antiin?ammatory agent are indo
`methacin and pranoprofen. Usable examples of the anti
`microbial agents are penicillins, cephalosporins, and
`synthetic antimicrobial agents of the quinolonecarboxy
`lic acid series. Among these active ingredients for com
`bined use with the active ingredient according to the
`invention, the anti-in?ammatory agent is expected to be
`synergistic with said active ingredient in the ophthalmic
`compositions according to the invention. The analgestic
`is suited for the purpose of alleviating inflammation
`associated pain, and the antimicrobial agent is suited for
`the purpose of preventing secondary infection. It is of
`course possible to incorporate active agents other than
`those mentioned above in the ophthalmic compositions
`according to the invention unless the object of the in
`vention cannot be attained due to the presence thereof.
`In preparing the ophthalmic compositions according
`to the invention as mentioned above, an isotonizing
`agent, a microbicidal agent or preservative, a chelating
`agent, a thickening agent and so forth may be added to
`the compositions in accordance with the general prac
`tice of ophthalmic preparation manufacture. The isoto
`nizing agent includes, among others, sorbitol, glycerine,
`polyethylene glycol, propylene glycol, glucose and
`sodium chloride. The preservative includes para
`oxybenzoic acid esters, benzyl alcohol, parachloro
`meta-xylenol, chlorocresol, phenetyl alcohol, sorbic
`acid and salts thereof, thimerosal, chlorobutanol, and
`the like. The chelating agent is, for example, sodium
`edetate, sodium citrate or sodium salt of condensed
`phosphoric acid. In preparing the ophthalmic composi
`tions according to the invention in the form of eye
`ointments, the ointment base can be selected from
`among petrolatum, Macrogol, carboxymethylcellulose
`sodium, etc.
`'
`The ophthalmic composition according to this inven
`tion is prepared by incorporating the active compound
`in a base or vehicle for topical application to the eye. To
`prepare a liquid preparation, the concentration of the
`active ingredient may range from about 0.001% to
`about 10% and is preferably in the range of about 0.01%
`to about 5%. An ointment may be prepared by using the
`active compound in a concentration from about 0.001%
`to about 10%, preferably about 0.01% to about 5%. The
`ophthalmic composition of this invention may be ad
`ministered in accordance with the following schedules.
`In the form of eye-drops, one to several drops per dose
`are instilled with a frequency of once to 4 times a day
`according to the clinical condition. Of course, the dos
`age may be adjusted according to symptoms. The oph
`thalmic composition according to this invention can be
`used topically for the treatment of inflammatory dis
`eases of the eye without causing local irritant effects
`and produces bene?cial effects surpassing those obtain
`able with the conventional drugs of the same type.
`According to this invention. there can be obtained a
`stable aqueous composition such as otic composition or
`nasal composition. Other conventional methods can be
`used unless unsuitable for the object of this invention.
`Among others, an isotonizing agent. buffer solution and
`preservatives can be used. The concentrations of the
`compounds of the invention varies depending on symp
`toms and so on, and usually may be in the range of about
`0.001 to_ about 10%, preferably about 0.01 to about 5%.
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`The compounds to be used as active ingredients in the
`topically administrable therapeutic compositions for
`in?ammatory eye disease as well as nasal or otic disease
`in accordance with the invention (although such com
`positions are occasionally hereinafter referred to as
`“opthalmic composition according to the present inven
`tion", use of this abbreviation does not exclude the
`application of the composition in the nasal or otic ?elds)
`can be produced as described in the above-cited report
`in Journal of Medicinal Chemistry, Volume 27, pages
`1379-1388 (1984) or U.S. Pat. No. 4,045,576, for in
`stance, or by a modi?cation of the method described
`therein. The ophthalmic compositions according to the
`invention can be prepared in the form of eye drops, eye
`ointments and so on in the same manner as various
`known compositions for topical administration to the
`eye. Thus, a compound of the above formula or a mix
`ture of two or more compounds of the above formula is
`preferably made up into an aqueous or non-aqueous
`solution or mixed with an ointment base suited for oph
`thalmic use. On that occasion, an aqueous base gener
`ally used in the production of ophthalmic preparations,
`for example sterile distilled water, is suitably used as the
`aqueous base and the pH thereof is adjusted to a level
`suited for topical administration to the eye. It is desir
`able that an appropriate buffer should be added in ad
`justing the pH. The pH of the ophthalmic compositions
`according to the invention is selected with due consid
`eration paid to the stability and topical eye irritativity of
`the active ingredient, among others. According to the
`present invention, the stability of an aqueous composi
`tion containing the above compounds is remarkably
`enhanced by incorporating a water-soluble polymer and
`sul?te, and adjusting the pH to 6.0-9.0, preferably about
`7.5-8.5. The eye irritation of the solution is not ob
`served. A water-soluble polymer includes polyvinyl
`pyrrolidone, carboxypropylcellulose, hydroxyethylcel
`lulose, hydroxypropylcellulose, polyvinyl alcohol, so
`dium salt of polyacrylic acid an so on. Polyvinyl pyrrol
`idone is preferred among them. The concentration of a
`water-soluble polymer is in the range of about 0.1 to 10
`w/w%. Sul?te includes sodium, potassium, magnesium,
`calcium salt and so on. The concentration of sulfite is in
`the range of about 0.1 to L0. w/w %. The pH adjust
`ment is generally conducted with sodium hydroxide or
`hydrochloric acid, for instance, and it is advisable to
`form a buffer solution by combined use of, for example.
`sodium acetate, sodium borate or sodium phosphate and
`acetic acid, boric acid or phosphoric acid, respectively.
`The ophthalmic compositions according to the inven
`
`55
`
`60
`
`65
`
`LUPIN EX1004, Page 3
`
`

`
`4,910,225
`6
`5
`and the concentration of prostaglandins in the anterior
`The following experimental examples are given to
`delineate the ef?cacy pro?le of the ophthalmic compo
`chamber aqueous humor.
`Administration of the test drug
`sition of this invention and the stability of the aqueous
`Gross observation was made on the day after injec
`compositions of the invention.
`tion of BSA into the vitreous body and with the animals
`arranged in the decreasing order of severity of ocular
`inflammation, grouping was carried out in such a man
`ner that the intensity distribution would be uniform
`over the groups. Thus, a physiological saline group of 7
`animals, a 0.1% Compound [I] instillation group of 4
`animals and a 0.5% Compound [I] instillation group of
`5 animals were provided. After this grouping proce
`dure, the test drugs and saline were respectively in
`stilled into both eyes of the rabbits, 50 ul per dose, 4
`times a day. For induction of ophthalmitis II, each drug
`was instilled into both eyes, 50 [1.1 per dose, immediately
`after injection of BSA into the auricular vein and at
`l-hour intervals thereafter, for a total of 14 times.
`
`EXPERIMENTAL EXAMPLE 1
`Anti-in?ammatory effect of the ophthalmic agent
`according to this invention in experimental ophthalmitis
`induced by bovine serum albumin in white rabbits
`[Animals]
`Seventeen male white rabbits weighing about 2 kg
`were used. They were fed with 80 g of Labo RG-RO
`(Nippon Agricultural Co., Ltd.) daily and had free ac
`cess to tap water.
`
`10
`
`[Test drug]
`Sodium 3-(4-bromobenzoyl)~2-aminophenylacetate
`monohydrate (hereafter referred to as Compound [I]
`was used as 0.5% and 0.1% ophthalmic solutions. These
`ophthalmic solutions had a pH value of 8.11 and osmo
`larities of 310 mOsm/kg-HZO and 325 mOsm/kg-I-IgO,
`respectively. Bovine serum albumin (hereafter referred
`to as “BSA“) was dissolved in physiological saline to a
`concentration of 5% and sterilized by ?ltration. A 0.1
`ml portion of the solution was injected into the central
`part of the vitreus of both eyes using a 27G needle
`under anesthesia with 0.4% oxybuprocaine hydrochlo
`ride to induce ophthalmitis (ophthalmitis I). After 28
`days when ophthalmitis I had nearly recovered, 2.5%
`BSA solution was administered in a dose of 25
`mg/ml/kg into the auricular 'vein to cause ophthalmitis
`(ophthalmitis II). The severity of ophthalmitis was
`rated according to the rating scale") of Yamauchi et al.,
`based on the Draize method in which an increased
`weight given to the internal segment of the eye. Obser
`vation was made with a frequency of once in one or two
`days during the peak period of in?ammation and once
`in three or four days before and after the peak period for
`ophthalmitis I and 3, 6, l2 and 24 hours after intrave
`nous injectton of BSA for ophthalmitis II.
`m Hideyasu Yamnuchi. Mukoto lngu. Tadashi lso and K020 Uda:
`Anti-in?ammatory effect of ?uorometholone ophthalmic solution in
`experimental uveitis in rabbits. Folia Ophthalmologica Japonica. 24:
`969-7‘) (I973).
`
`20
`
`30
`
`35
`
`45
`
`Evaluation of results
`' In ophthalmitis I, Compound [I] at concentrations of
`0.1% and 0.5% caused a potent and dose-dependent
`inhibition for both the external and the internal segment.
`of the eye. Furthermore, at both concentration levels,
`Compound [I] produced a substantially complete inhibi
`tion of prostaglandins in the aqueous humor in ophthal
`mitis I.
`In regard to the inhibitory effect on in?ammatory
`symptoms, as evaluated by gross observation, which are
`induced by the intraveous injection of antigen, the
`Compound [I] according to this invention produced a
`substantially complete inhibition at both concentrations.
`As to white blood cell count, all drugs produced nearly
`the same degree of inhibition in both the internal and
`the external segments of the eye.
`For any of the drugs, no body weight suppression
`was observed even after 28 consecutive days of treat
`ment. In the organs including the thymus, spleen, adre
`nal and so on, anatomically no abnormality was found.
`
`EXPERIMENTAL EXAMPLE 2
`The effect of the compounds according to this
`invention on carrageenin edema in rats
`Test drugs
`1. Sodium 3-(4-bromobenzoyl)-2-aminophenyl-acetate
`(hereinafter referred to as Compound [1])
`2. Sodium 3-(4-chlorobenzoyl)-2-aminophenyl-acetate
`(hereinafter referred to as Compound [11])
`3. Sodium 3-benzoyl-Z-aminophenylacetate (hereinafter
`referred to as Compound [1111)
`
`[Results]
`Anti-in?ammatory effect in ophthalmitis I
`Table I shows the sum of scores for respective pa
`rameters during a peak in?ammatory period of 3 days
`after aseptic injection of 5% BSA into the central part
`of the vitreous.
`Table 2 shows the amount of protein, white blood cell
`count and the concentration of prostaglandins in the
`anterior chamber aqueous humor.
`Anti-in?ammatory effect in ophthalmitis II
`The administration of 2.5 ml/kg of 2.5% BSA solu
`tion into the auricular vein after 29 days when the in
`?ammatory symptoms
`of ophthalmitis I had substan
`tially subsided resulted in a relapse of in?ammation after
`3 hours in the physiological saline group, where both
`the external and internal segment of the eye after 12
`hours showed in?ammatory pictures similar to those
`observed at the peak of ophthalmitis I. These symptoms
`were still observed even after 24 hours. Table 3 shows
`the scores for
`respective parameters at 3, 6, l2 and 24
`hours after the intraveous injection of BSA. Table 4
`shows the amount of protein, white blood cell count
`
`55
`
`65
`
`Method
`Using female Wister rates weighing l00 g in groups
`of 5 animals or 10 eyes, 0.05 ml of 1% carrageenin
`(dissolved in physiological saline at 50° C.) as a phlogo
`gen was injected beneath the conjuctiva of both eyes to
`induce edema. Physiological saline, as a control, and
`test drugs were respectively instilled into both eyes 40
`and 20 minutes before and immediately after the injec
`tion of carrageenin, in the amount of 2.5 ul per dose.
`Four hours after the phlogogen treatment, each animal
`was sacrificed by cervical dislocation and in accordance
`with the method of Maistrello et al.<3l, the scalp was
`peeled off toward the eyelid and the edematous portion
`together with the skin was removed along the lid mar
`gin and weighed. The degrees of inhibition of carra
`geenin edema in the control group and drug treatment
`
`LUPIN EX1004, Page 4
`
`

`
`7
`group are shown in Table 5. Each drug group showed
`a signi?cant difference from the control group, indicat
`ing the effectiveness of the three compounds against
`acute ocular in?ammation.
`(Z) Maistrello et aL: Quantitative Effect of Topically Applied Anti-in‘
`?ammatory Agents on External Ocular In?ammation in Rats: Journal
`of Pharmaceutical Science, volume 62, pp. 1455-6009711).
`
`4,910,225
`8
`Compound [I] has stronger anti-in?ammatory effect
`than indomethacin.
`
`EXPERIMENTAL EXAMPLE 4
`
`5
`
`Formula
`Compound [I]
`Borax
`Sodium borate
`Sodium chloride
`Disodium edetate
`Benzalkonium chloride
`Polysorbate 80
`Sterile puri?ed water
`
`0.1 g
`1.0 g
`Sufficient quantity
`0.25 g
`0.02 g
`0.005 g
`0.3 g
`To make 100 ml
`
`Stability was observed at 60° C. of the compound by
`changing pH (6.0, 7.0, 8.0 and 9.0) of the above formula.
`The results are shown in Table 8.
`Of the above four, the formula at the pH of 8 is most
`stable. In the formula, the change in residue rate were
`not almo'st observed but in three weeks red insoluble
`matters were observed.
`EXPERIMENTAL EXAMPLE 5‘
`As a result of extensive examination on preventing
`the red insoluble matters, the stability was observed by
`incorporating polyvinyl pyrrolidone.
`
`Formulas
`
`Compound [I]
`Boric acid
`Borax
`Disodium edetate
`Benzalkonium chloride
`Polysorbate 80
`Polyvinyl pyrrolidone
`Sterile purified water
`
`B-1 7
`
`8-2
`
`0.1 g
`0.1 g
`1.5 g
`‘1.5 g
`Sufficient quantity
`0.02 g
`0.02 g
`0.007 g
`0.007 g
`0.15 g
`0.15 g
`2.0 g
`—
`To make 100 m1
`.pH 8
`pH 8
`
`In the above formulas, the results of the stability
`60° C. are as follows (Table 9):
`It was found that by incorporating polyvinyl pyrrol
`idone, the appearance of red insoluble matters was con
`siderably prevented. In four weeks, however, some
`insoluble matters were observed.
`
`at
`
`EXPERIMENTAL EXAMPLE 6
`Moreover, as a result of searching for more stable
`solutions, the inventors obtained the ?nding that by
`further incoporating sodium sulfite other than polyvinyl
`pyrrolidone, the stability was remarkably increased.
`
`Formulas
`
`Compound [I]
`Boric acid
`Borax
`Disodium edetute
`Benzalkonium chloride
`Polyvinyl pyrrolidone
`Sodium sul?te
`Sterile puri?ed water
`
`B
`
`8-3
`
`0.1 g
`0.1 g
`1.5 g
`1.5 g
`Sufficient quantity
`0.02 g
`0.02 g
`0.007 g
`0007 g
`0.15 g
`0.15 g
`—
`0.2 g
`To make 100 ml
`pH 8
`pH 8
`
`As shown in Table 10, the change of appearance was
`observed in the formula in which sodium sul?te was not
`incorporated. and the residue increased by about 7%.
`By contrast, In the solution containing Compound [I] in
`which polyvinyl pyrrolidone and sodium sul?te coexist,
`the change of appearance was not observed at all and
`
`EXPERIMENTAL EXAMPLE 3
`Effects on atropine-resistant miosis and on protein
`increase after paracentesis
`The experiment was divided into two parts, i.e. Ex
`periment 3.0, in which the effect of Compound [I] was
`evaluated, and Experiment 3.b, in which indomethacin,
`the most known anti-in?ammatory drug with strong
`cyclooxgenase inhibitory activity, was evaluated.
`[Test drugs]
`The solutions of the following formulas were used.
`
`15
`
`MEL
`Compound [I]
`Boric acid
`Borax
`Sodium chloride
`Sodium edetate
`Benzalkonium chloride
`Tween 80
`
`0.0001%
`0.001
`0.01
`0.1
`1.0%
`1.0
`1.0
`1.0
`q.s.
`q.s.
`q.s.
`q.s.
`0.25%
`0.25
`0.25
`0.25
`0.02%
`0.02
`0.02
`0.02
`0.005%
`0.005
`0.005
`0.005
`0.3%
`0.3
`0.3
`0.3
`(pH 8.0, Osmotic pressure 310 mOsm/KgHZO)
`b. Indomethacin
`Indomethacin
`Castor oil
`
`0.5%
`q.s.
`
`20
`
`25
`
`30
`
`35
`
`45
`
`50
`
`55
`
`60
`
`65
`
`[Animals]
`Totally 28 male albino rabbits (4 rabbitsX7 groups)
`with a body weight of about 2 kg werr used. They had
`been con?rmed, before the experiment, to have mydri
`atic response to 1% atropine for at least 4 hours.
`[Test procedure]
`50 ul each of 1% atropine solution was instilled into
`both eyes of the animals one hour before the 1st para
`centhesis, in which approx. 0.2 ml/eye of aqueous
`humor (primary aqueous humor) was collected. Topical
`application of 50 pl each of the test drug solutions was
`conducted 30 min before the paracenthesis. Pupil diam
`eter of each eye was measured with a slide caliper im
`mediately before and 10 min after the paracenthesis.
`The 2nd paracenthesis was conducted 90 min after the
`1st one, in which approx. 0.2 ml/eye of aqueous humor
`(secondary aqueous humor) was collected.
`[Results]
`As shown in Table 6, Compound [I] exhibited a dose
`related inhibitory activity on miosis after paracentesis at
`the concentrations of 0.0001—0.1%, whereas little effect
`was observed with indomethacin at the concentrations
`as high as 0.5%. As shown in Table 7, Compound [I]
`exhibited a strong and dose-related inhibitory activity
`on protein increase after paracenthesis, in which the
`effect of 0.01% of Compound [I] was equivalent to that
`of 0.5% indomethacin. It is well known that atropine
`resistant miosis and protein increase in aqueous humor
`after paracentesis are caused by prostaglandin E1,
`which—is one of the most important chemical mediators
`of in?ammation and is synthesized immediately after
`mechanical injury. The results, therefore, indicate that
`
`LUPIN EX1004, Page 5
`
`

`
`9
`the decomposition of Compound [I] was not observed
`either. It was found that the stability was remarkably
`enhanced. Thus, there can be successfully obtained a
`stable aqueous composition containing the compounds.
`The following are explanatory examples of the oph- 5
`thalmic composition and other stable aqueous composi
`tions according to the invention.
`
`4,910,225
`10
`The above ingredients are made up into an ophthal
`mic solution (the total volume being 100 ml) and the pH
`is adjusted to 7.5 with hydrochloric acid.
`
`EXAMPLE 6
`Ophthalmic Solution
`
`EXAMPLE 1
`
`.
`
`.
`
`:coedéluti: r5:ISL-)1:rodipaotléenzoyl)-2-aminophenyl-
`B .
`.d
`y
`Bgilatit am
`.
`.
`Sodium chloride
`_
`sodlum “lame
`Benzalkonium chloride
`Polysorbate 80
`Puri?ed water
`
`Sodium 3-(4-bromobenzoyl)-2-aminophenyl-
`10 acetate monohydrate
`Boric acid
`Borax
`
`Disodium edema
`Benzalkonium chloride
`Polysorbate 80
`Polyvinyl pyrrolidone
`15 Sodium sun-"e
`Sterile puri?ed water
`PH 8
`
`0.1 g
`
`l.25 g
`1.0 g
`
`002 g
`0.005 g
`0'15 g
`2.0 g
`01 g
`To make 100 ml
`
`0.l%
`l W
`Suf?cienf quaantity
`0.25%
`002%
`0.005%
`0.3%
`Sufficient quantity
`
`h h l 20
`The above in redients are made u into an o t a -
`-
`.
`g
`-
`p
`p
`-
`mic solution (the total volume being 100 ml) and pH is
`adjusted to 8.0.
`
`EXAMPLE 7
`.
`.
`Ophthalmic Solution
`
`EXAMPLE 2
`
`Sodium S-benzoyl-Lamino-phenyl-
`acetate
`Boric acid
`Borax
`Sodium chloride
`Sodium edetate
`Benzalkonium chloride
`po|ysorbme 30
`Puri?ed water
`
`0.l%
`
`l.0%
`0.02%
`0.25%
`Suf?cient quantity
`0.005%
`(13%
`Suf?cient quantity
`
`The above ingredients are made up into an ophthal-
`mic solution (the total volume being 100 ml) and pH is
`adjusted to 8.0.
`
`EXAMPLE 3
`
`_ _~
`
`.
`.
`Sodium 3 (4 chlorobenzoyl) .. amino
`phenylacetaie
`wh.!
`I.
`l C P?ll'o alum
`
`.7.-
`
`.
`
`l.0%
`
`,
`
`I.‘
`_
`S fr‘. t
`U lLlCn quan l y
`
`The above ingredients are mixed up into an eye oint-
`ment (100 g) in the conventional manner.
`
`EXAMPLE 4
`
`s d‘
`szdlzmnmbenzoyl)_Z_umin0_
`phenymcewe monohydme
`Carboxymelhylccllulose
`
`001
`'
`
`g
`
`Sufficient quantity
`
`The above ingredients are mixed up in the conven-
`tional manner to give 100 g of an eye ointment.
`
`Sodium 3-(4-bromobenzoyl)-2-aminophenyl-
`25 acetate monohydrate
`~
`Boric acid
`Borax
`Sodium chloride
`Polysorbate 80
`Methylparaben
`30 Ethylparaben
`Polyvinyl pyrrolidone
`sodium Sul?le
`Sodium edelale
`Sterile puri?ed water
`
`0.1 g
`
`0.7 g
`Sufficient quantity
`0.5 g
`0.15 g
`0.013 g
`0.007 g
`2.0 g
`0.2 g
`002 g
`To make 100 ml
`
`35
`
`EXAMPLE 8
`Ophthalmic Solution
`
`40 Sodium 3-(4-bromobenzoyl)-2-aminophenyl-
`acetate monohydrate
`Boric acid
`Borax
`Bcnzulkonium chloride
`
`Polysorbate 80
`Polyvinyl pyrrolidone
`Sodium sul?te
`2:116 pun?ed water
`
`0.1 g
`
`l.5 g
`Suf?cient quantity
`0005 g
`
`015g
`g
`0.1 g
`To make 100 ml
`
`50
`
`The following (Table ll) are the residue and appear
`ance of the com ositions in Exam les 6-8 after 4 weeks
`P
`P
`at 60° c’
`As shown in Table 11, it was found that changes in
`appearances of the compositions were not observed at
`55 all, and the decomposition Of the compound was not
`almost observed, the aqueous compositions being stable,
`excellent for a long period of time‘
`EXAMPLE 9
`
`EXAMPLE 5
`
`6O
`
`Ophthalmic Solution
`
`Sodium
`3-(4-chlorohenzoyl)-Z-amino-
`phcnylacetate monohydrute
`Sodium chlmidc
`Tween 80
`Puri?ed water
`
`1_() g
`
`()_8 g
`()2 g
`Suf?cicnt quantity
`
`Sodium 3-(4-hromobenzoyl)-Z-aniinophenyl-
`acetate monohydl'ale
`Sodium monohydrogen phosphate
`65 Sodium dihydrogen phosphate
`Sodium chloride
`ncnzulk‘mium chlmidc
`l’olysorbatc 80
`Polyvinyl alcohol
`
`0.1 g
`
`0.2 g
`Sul'?cient quantity
`0.8 g
`0-007 g
`0.15 g
`1.0 g
`
`LUPIN EX1004, Page 6
`
`

`
`11
`-continued
`
`4,910,225
`
`12
`-continued
`
`Potassium sul?te
`Sterile puri?ed water
`pH 8
`
`0.2 g
`To make 100 ml
`
`EXAMPLE 10
`-
`-
`Nasal and one Solunon
`
`Sodium 3-(4-bromobenzoyl)-Z-aminophenyl-
`
`0.1 g
`
`acetate monohydrate
`Boric acid
`Borax
`5 Sodium chloride
`Methylparaben
`Ethylparaben
`Polyvinyl pyrrolidone
`Sodium sul?te
`Sterile puri?ed water
`10 PH 7-5
`
`TABLE 1
`
`1.0 t 0.4 (60.0)b
`1.3 i 0.2 *2(65.8)b
`
`0.1 i 0.1 (85.7)’?
`
`4.5 i 0.6 (30.8)”
`
`0 s1 (100)b
`6.8 i 1.0 *1(50.4)1>
`
`Test Drug
`Physiological Compound [I]
`saline (14)“ 0.1% (8)“
`Parameter
`m
`Corneal opacity
`2.5 i 0.5
`Palpebral cunjunctival
`3.8 i 0.5
`injection
`Palpebral conjunctiva]
`edema
`Balbar conjunctival
`injection
`Discharge
`Total Score
`Mam
`Anterior chamber
`opacity
`lridic injection
`Morphological
`change of iris
`Total Score
`External + lnternal
`Grand Total Score
`
`3.6 i 0.8 (_20.0)b
`
`2.6 r 0.2 “(57.4%
`2.9 i 0.4 t2(34.1)l7
`
`13.5 i‘ 0.8
`
`9.1 i 1.0 *2(32.6)b
`
`27.3 i 2.5
`
`0.1 g
`Sufficient quantity
`0.8 g
`0.3 g
`0.1 g
`2.0 g
`0.1 g
`To make 100 ml
`
`Compound [I]
`0.5% (10)"
`
`O #1000“?
`
`