JP7109035B2 - COMPOSITIONS FOR MAINTENANCE OF BULBS - Google Patents

Description

The present invention relates to a composition that does not suppress the proliferation of corneal epithelial cells and suppresses the proliferation of fibroblasts, and furthermore, to a pharmaceutical composition for maintaining the filtering bleb formed in glaucoma filtration surgery.

Glaucoma is a progressive disease in which retinal ganglion cells die, presenting with atrophy of the optic nerve and loss of visual field. Basically, at present, it is difficult to restore visual field once it has been lost, so it can cause blindness.

Increased intraocular pressure (IOP) is considered to be one factor in the onset of glaucoma. It is also believed that this enhancement may be caused by abnormalities in the filtration mechanism of the aqueous humor, the fluid that circulates in the eye. In a normal eye, aqueous humor is produced in a tissue called the ciliary body, passes behind the iris, reaches the anterior chamber, passes through the trabecular meshwork, is discharged from Schlemm's canal, and flows into blood vessels outside the eye. It circulates on a fixed route. This circulation of aqueous humor produces a substantially constant pressure in the eye, which maintains the shape of the eyeball (see upper part of FIG. 1). However, when the filtration mechanism in the trabecular meshwork becomes insufficient, the amount of aqueous humor in the anterior chamber increases, causing an increase in intraocular pressure, which changes the shape of the eyeball and eventually damages the optic nerve papilla. Pressure is applied to the location, and it is believed that the retinal ganglion cells die, resulting in the development of glaucoma.

Therefore, current glaucoma treatment initially involves drug therapy with intraocular pressure-lowering drugs and laser therapy for the purpose of lowering intraocular pressure, but if these effects are insufficient, glaucoma filtration surgery, etc. Surgery may also be performed.

Glaucoma filtration surgery is the gold standard for controlling intraocular pressure and provides an alternative route for the drainage of aqueous humor from the eye. In this surgical method, for example, the conjunctiva of the corneal limbus (so-called inner side of the eyelid) and the sclera (so-called white of the eye), and further the trabecular meshwork at the bifurcation of these are incised, and then the incised conjunctiva and By suturing the sclera, the aqueous humor in the anterior chamber can flow out from the gap between the incised sclera and under the conjunctiva, which in turn reduces the intraocular pressure (lower part of Fig. 1). reference).

Here, the gap between the conjunctiva and the sclera (conjunctival blister), which is formed under the conjunctiva and temporarily accumulates the aqueous humor flowing out through the wound outside the conjunctiva, is called a filtering bleb. , the maintenance of this filtering bleb is a major factor in the success of glaucoma filtration surgery. That is, in the filtering bleb, postoperative proliferation and activation of fibroblasts in the conjunctiva often lead to the formation of scar tissue, which inhibits outflow of aqueous humor (Non-Patent Documents 1 and 2). Therefore, in many cases, the intraocular pressure lowering effect of the surgery is maintained only for about 2 to 3 years.

In this regard, as described above, the formation of scarred tissue is mainly due to the proliferation of fibroblasts and the like and the accompanying collagen accumulation (Non-Patent Document 3). Alternatively, mitomycin C (MMC) is applied to the scleral incision (MMC combined trabeculectomy, Non-Patent Documents 4 and 5). However, in some cases, even if the concentration of MMC is increased, a sufficient effect cannot be obtained. For example, it has been reported that in patients with fibrosis risk factors (young age, aphakic eyes, angiogenesis, inflammation, etc.), MMC may not have sufficient effects on the formation of scar tissue. (Non-Patent Document 6). In addition, due to its high cytotoxicity, there is a high possibility that other tissues will also be damaged. It is a problem that it occurs later (Non-Patent Document 7).

Therefore, there is a need for a compound that inhibits the proliferation of fibroblasts in the conjunctiva and sclera without damaging other tissues (such as the cornea). However, the current situation is that a method for maintaining the filtering bleb using such a compound in glaucoma filtration surgery has not yet been established.

Muckley ED. et al., Optom Vis Sci. , 2004, Vol. 81, No. 7, pp. 499-504 Francis BA. et al., Arch Ophthalmol. , 2005, Vol. 123, No. 2, pp. 166-170 Esson DW. et al., Invest Ophthalmol Vis Sci. , 2004, Vol. 45, No. 2, pp. 485-491 Chen-Wu Chen, Trans Asia Pac Acad Ophthalmol. , 1983, Vol. 9, pp. 172-177 Matsuda T. et al., Jpn J Ophthalmol. , 1996, Vol. 40, No. 4, pp. 526-32 Skuta GL. et al., Surv Ophthalmol. , 1987, Vol. 32, No. 3, pp. 149-170 Kremer I. et al., Acta Ophthalmol. , 2012, Vol. 90, No. 3, pp. 271-276

The present invention has been made in view of the problems of the prior art, and makes it possible to suppress the proliferation of fibroblasts in the conjunctiva and sclera without damaging other tissues such as the cornea. The aim is to find compounds. Another object of the present invention is to provide a composition that uses such a compound to safely and long-lastingly maintain a filtering bleb formed by glaucoma filtration surgery.

In order to achieve the above object, the present inventors first investigated the proliferation of Tenon's capsule-derived fibroblasts between the sclera and conjunctiva using 1274 compounds known as existing drugs for various diseases. was screened for compounds exhibiting an inhibitory activity equal to or greater than that of MMC, and 69 compounds were selected. Furthermore, considering corneal epithelial damage, which is one of the complications that occur after trabeculectomy combined with MMC, compound 49, which has low toxicity (cytostatic activity) to human-derived corneal epithelial cells, is targeted for these compounds. seeds were selected.

In addition, as a result of evaluating apoptosis-inducing activity from the viewpoint that these compounds can suppress cell proliferation by a new mechanism of action that is different from existing filtering bleb maintenance drugs, 10 compounds have that activity. It became clear that

In addition, from the above 49 kinds of compounds, those with a narrow concentration range (effective range) showing growth inhibitory effects on Tenon's capsule fibroblasts were excluded, and from the viewpoint of avoiding the onset of steroid glaucoma, etc., steroid activity was added. Exclude the ones you have. In addition, from the viewpoint of concern about non-selective cytotoxicity, compounds corresponding to surfactants were also excluded, and compounds that were not marketed were also excluded because their systemic toxicity had not been clarified. As a result, 27 kinds of compounds including colchicine, amsacrine, mebendazole, mitoxantrone, niclosamide, doxorubicin, griseofulvin, thioridazine, triclosan, etc. inhibited proliferation of fibroblasts in conjunctiva and sclera, was successfully selected as a compound that would not damage the tissues of In particular, colchicine, amsacrine, mebendazole, mitoxantrone, niclosamide, and doxorubicin are compounds that exhibit an inhibitory activity equal to or greater than that of MMC against the proliferation of Tenon's capsule-derived fibroblasts. Since triclosan also has apoptosis-inducing activity on Tenon's capsule-derived fibroblasts, it is assumed that this activity inhibits the formation of scar tissue in the filtering bleb.

Next, the present inventors confirmed the efficacy of the compounds selected by the above in vitro screening in vivo using glaucoma filtration surgery model rabbits. Specifically, in glaucoma filtration surgery, as a result of adding amsacrine to the incisions of the sclera and conjunctiva in place of MMC, it was found that a higher intraocular pressure-lowering effect than MMC was stably sustained. In glaucoma filtration surgery, addition of MMC to the incisions of the sclera and conjunctiva followed by addition of colchicine to the eye suppresses postoperative intraocular pressure elevation that could not be suppressed by MMC administration alone. also found, and completed the present invention.

That is, the present invention relates to a composition that does not suppress the proliferation of corneal epithelial cells and suppresses the proliferation of fibroblasts, and furthermore, to a pharmaceutical composition that maintains the filtering bleb formed in glaucoma filtration surgery. , in more detail below.
<1> Colchicine, amsacrine, mebendazole, mitoxantrone, niclosamide, doxorubicin, griseofulvin, thioridazine, triclosan, risedronate, benidipine, sulfadimethoxine, ritodrine, enalapril, ketoprofen, folic acid, tranexamic acid, cefoxitin, bezafibrate, zolmitriptan, amantadine, β-carotene, capreomycin, carbamazepine, cefadroxil, diethylstilbestrol, and cilazapril, containing as an active ingredient at least one compound selected from the group consisting of corneal epithelial cell proliferation and fibroblasts composition for inhibiting the growth of
<2> The composition according to <1>, which is a pharmaceutical composition for maintaining a filtering bleb.
<3> The composition according to <2>, wherein the composition is added to at least one of the incised sclera and conjunctiva to form a filtering bleb.
<4> The composition according to <2>, wherein the composition is added to the eye after formation of the bleb.

According to the present invention, by suppressing the proliferation of fibroblasts, it is possible to maintain the filtering bleb formed by the glaucoma filtration surgery for a long time and keep the intraocular pressure low and constant. In particular, compared with MMC, which is currently mainly used in glaucoma filtration surgery, it is possible to maintain the filtering bleb for a longer period of time by exhibiting a fibroblast proliferation inhibitory effect equal to or greater than that of MMC. Furthermore, since at least the corneal tissue is not damaged, it is highly safe and the filtering bleb can be maintained. In addition, according to the present invention, administration to the conjunctiva or sclera that has been incised to form a filtering bleb during surgery not only exerts a maintenance effect, but also by adding it to the eye after surgery. In addition, the filter bleb is highly safe and can be maintained for a long time.

BRIEF DESCRIPTION OF THE DRAWINGS It is a figure which shows the outline|summary (in the figure, upper part) of the aqueous humor circulation pathway (intraocular pressure maintenance mechanism) in an eye, and the outline|summary (in the figure, lower part) of glaucoma filtration surgery. In the figure, arrows indicate the flow of aqueous humor. Fig. 12 is a graph showing the results of analyzing the growth inhibitory activity of existing drug 1274 compounds against Tenon's capsule fibroblasts by Alamar blue assay. The vertical axis in the figure shows the relative value when the fluorescence intensity of each compound detected by the Alamar Blue assay is set to 100 for MMC. Fig. 3 is a graph showing the results of analysis of the growth inhibitory activity against corneal epithelial cells by Alamar Blue assay for 69 compounds exhibiting growth inhibitory activity against Tenon's capsule fibroblasts. The vertical axis in the figure shows the relative value when the fluorescence intensity of each compound detected by the Alamar Blue assay is set to 100 in DMSO. Results of annexin V assay analysis of apoptosis-inducing activity on Tenon's capsule fibroblasts for 49 compounds that exhibit proliferation-inhibitory activity on Tenon's capsule fibroblasts but do not inhibit proliferation of corneal epithelial cells. indicates The vertical axis in the figure indicates the ratio (%) of cells in which apoptosis was induced in the cells subjected to the assay. Compounds with arrows indicate 10 compounds that were found to have apoptosis-inducing activity. 1 is a graph showing the cytostatic activity of compounds of the present invention (colchicine, amsacrine, mebendazole, mitoxantrone, niclosamide, and doxorubicin) against Tenon's capsule fibroblasts along with their concentrations. The vertical axis in the figure indicates the ratio to the number of cells when each compound was not added. The horizontal axis in the figure indicates the concentration (0.01 to 10 μM) of the compound of the present invention added to Tenon's capsule fibroblasts (the notation in the figure is the same as in FIGS. 6 to 9). 1 is a graph showing the cytostatic activity of compounds of the present invention (griseofulvin, thioridazine, triclosan, risedronate, and benidipine) against Tenon's capsule fibroblasts along with their concentrations. 1 is a graph showing the cytostatic activity against Tenon's capsule fibroblasts and the added concentration of the compounds of the present invention (sulfadimethoxine, ritodrine, enalapril, ketoprofen, folic acid). 1 is a graph showing the cytostatic activity of compounds of the present invention (tranexamic acid, cefoxitin, bezafibrate, zolmitriptan, amantadine) against Tenon's capsule fibroblasts along with their added concentrations. 1 is a graph showing the cytostatic activity against Tenon's capsule fibroblasts and the added concentration of the compounds of the present invention (β-carotene, capreomycin, carbamazepine, cefadroxil, diethylstilbestrol). Fig. 2 is a graph showing changes in intraocular pressure difference after surgery in glaucoma filtration surgery model rabbits. In the figure, the vertical axis indicates the difference between the intraocular pressure in the eye to be operated and that in the non-operated eye (intraocular pressure difference). The plot in the figure shows the average value of each administration group, and the attached bar represents the standard error. Asterisks (*) in the figure indicate significant differences from the control group (physiological saline-administered group) by Student's t-test (*P<0.05, **P<0.01). . A dagger in the figure indicates a significant difference from the control group by the Aspin-Welch test (two daggers, P<0.01). A double dagger in the figure indicates that a significant difference was observed from the MMC-administered group by Student's t-test (double dagger P<0.05). Fig. 10 is a graph showing changes in bleb (filter bleb) score after surgery in glaucoma filtration surgery model rabbits. The plot in the figure shows the average value of each administration group, and the attached bar represents the standard error. Asterisks (*) in the figure indicate significant differences from the control group (physiological saline-administered group) by Student's t-test (*P<0.05, **P<0.01). . Fig. 2 is a graph showing changes in intraocular pressure difference after surgery in glaucoma filtration surgery model rabbits. In the figure, the vertical axis indicates the difference between the intraocular pressure in the eye to be operated and that in the non-operated eye (intraocular pressure difference). The plot in the figure shows the average value of each administration group, and the attached bar represents the standard error. An asterisk (*) in the figure indicates that a significant difference was observed from the physiological saline-administered group by Student's t-test (** P<0.01). A dagger in the figure indicates that a significant difference was observed from the physiological saline-administered group by the Aspin-Welch test (two daggers, P<0.01). The double dagger (double dagger) in the figure indicates that a significant difference was observed with respect to the physiological saline administration group by Dunnett's multiple comparison test (one double dagger P < 0.05, two double daggers P<0.01). The chapters (sections) in the figure indicate that Steele's multiple comparison test showed a significant difference from the physiological saline-administered group (one chapter P<0.05). Parallel marks (parallel) in the figure indicate that a significant difference was observed with respect to the MMC-administered group by Student's t-test (one parallel mark P < 0.05, two parallel marks P < 0.01 ). Paragraphs in the figure indicate that a significant difference was observed from the MMC-administered group by the Aspin-Welch test (one paragraph P<0.05, two paragraphs P<0.05). 01). Fig. 10 is a graph showing changes in bleb (filter bleb) score after surgery in glaucoma filtration surgery model rabbits. The plot in the figure shows the average value of each administration group, and the attached bar represents the standard error. Asterisks (*) in the figure indicate significant differences from the control group (physiological saline-administered group) by Wilcoxon test (*P<0.05, **P<0.01). The dagger in the figure indicates that a significant difference was observed from the control group by Steele's multiple comparison test (one dagger P < 0.05, two dagger P < 0.01 ). The double dagger in the figure indicates that a significant difference was observed from the MMC administration group by the Wilcoxon test (1 double dagger P < 0.05, 2 double daggers (P<0.01).

<Composition for maintaining filter bleb>
As shown in Examples below, as a result of screening 1274 compounds, which are existing drugs, as compounds that have the activity of suppressing the proliferation of fibroblasts but do not suppress the proliferation of corneal epithelial cells, the following table Twenty-seven compounds, shown in 1-3, were identified. In addition, it has been clarified that administration of these compounds makes it possible to maintain the filtering bleb formed by glaucoma filtration surgery safely and for a long time, and to keep the intraocular pressure low.

Accordingly, the present invention provides colchicine, amsacrine, mebendazole, mitoxantrone, niclosamide, doxorubicin, griseofulvin, thioridazine, triclosan, risedronate, benidipine, sulfadimethoxine, ritodrine, enalapril, ketoprofen, folic acid, tranexamic acid, cefoxitin, bezafibrate, zolmitriptan. , amantadine, β-carotene, capreomycin, carbamazepine, cefadroxil, diethylstilbestrol and cilazapril, containing as an active ingredient at least one compound selected from the group consisting of, does not inhibit the proliferation of corneal epithelial cells, and A composition for inhibiting proliferation of fibroblasts is provided.

The composition of the present invention may contain at least one of the 27 compounds described above, but it has an activity of suppressing the proliferation of fibroblasts, and is used as an existing filter bleb maintenance drug. It preferably contains at least one compound selected from colchicine, amsacrine, mebendazole, mitoxantrone, niclosamide, and doxorubicin, from the viewpoint of having a higher activity than mitomycin C (MMC) presently used. From the viewpoint of having an apoptosis-inducing activity on cells, it preferably contains at least one compound selected from griseofulvin, thioridazine and triclosan.

The compounds such as colchicine according to the present invention (hereinafter also referred to as the compounds of the present invention) have the activity of suppressing proliferation of fibroblasts. Also included are analogues, pharmacologically acceptable salts, or solvates.

Derivatives and analogues of the compounds of the present invention can be obtained by those skilled in the art by appropriately utilizing known chemical reactions, replacing the functional groups in the compounds shown in Tables 1 to 3 with other functional groups, or can be prepared by further introducing or the like.

Whether or not the derivatives and analogues thus prepared have the activity of suppressing the proliferation of cells (fibroblasts or corneal epithelial cells) can be determined, for example, by suppressing the proliferation of target cells. Measured in the presence and absence of, and if the former measured value (cell number, etc.) is lower than that of the latter, it can be determined to have an activity to suppress the growth of the cell. If the former measurement value is equivalent to that of the latter, it can be determined that the substance does not have growth-suppressing activity on the cell. Such measurement of cell proliferation is not particularly limited, and a person skilled in the art can appropriately select a known technique (eg, Alamar Blue assay) and perform it. In addition, in the compound of the present invention, it is desirable that the activity for inhibiting the proliferation of fibroblasts is higher than that of MMC when targeting Tenon's capsule-derived fibroblasts.

The pharmacologically acceptable salt is not particularly limited and can be appropriately selected according to the structure of each compound. , nitrates, sulfates, phosphates, etc.), organic acid salts (acetates, trifluoroacetates, benzoates, oxalates, malonates, succinates, maleates, fumarates, tartrates , citrate, methanesulfonate, ethanesulfonate, trifluoromethanesulfonate, benzenesulfonate, p-toluenesulfonate, glutamate, aspartate, etc.). Moreover, the solvate is not particularly limited and can be appropriately selected depending on the intended purpose. Examples thereof include hydrates and ethanolates.

In addition, the compounds of the present invention include all isomers and isomer mixtures such as tautomers, geometric isomers, optical isomers based on an asymmetric carbon, and stereoisomers. Furthermore, the compounds of the present invention undergo metabolism such as oxidation, reduction, hydrolysis, amination, deamination, hydroxylation, phosphorylation, dehydration-oxidation, alkylation, dealkylation, conjugation, etc. in vivo to produce the desired Compounds exhibiting activity are also included, and the present invention also includes compounds that undergo metabolism such as oxidation, reduction, hydrolysis, etc. in vivo to produce the compounds of the present invention (so-called prodrug forms).

In addition, the content of the compound of the present invention in the composition of the present invention is appropriately adjusted according to the type of the compound used, using the proliferation of fibroblasts and/or corneal epithelial cells as an index. 0.005 to 50% (w/v) is preferred, 0.001 to 30% (w/v) is more preferred, and 0.01 to 10% (w/v) is even more preferred. More specifically, the content of colchicine in the composition of the present invention is preferably 0.005 to 0.01% (w/v), and the content of amsacrine in the composition of the present invention is preferably 0.01% (w/v). 1-10% (w/v) is preferred.

The composition of the present invention is mainly in the form of a pharmaceutical composition, but can also be in the form of a reagent used for research purposes (eg, in vitro or in vivo experiments).

More specifically, the pharmaceutical composition of the present invention is as follows.

Colchicine, amsacrine, mebendazole, mitoxantrone, niclosamide, doxorubicin, griseofulvin, thioridazine, triclosan, risedronate, benidipine, sulfadimethoxine, ritodrine, enalapril, ketoprofen, folic acid, tranexamic acid, cefoxitin, bezafibrate, zolmitriptan, amantadine, beta-carotene , capreomycin, carbamazepine, cefadroxil, diethylstilbestrol and cilazapril, comprising as an active ingredient at least one compound for maintaining a bleb.

In the present invention, the term "filtering bleb", also called a bleb, means a conjunctival blister formed under the conjunctiva during glaucoma filtration surgery and formed by aqueous humor flowing out through a wound outside the eye under the conjunctiva. (see Figure 1). Maintenance of the bleb means a state in which the bleb formed under the conjunctiva in glaucoma filtration surgery is not blocked and the function of allowing aqueous humor to flow from the inside of the eye to the outside of the eye is maintained, more specifically. means that the intraocular pressure after glaucoma filtration surgery is maintained at a level lower than that before surgery due to aqueous humor drainage from the filtering bleb, and in humans, the intraocular pressure is preferably maintained at 12 mmHg or less More preferably, it is a state maintained at 10 mmHg or less.

In addition, the term "glaucoma filtration surgery" in the present invention means surgery to form a filtering bleb by incising at least one of the conjunctiva and the sclera. ctomy), non-perforating trabeculectomy, full-thickness filtration surgery, and surgery using implants (Express, etc.).

Examples of glaucoma to be treated for such surgery include primary open-angle glaucoma, secondary open-angle glaucoma, hyperaqueous production glaucoma, primary angle-closure glaucoma, secondary angle-closure glaucoma, plateau iris glaucoma, and mixed type. Glaucoma, developmental glaucoma, steroid glaucoma, exfoliation glaucoma, amyloid glaucoma, neovascular glaucoma, malignant glaucoma, capsular glaucoma of the lens.

In addition to the compounds of the present invention described above, the pharmaceutical composition of the present invention may optionally contain additives such as buffers, tonicity agents, stabilizers, preservatives, Antioxidants, high molecular weight polymers and the like can be added.

Examples of buffers include phosphoric acid or salts thereof, boric acid or salts thereof, citric acid or salts thereof, acetic acid or salts thereof, carbonic acid or salts thereof, tartaric acid or salts thereof, ε-aminocaproic acid, trometamol and the like. . From the viewpoint of buffering capacity in the weakly acidic region, boric acid or salts thereof, citric acid or salts thereof, acetic acid or salts thereof are preferred, and citric acid or salts thereof are particularly preferred. Phosphates include sodium phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate and the like. , sodium borate, potassium borate and the like, citrates include sodium citrate, disodium citrate, trisodium citrate and the like, and acetates include sodium acetate, potassium acetate and the like. Carbonates include sodium carbonate, sodium hydrogencarbonate and the like, and tartrates include sodium tartrate, potassium tartrate and the like. When the pharmaceutical composition of the present invention contains a buffering agent, the content of the buffering agent can be appropriately adjusted depending on the type of the buffering agent and the like. 0.01-5% (w/v) is more preferred, 0.1-3% (w/v) is more preferred, and 0.2-2% (w/v) is most preferred.

Further, the pH of the pharmaceutical composition of the present invention adjusted by blending a buffer is preferably 4.0 to 8.0, more preferably 4.5 to 7.5, and 5.0 to 7.0. More preferably, 5.5 to 6.5 is most preferable.

Examples of tonicity agents include ionic tonicity agents and nonionic tonicity agents. Ionic isotonizing agents include sodium chloride, potassium chloride, calcium chloride, magnesium chloride and the like, and nonionic isotonizing agents include glycerin, propylene glycol, sorbitol, mannitol and the like. When the pharmaceutical composition of the present invention is blended with a tonicity agent, the content of the tonicity agent can be appropriately adjusted depending on the type of the tonicity agent. v) is preferably 0.02 to 7% (w/v), more preferably 0.1 to 5% (w/v), particularly preferably 0.5 to 4% (w/v), 0.8-3% (w/v) is most preferred.

Examples of stabilizers include sodium citrate and the like. When a stabilizer is added to the pharmaceutical composition of the present invention, the content of the stabilizer can be appropriately adjusted depending on the type of stabilizer.

Examples of preservatives include benzalkonium chloride, benzalkonium bromide, benzethonium chloride, sorbic acid, potassium sorbate, methyl parahydroxybenzoate, propyl parahydroxybenzoate, chlorobutanol and the like. When the pharmaceutical composition of the present invention contains a preservative, the content of the preservative can be appropriately adjusted depending on the type of preservative and the like. 0.0005-0.1% (w/v) is more preferred, 0.001-0.05% (w/v) is more preferred, and 0.002-0.01% (w/v) is most preferred.

Examples of antioxidants include ascorbic acid, tocopherol, dibutylhydroxytoluene, butylhydroxyanisole, sodium erythorbate, propyl gallate, sodium sulfite, and the like. The content of the antioxidant when the pharmaceutical composition of the present invention is blended with the antioxidant can be appropriately adjusted depending on the type of antioxidant, etc., but 0.0001 to 1% (w / v) is Preferably, 0.0005 to 0.1% (w/v) is more preferable, 0.001 to 0.02% (w/v) is more preferable, and 0.005 to 0.010% (w/v) is Most preferred.

A high molecular weight polymer can be blended as appropriate. Examples of high molecular weight polymers include methylcellulose, ethylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxyethylmethylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose acetate succinate, hydroxypropylmethylcellulose phthalate, Carboxymethylethyl cellulose, cellulose acetate phthalate, polyvinylpyrrolidone, polyvinyl alcohol, carboxyvinyl polymer, polyethylene glycol and the like. The content of the high-molecular-weight polymer when blending the high-molecular-weight polymer in the pharmaceutical composition of the present invention can be appropriately adjusted depending on the type of the high-molecular-weight polymer, etc. v) is preferred, 0.01 to 1% (w/v) is more preferred, and 0.1 to 0.5% (w/v) is even more preferred.

The dosage form of the pharmaceutical composition of the present invention is not particularly limited, but it is mainly administered parenterally. It may be added to at least one of the membrane and conjunctiva. For such addition, suitable dosage forms of the pharmaceutical composition of the present invention include eye drops, liquids, injections, and eye ointments. Also, as shown in the examples below, the pharmaceutical compositions of the invention may be added to the eye after formation of the bleb. In the case of such addition, suitable dosage forms of the pharmaceutical composition of the present invention include eye drops and liquids.

Eye drops, solutions or injections are prepared by dissolving the above-described compound of the present invention in an aqueous solvent (physiological saline, sterilized purified water, etc.) or non-aqueous solvent (vegetable oil, etc.), It can be prepared by appropriately adding stabilizers, preservatives, antioxidants and the like. The ophthalmic ointment is prepared by adding the above-mentioned high-molecular-weight polymer as an ointment base to the above-mentioned compound of the present invention, and optionally adding the above-mentioned buffer, tonicity agent, stabilizer, antiseptic and antiseptic. It can be prepared by appropriately adding an oxidizing agent or the like.

Compositions of the invention may contain, in addition to the compounds of the invention, one or more other compounds for maintaining the bleb. Such other compounds are not particularly limited, and commercially available or developing bleb maintenance agents are preferably used, more specifically, antimetabolites such as MMC and 5-fluorouracil (5-FU). is mentioned. The content of the compound in the pharmaceutical composition of the present invention can be appropriately adjusted depending on the type of compound, etc., but is preferably 0.001 to 50% (w/v), and 0.01 to 10% (w/ v) is more preferred. Moreover, when MMC is contained, it is most preferably 0.04% (w/v).

The composition of the present invention may also contain one or more glaucoma or ocular hypertension therapeutic agents or ocular hypotensive agents. The glaucoma therapeutic agent is not particularly limited, and for example, commercially available or under development glaucoma therapeutic agents are preferably used. Body blockers, β-receptor blockers, parasympathetic agents, carbonic anhydrase inhibitors, prostaglandins, and Rho kinase inhibitors. Specific examples of non-selective sympathomimetics include dipivefrin, specific examples of α2 receptor agonists include brimonidine and apraclonidine, and specific examples of α1 receptor blockers include bunazosin. specific examples of β-receptor blockers include timolol, befunolol, carteolol, nipradilol, betaxolol, levonolol, and metipranolol; specific examples of parasympathomimetics include pilocarpine; Specific examples of inhibitors include dorzolamide, brinzolamide, and acetazolamide, specific examples of prostaglandins include latanoprost, isopropyl unoprostone, bimatoprost, and travoprost, and specific examples of Rho kinase inhibitors includes Ripasudil.

The composition of the present invention can be stored in containers made of various materials. For example, a container made of polyethylene, polypropylene, or the like can be used, and from the viewpoints of ease of instillation (container hardness) and stability of the compound of the present invention, it is preferred to store in a polyethylene container. is preferred.

The product (medicine, reagent) of the composition of the present invention or its instructions may be labeled as being used to maintain a bleb. Here, "labeled on the product or instruction manual" means that the label is attached to the main body, container, packaging, etc. of the product, or instruction manuals, attached documents, advertising materials, and other printed materials that disclose product information It means that the display is attached to etc. In the indication that the composition of the present invention is used for suppressing the proliferation of fibroblasts without suppressing the proliferation of corneal epithelial cells, or for maintaining the filtering bleb, administration or the like of the composition of the present invention will suppress the filtering bleb. can include information about the mechanisms that maintain Mechanisms include, for example, information on suppression of scarring of filtering blebs by suppression of proliferation of fibroblasts. In addition, the label to the effect that it is used for maintaining a filtering bleb can include information about its use for treatment of glaucoma, or the like.

<Method for maintaining filter bleb>
As shown in the examples below, the bleb can be maintained by adding the compound of the present invention to a subject. Accordingly, the invention also provides a method of maintaining a bleb, comprising adding a compound or composition of the invention to a subject.

The compounds or compositions of the present invention can be used for animals including humans, but animals other than humans are not particularly limited, and various livestock, poultry, pets, experimental animals, etc. be able to. Specific examples include, but are not limited to, rabbits, monkeys, hamsters, mice, rats, pigs, cows, horses, sheep, goats, chickens, ducks, ostriches, ducks, dogs, and cats.

In one aspect of the method of the present invention, in glaucoma filtration surgery, the compound or composition of the present invention is added to at least one of the sclera and conjunctiva that has been incised to form a filtering bleb, Methods of maintaining a bleb are included.

The compound added in the method may be one compound selected from the above 27 compounds, or may be a combination of a plurality of compounds. etc.).

In addition, when the composition of the present invention is an eye drop or liquid, it may be applied dropwise to the incision during surgery, or soaked in absorbent paper, absorbent cotton, gauze, sponge, etc., to the incision. After temporary placement (usually about 5 minutes), the compound of the invention can be applied directly to the incision. When the composition of the present invention is an injection, it can be added by injecting it into the incision, and when it is an ophthalmic ointment, it can be added by applying it to the incision. Moreover, in such direct addition, the addition site is preferably the incision of both the sclera and the conjunctiva from the viewpoint of better maintenance of the postoperative filtering bleb.

Another embodiment of the method of the invention also includes a method of maintaining a bleb, comprising adding a compound or composition of the invention to the eye after the bleb has formed in glaucoma filtration surgery.

The compound added in the method may be one compound selected from the above 27 compounds, or may be a combination of a plurality of compounds, and other filter bleb maintenance agents (e.g. , MMC).

In addition, when the composition of the present invention is an eye drop or liquid, depending on the type and content of the compound of the present invention, once to several times a day (preferably 3 to 4 times a day) , drop by drop) can be added to the eye. In addition, although the timing of starting the addition depends on the state of healing after surgery, it is usually after 6 days have passed after the surgery. Also, it is desirable to add an antibacterial agent and/or an anti-inflammatory agent (eg, betamethasone sodium phosphate solution, levofloxacin hydrate solution) to the eye before and/or after the start of the addition. Also, sites of the eye to which such compounds are added include, for example, the corneal surface, subconjunctiva, and subtenon's capsule.

Furthermore, the method of adding such a composition of the present invention to the eye after surgery can be performed by existing glaucoma filtration surgery (for example, MMC-combined trabeculectomy), or by adding the above-mentioned compound of the present invention, etc. to the filtering bleb. When used in combination with glaucoma filtration surgery procedures that involve application to the incision to form, the filtering bleb can be maintained longer and the intraocular pressure can be kept lower.

EXAMPLES The present invention will be described in more detail based on examples below, but the present invention is not limited to the following examples.

<Primary screening using inhibition of fibroblast proliferation as an index>
In glaucoma filtration surgery, the formed filter bleb often becomes scarred tissue due to the proliferation of fibroblasts after the surgery, and outflow of aqueous humor is often inhibited. Therefore, at present, mitomycin C (MMC) is mainly applied (trabeculectomy combined with MMC) in order to suppress the proliferation of fibroblasts in the conjunctiva and sclera. However, there are cases where the suppression effect is not sufficient. Therefore, 1274 compounds known as existing drugs for various diseases were screened for compounds having higher fibroblast growth inhibitory activity than MMC.

The existing drug library used for screening was provided by Professor Hideyuki Saya of Keio University School of Medicine. In addition, Tenon's capsule fibroblasts (hereinafter also referred to as mTCF) derived from common marmosets, which are small primates closely related to humans, were used to evaluate the antiproliferative activity of these compounds. The method for preparing mTCF is as follows.

That is, first, Tenon's capsule was detached from the sclera of the common marmoset using scissors in Dulbecco's phosphate buffer and placed on a 10 cm dish supplemented with a minimum amount of Dulbecco's Modified Eagle's Medium (DMEM). In addition, DMEM (high glucose, manufactured by Life Technology) contains 1% antibiotics (penicillin and streptomycin, manufactured by Life Technology), and 10% fetal bovine serum (manufactured by Biological Industries) to prevent floating. is added. After transferring fibroblasts from the isolated tissue to a dish, explant culture was performed at 37° C. for 25 days. In addition, since the cells prepared in this way were FSP-1 positive by immunostaining for fibroblast-specific protein-1 (FSP-1) before being subjected to the following screening, It has been confirmed that they are all mTCF.

Then, the cytostatic activity against mTCF thus primary cultured was analyzed by Alamar Blue assay. Specifically, primary cultured mTCF was seeded in a 96-well plate at a cell density of 3.0×10 ³ cells/mm ² , and various existing drugs were added to a final concentration of 10 μM and cultured at 37° C. for 24 hours. did. The cells were then further cultured in the dark at 37° C. for 2 hours in DMEM supplemented with 10% Alamar Blue reagent (manufactured by Life Technology). Using a fluorescence microplate reader (manufactured by Molecular Devices, product name: Spectra Max Gemini), the intensity of fluorescence emitted from the cells to which each existing drug was added was measured at an excitation wavelength of 544 nm and an emission wavelength of 590 nm.

The absence of cells was defined as 0% control, and the presence of cells was defined as 100% control. As a result, the calculated CV value, S/B ratio, and Z' value were 5.96, 7.85, and 0.79, respectively, and all criteria (CV < 10, S/B > 2, Z' > 0 .5) was satisfied, the effectiveness of this system was confirmed, and drug screening was performed using it as described above.

As a result of this screening, as shown in FIG. 2, 67 kinds of compounds were successfully selected from among 1274 existing drugs, exhibiting growth inhibitory activity equal to or higher than that of MMC at the same concentration.

<Secondary screening using toxicity to corneal epithelial cells as an index>
Corneal epithelial damage is one of the complications that occur after trabeculectomy combined with MMC. Therefore, from the 67 compounds obtained by the primary screening, compounds with low toxicity to human-derived corneal epithelial cells, that is, compounds with low cytostatic activity against the cells were selected as follows. .

Human corneal epithelial cells used for evaluating the toxicity of these compounds were prepared as follows. That is, Hayashi R. The limbus of corneal tissue provided by the Northwest Lions Eye Bank was treated with 1.82 units/ml dispase II (Invitrogen company) containing DMEM medium, cultured at 37° C. for 1 hour, and further cultured in CnT-20 medium. Before being subjected to the following screening, immunostaining for human keratin 12 (K12) confirmed that the cells thus prepared were K12-positive, confirming that they were human corneal epithelial cells. I have

Then, the cytostatic activity against primary cultured human corneal epithelial cells was analyzed by Alamar blue assay. That is, 10 μM of each of the 67 compounds was added to human corneal epithelial cells seeded in a 96-well plate at a cell density of 2000 cells/mm ² . As a control group, a group to which only the solvent (DMSO) was administered was also prepared, and 24 hours later, cytotoxicity was evaluated using the Alamar Blue assay in the same manner as the above mTCF.

As a result, as shown in FIG. 3, 49 compounds, excluding 18 compounds that showed toxicity compared to the group administered only the solvent (DMSO), were found to have growth inhibitory activity against Tenon's capsule fibroblasts. while having a low toxicity (proliferation inhibitory activity) to corneal epithelial cells.

<Evaluation of fibroblast apoptosis-inducing activity>
For the purpose of finding a bleb-maintaining drug having a mechanism of action different from that of existing bleb-maintaining drugs, we focused on the apoptosis-inducing activity and performed screening by the method shown below.

In fact, there is also a report that inhibition of apoptosis in fibroblasts induces scar tissue formation during the wound healing process (Yoshikiyo Akasaka et al., "Expression of apoptosis-related antigens in protuberant and flat scars", Connective Tissue, 1998, 30 vol., pp. 29-35).

Therefore, the apoptosis-inducing activity of these compounds against mTCF was evaluated using the annexin V assay for the 49 compounds obtained by the above screening. That is, 10 μM of each of 49 compounds was administered to primary cultured mTCF, and 24 hours later, flow cytometric analysis was performed using annexin V labeled with a fluorescent dye. As a control, cells to which only DMSO was added were also prepared and analyzed in the same manner.

As a result, as shown in FIG. 4, 10 of the 49 compounds were found to have apoptosis-inducing activity.

<Tertiary screening using the effective range of drug efficacy, etc. as an index>
Next, with respect to 49 kinds of compounds selected in the secondary screening, the addition concentration was changed from 10 μM to 0.01 to 10 μM, and the cell growth inhibitory activity against mTCF was evaluated in the same manner as described above, and the concentration range showing its efficacy ( effective range) were determined (some of these results are shown in FIGS. 5-9). Compounds with a narrow effective range were then excluded.

In addition, substances with steroid activity are excluded from the viewpoint that they may cause an increase in intraocular pressure as an efficacy (side effect), and from the viewpoint of non-selective cytotoxicity, they are classified as surfactants. We also excluded compounds that Additionally, unmarketed compounds were also excluded in view of unknown systemic side effects.

As a result, we selected 27 compounds that are highly safe, such as inhibiting the proliferation of Tenon's capsule fibroblasts and having low toxicity to corneal epithelial cells, that is, compounds suitable for maintaining the filtering bleb. Selected compounds are shown in Tables 1-3.

Of the 9 compounds shown in Table 1, compounds 1 to 6 (colchicine, amsacrine, mebendazole, mitoxantrone, niclosamide, doxorubicin) have higher cytostatic activity against Tenon's capsule fibroblasts than MMC. is a compound. Compounds 7 to 9 (griseofulvin, thioridazine, and triclosan) were also found to have apoptosis-inducing activity on Tenon's capsule fibroblasts by the annexin V assay.

Next, the compound, which has a higher cytostatic activity against Tenon's capsule fibroblasts than MMC, was evaluated for efficacy in the in vivo system shown below.

<Establishment of trabeculectomy model>
First, a trabeculectomy model was established using white rabbits for in vivo drug efficacy evaluation. Specifically, a white rabbit was given general anesthesia by intramuscular injection of ketamine hydrochloride (40 mg/kg) and xylazine hydrochloride (4 mg/kg), and then the conjunctiva, sclera and The trabecular meshwork was dissected and a scleral flap was prepared. Then, about 100 to 200 μL of 0.4 mg/mL (0.04% (w/v)) MMC or physiological saline was soaked in water-absorbing paper (manufactured by Inami Co., Ltd., Medical quick absorber), subconjunctival and scleral flaps. Let sit on top for 5 minutes. The conjunctiva and sclera were sutured with 10-0 suture thread without suturing the scleral flap. Surgery was performed only on the right eye, and the left eye was left untreated. Also, no perioperative complications such as inadvertent penetration of the anterior chamber and filtering bleb were observed in any eye.

Up to 5 days after surgery, betamethasone sodium phosphate solution (manufactured by Shionogi & Co., Ltd., product name: Rinderon (registered trademark) 0.01%) and levofloxacin hydrate solution (manufactured by Santen Pharmaceutical Co., Ltd., product name: Cravit (registered trademark) ophthalmic solution 0.5%) was instilled into the eye three times a day. In addition, on the 1st, 3rd, 5th, 7th, 11th, 14th, 21st, 35th and 49th days after the operation, the intraocular pressure was measured and the filtering bleb was observed. In addition, all measurement days were carried out at the same time.

<Intraocular pressure measurement>
An applanation tonometer (manufactured by Reichert Technologies, product name: Model 30 Classic Pneumatonometer) is used to measure intraocular pressure, and the intraocular pressure of both eyes is measured 5 to 10 minutes after subjecting to general anesthesia in the same manner as above. did. For loss of corneal sensation, oxybuprocaine solution (contains 0.4% oxybuprocaine hydrochloride, manufactured by Santen Pharmaceutical Co., Ltd., Benoxil (registered trademark) ophthalmic solution) was topically added before the intraocular pressure measurement. . In addition, intraocular pressure was measured at 3-7 pm. Then, based on the obtained measurement results, the difference in intraocular pressure between the operated eye (right eye) and the non-operated eye (left eye) was calculated.

<Observation of filter bleb>
Filtration blebs were observed through a slit lamp and evaluated by a slightly modified method of Perkins et al. That is, evaluation was performed using the following scores that increase reflecting the height and size of the filtering bleb.
+0: No bleb is observed.
+1: Thickening is observed in the conjunctiva. No bleb is observed.
+2: Filtration bleb is observed in the range of 75 degrees.
+3: A bleb is observed in the range of 75 to 135 degrees.
+4: Filtration bleb is observed in the range of 135 degrees or more.

When the success or failure of the above surgery was evaluated using these two types of analysis methods, although not shown in the figure, intraocular pressure measurement showed the effect of maintaining intraocular pressure reduction by surgery and by MMC treatment. . Similarly, the observation of the bleb suggested the possibility that the bleb was formed by surgery and that the formed bleb lasted for a long time due to MMC treatment. Therefore, since the efficacy of MMC could be detected by analysis using the intraocular pressure or the filtering bleb as an indicator, the efficacy of the compound was evaluated in this evaluation system as follows.

<Evaluation of the effect of filtration surgery combined with MMC for maintaining intraocular pressure reduction>
Colchicine was evaluated as follows to determine whether or not postoperative eye drops in the currently performed MMC-combined filtration surgery can enhance the effect of maintaining the lowering of intraocular pressure of the surgery.

First, 20 rabbits were divided into 3 groups, 4 rabbits as a control group, 8 rabbits as an MMC-administered group, and 8 rabbits as an MMC and colchicine-administered group. In addition, the intraocular pressure was measured before the start of administration of colchicine, and it was confirmed that they were the same in the three groups.

In the control group, glaucoma filtration surgery was performed in the same manner as described above, except that water-absorbent paper containing about 100 to 200 μL of physiological saline was placed under the conjunctiva and on the scleral flap for 5 minutes. Physiological saline (50 μL) was instilled into the eyes three times a day from 6 to 14 days after the operation.

In addition, in the MMC-administered group, glaucoma filtration surgery was performed in the same manner as the control group, except that water-absorbent paper containing about 100 to 200 μL of 0.4 mg/mL MMC was placed under the conjunctiva and on the scleral flap for 5 minutes. Physiological saline was instilled into the eyes three times a day from 6 to 14 days after the operation.

In addition, in the MMC and colchicine administration group, glaucoma filtration surgery was performed in the same manner as in the MMC administration group, and colchicine (0.01% ( w/v), 50 μL) was instilled into the eyes one drop at a time.

In all groups, surgery was performed only on the right eye, and the left eye was left untreated. Also, no perioperative complications such as inadvertent penetration of the anterior chamber and filtering bleb were observed in any eye. Furthermore, the concentration of each compound to be added was set to a concentration that exhibits efficacy within a tolerable range of side effects. All groups were subjected to the above-described intraocular pressure measurement and filtering bleb observation on the 3rd, 7th, 14th, 21st, 28th, 35th, 42nd and 49th days after the operation. In addition, all measurement days were carried out at the same time. The results obtained are shown in FIGS.

As is clear from the results shown in FIG. 10, a decrease in intraocular pressure in the operated eye was observed in the 3 groups until 3 days after the operation compared with that in the non-operated eye. However, in the control group (physiological saline added group), the difference in intraocular pressure after that narrowed, and the difference almost disappeared after 14 days after the operation. In the MMC-administered group, the intraocular pressure-lowering effect of MMC was observed up to 35 days after the operation, but after that, the difference in intraocular pressure between the non-operated eye and the operated eye narrowed.

On the other hand, in the MMC and colchicine administration group, the attenuation of the intraocular pressure-lowering effect observed in the MMC administration group after 35 days after surgery was not observed. It was revealed that colchicine has an activity that enhances the intraocular pressure-lowering maintenance effect of the surgery.

In addition, as is clear from the results shown in FIG. 11, even in the bleb score analysis, colchicine eye drops after surgery can continue to suppress the disappearance of the formed filtering bleb, especially after 42 days after surgery. It became clear that cells could be maintained.

<Evaluation of intraocular pressure lowering maintenance effect as an intraoperative topical agent substituting for MMC>
In order to find a compound that has an effect of maintaining intraocular pressure lowering than MMC, which is used as a topical agent in current filtration surgery, it is useful as a drug to be applied intraoperatively to the incisions of the sclera and conjunctiva. , targeting amsacrine in said compound, and evaluated as follows.

First, 25 rabbits were divided into 5 groups, 5 rabbits as control group, 0.4 mg/mL (0.04% (w/v)), 5 rabbits as MMC administration group, 0.1% (w/v) /v), 1% (w/v) and 10% (w/v) amsacrine-administered groups of 5 rabbits each. In addition, intraocular pressure was measured before performing glaucoma filtration surgery, and it was confirmed that they were comparable in these groups.

In the control group, during the glaucoma filtration surgery, absorbent paper soaked with about 100 to 200 μL of physiological saline (Medical quick absorber manufactured by Inami Co., Ltd.) was placed under the conjunctiva and on the scleral flap for 5 minutes. The conjunctiva and sclera were sutured with 10-0 suture thread without suturing the scleral flap. Until 5 days after the operation, betamethasone sodium phosphate solution (manufactured by Shionogi & Co., Ltd., product name: Rinderon (registered trademark) 0.01%) and levofloxacin hydrate solution (manufactured by Santen Pharmaceutical Co., Ltd., product name : Cravit (registered trademark) ophthalmic solution 0.5%) was applied four times a day.

In addition, in the MMC-administered group, glaucoma filtration surgery was performed in the same manner as in the control group, except that approximately 100 to 200 μL of water-absorbent paper soaked with 0.4 mg/mL MMC was placed under the conjunctiva and on the scleral flap for 5 minutes. , Betamethasone sodium phosphate solution and levofloxacin hydrate solution were instilled four times a day until 5 days after the operation.

In the amsacrine-administered group, approximately 100 to 200 μL of water-absorbing paper soaked with 0.1% (w/v), 1% (w/v) or 10% (w/v) amsacrine was applied to the subconjunctival and scleral flaps. Glaucoma filtration surgery was performed in the same manner as the control group except that the subjects were left on the top for 5 minutes, and betamethasone sodium phosphate solution and levofloxacin hydrate solution were instilled four times a day until 5 days after the operation.

In all groups, surgery was performed only on the right eye, and the left eye was left untreated. Also, no perioperative complications such as inadvertent penetration of the anterior chamber and filtering bleb were observed in any eye. Furthermore, the concentration of each compound to be added was set to a concentration that exhibits efficacy within a tolerable range of side effects. For all groups, the above-described intraocular pressure measurement and filtering bleb observation were performed on days 7, 14, 21, 28, 42, 56, and 70 after surgery. In addition, all measurement days were carried out at the same time. The results obtained are shown in FIGS.

As is clear from the results shown in FIG. 12, a decrease in the intraocular pressure of the operated eye was observed in all groups up to 7 days after the operation compared with that of the non-operated eye. However, in the control group (physiological saline added group), the difference in intraocular pressure after that narrowed, and the difference almost disappeared after 21 days after the operation. In addition, in the MMC-administered group, the intraocular pressure-lowering effect of MMC was observed until 70 days after the operation, but after 28 days after the operation, the difference in intraocular pressure between the non-operated eye and the operated eye gradually decreased. A trend was observed.

On the other hand, in the amsacrine-administered group, when the administration concentration was 1%, the reduction in intraocular pressure and its sustained effect were observed as in the MMC-administered group. Furthermore, in the group administered with 10% (w/v) amsacrine, it was confirmed that the intraocular hypotensive effect lasted for a very long time from the 7th day after the operation until the 70th day when the measurement was completed.

In addition, as is clear from the results shown in FIG. 13, the bleb score analysis also revealed that the amsacrine administration maintained the filtering bleb for an extremely long period of time.

From the above results, it was revealed that amsacrine has a stronger intraocular pressure-lowering effect than the existing drug MMC.

As described above, according to the present invention, the proliferation of fibroblasts can be suppressed without suppressing the proliferation of corneal epithelial cells. By suppressing the proliferation of fibroblasts, it is possible to maintain the filtering bleb formed by the glaucoma filtration surgery for a long time and keep the intraocular pressure low and constant. In particular, compared with MMC, which is currently mainly used in glaucoma filtration surgery, it is possible to maintain the filtering bleb for a longer period of time by exhibiting a fibroblast proliferation inhibitory effect equal to or greater than that of MMC. Furthermore, since at least the corneal tissue is not damaged, it is highly safe and the filtering bleb can be maintained. In addition, according to the present invention, administration to the conjunctiva or sclera that has been incised to form a filtering bleb during surgery not only exerts the effect of maintaining it, but also by instillation after surgery, The filtering bleb is highly safe and can be maintained for a long time. Therefore, it is useful as a composition or the like for maintaining a filtering bleb administered during or after glaucoma filtration surgery.

Claims (3)

A pharmaceutical composition for maintaining a bleb, comprising amsacrine as an active ingredient. filter bleb added to the incised sclera and/or conjunctiva to form the1described inMedicineComposition. filter bleb is added to the eye after the formation of1described inMedicineComposition.

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