JP2006348023A - Vascularization inhibitor comprising amine derivative as effective ingredient - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a vascularization inhibitor as a preventive or therapeutic agent particularly used for ophthalmic diseases accompanying vascularization such as diabetic retinopathy or macular degeneration. <P>SOLUTION: The vascularization inhibitor comprises as an effective ingredient a compound represented by general formula [I] (wherein the ring A is a benzene ring or a naphthalene ring; X is a single bond, an alkylene group or -CH<SB>2</SB>COOCH<SB>2</SB>-: Y is an oxygen atom or N(R<SB>6</SB>); R<SB>1</SB>, R<SB>2</SB>and R<SB>3</SB>are the same or different and are each a hydrogen atom or an alkyl group, and R<SB>1</SB>and R<SB>3</SB>can be bonded together to form a ring having one or a plurality of double bonds; R<SB>4</SB>and R<SB>5</SB>are the same or different and are each a hydrogen atom or an alkyl group; and R<SB>6</SB>is a hydrogen atom or an alkyl group) or a salt thereof. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

The present invention is an angiogenesis inhibitor comprising a compound represented by the general formula [I] or a salt thereof as an active ingredient, and in particular, a prophylactic or therapeutic agent for ocular diseases associated with angiogenesis such as diabetic retinopathy and macular degeneration. About.

  Vascular homeostasis is maintained by various functions of endothelial cells. Vascular endothelial cells 1) mediate the transport of necessary components such as nutrients in the blood to the tissue and prevent unnecessary large amounts of components from passing through. 2) Smooth circulation without clotting of blood. 3) an action to prevent bleeding when the blood vessel is disconnected, and 4) a regulating action to keep the blood vessel tension constant.

Angiogenesis occurs at the stage of degradation of the basement membrane by proteases produced by vascular endothelial cells, migration / proliferation of vascular endothelial cells, formation of lumens of vascular endothelial cells, formation of basement membranes and pericytes. Angiogenesis is closely linked to various diseases, especially diabetic retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein occlusion, retinal artery occlusion, pterygium, rubeosis, pannus, etc. There is.
On the other hand, Patent Document 1, Patent Document 2 and Patent Document 3 describe that the compound represented by the general formula [I] or a salt thereof has trypsin inhibitory activity and is effective in the treatment of pancreatitis. However, it has not been studied at all what pharmacological action the compound exhibits for eye diseases such as diabetic retinopathy and macular degeneration with angiogenesis.
Japanese Patent Laid-Open No. 57-053454 JP-A-61-33173 JP 51-138642 A

  It is a very interesting subject to search for new pharmaceutical uses for the compound represented by the general formula [I] or salts thereof.

  In order to find out a new pharmacological action of the compound represented by the general formula [I] or a salt thereof, the present inventors examined the action of the compound on angiogenesis. As a result, it has been found that the compound has an excellent angiogenesis inhibitory action and is useful as a prophylactic or therapeutic agent for ocular diseases associated with angiogenesis.

That is, the present invention is an angiogenesis inhibitor comprising a compound represented by the general formula [I] or a salt thereof (hereinafter collectively referred to as “the present compound”) as an active ingredient. It is useful as a preventive or therapeutic agent for diabetic retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein occlusion, retinal artery occlusion, pterygium, rubeosis, pannus and the like.

[Wherein, ring A represents a benzene ring or a naphthalene ring, X represents a single bond, an alkylene group or —CH ₂ COOCH ₂ —, Y represents an oxygen atom or N (R ₆ ), and R ₁ , R ₂ And R ₃ may be the same or different and each represents a hydrogen atom or an alkyl group, or R ₁ and R ₃ may be bonded to form a ring having one or more double bonds, and R ₄ and R ₅ are the same Or, differently, it represents a hydrogen atom or an alkyl group, R ₆ represents a hydrogen atom or an alkyl group. ]
The present invention also provides an angiogenesis inhibitor comprising a compound represented by the general formula [II] or a salt thereof as an active ingredient, and the compound is diabetic retinopathy, retinopathy of prematurity, macular degeneration, angiogenesis. It is useful as a preventive or therapeutic agent for glaucoma, retinal vein occlusion, retinal artery occlusion, pterygium, rubeosis, pannus and the like.

[Where:
R ₁ , R ₂ and R ₃ may be the same or different and each represents a hydrogen atom or an alkyl group, or R ₁ and R ₃ may be bonded to form a ring having one or more double bonds. ]
Preferred examples of this compound include 6′-amidino-2′-naphthyl-4-guanidinobenzoate dimethanesulfonate (hereinafter referred to as “nafamostat”) represented by the following formula [Ia], and the following formula [Ib]. ] 6'-amidino-2'-naphthyl- (4- (4,5-dihydro-1H-2-imidazolyl) amino) benzoate dimethanesulfonate (hereinafter referred to as "FUT-187") and N, N-dimethylcarbamoylmethyl-4- (4-guanidinobenzoyloxy) phenyl acetate methanesulfonate (hereinafter referred to as “camostat”) represented by the following formula [Ic] is exemplified.

  This compound can be produced according to a usual method in the field of synthetic organic chemistry, and detailed production methods are described in Patent Documents 2 and 3.

  Here, each group and wording prescribed | regulated in this specification are shown below. “Alkylene” refers to a linear or branched alkylene having 1 to 6 carbon atoms, and specific examples include methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, methylmethylene, dimethylmethylene, propylene, Examples include 2-methyltrimethylene. “Alkyl” means straight or branched alkyl having 1 to 6 carbon atoms, and specific examples include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl. , Sec-butyl, tert-butyl, isopentyl and the like. In addition, “may form a ring having one or more double bonds” means 2-imidazolyl, 4,5-dihydro-1H-2-imidazolyl, 2-pyrimidyl, 1,4,5,6 -It means that a ring such as tetrahydro-2-pyrimidyl may be formed.

  The present compounds can take the form of “prodrugs”. The term “prodrug” as used herein means a compound obtained by modifying the present compound with a group capable of leaving in vivo. In particular, a group containing an acylatable nitrogen atom such as an amino group or a guanidino group is acylated or formed. Mention may be made of prodrugs in the form of carbamate derivatives.

  The salt of this compound may be any pharmaceutically acceptable organic or inorganic acid addition salt, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, acetic acid, phosphoric acid, lactic acid, citric acid, Examples thereof include organic acids such as methanesulfonic acid, fumaric acid, maleic acid, mandelic acid, citric acid, tartaric acid, and salicylic acid.

  This compound has optical isomers and, in some cases, diastereomeric isomers, and those containing these as active ingredients are also included in the present invention. Further, the present compound may take the form of a solvate, for example, a hydrate.

  The angiogenesis inhibitory action of the present compound is based on the fact that the compound represented by the above general formula [I] or a salt thereof has an excellent angiogenesis inhibitory action in a pharmacological test for rat laser-induced choroidal neovascularization described later. is there.

  The compound can be formulated as necessary by adding a pharmaceutically acceptable additive and using a technique widely used as a single preparation or a combined preparation.

  When this compound is used for the prevention or treatment of the aforementioned eye diseases, it can be administered to patients orally or parenterally. As the dosage form, oral administration, topical administration to the eye (instillation administration) , Intravitreal administration, subconjunctival administration, subtenon administration, etc.), intravenous administration, transdermal administration, etc., and agents suitable for administration together with pharmaceutically acceptable additives as necessary Formulated into molds. Examples of dosage forms suitable for oral administration include tablets, capsules, granules, powders, etc. Examples of dosage forms suitable for parenteral administration include injections, eye drops, eye ointments, gels, Examples include nasal drops and suppositories. These can be prepared using conventional techniques widely used in the field. In addition to these preparations, this compound can also be made into preparations for DDS (drug delivery system) such as preparations for intraocular implants and microspheres.

  For example, the tablet is an excipient such as lactose, glucose, D-mannitol, anhydrous calcium hydrogen phosphate, starch, sucrose; carboxymethylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, crospovidone, starch, partially pregelatinized starch , Disintegrating agents such as low-substituted hydroxypropylcellulose; binders such as hydroxypropylcellulose, ethylcellulose, gum arabic, starch, partially pregelatinized starch, polyvinylpyrrolidone, polyvinyl alcohol; magnesium stearate, calcium stearate, talc, hydrous silicon dioxide , Lubricants such as hydrogenated oils; refined sucrose, hydroxypropylmethylcellulose, hydroxypropylcellulose, methylcellulose, ethylcellulose, polyvinyl chloride Coating agents pyrrolidone; citric acid, aspartame, using ascorbic acid, appropriately selected and the like flavoring agent such as menthol, can be prepared.

  Injections can be prepared using sterile purified water and sodium chloride for isotonicity.

  Eye drops include isotonic agents such as sodium chloride and concentrated glycerin; buffering agents such as sodium phosphate and sodium acetate; surface activity such as polyoxyethylene sorbitan monooleate, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil Agents; Stabilizers such as sodium citrate and sodium edetate; Preservatives such as benzalkonium chloride and paraben can be selected and used as necessary, and pH is acceptable for ophthalmic preparations Although it may be in the range, it is usually preferably in the range of 4-8.

  The preparation for intraocular implant can be prepared using a biodegradable polymer, for example, a biodegradable polymer such as polylactic acid, polyglycolic acid, lactic acid / glycolic acid copolymer, and hydroxypropylcellulose.

  The dose of this compound can be appropriately changed depending on the dosage form, the severity of the patient's symptoms, age, weight, doctor's judgment, etc. 0.01-2000 mg per day can be administered once or several times. In the case of injections, generally 0.01-200 mg per day is divided into one or several times for an adult. In the case of eye drops, the active ingredient concentration of 0.01 to 10% (w / v) is usually instilled once or several times a day. Furthermore, in the case of an intraocular implant preparation, an intraocular implant preparation containing 0.01 to 2000 mg for an adult can be implanted in the eye.

  As will be described in detail in the section of the pharmacological test described later, when the action of the compound represented by the above general formula [I] or a salt thereof on rat laser-induced choroidal neovascularization was examined, it was represented by the above general formula [I]. The compounds or salts thereof showed a strong inhibitory effect on choroidal neovascularization. Accordingly, the compound represented by the above general formula [I] or a salt thereof of the present invention is an angiogenesis inhibitor, retinopathy of retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein. It turns out that it is useful as a preventive or therapeutic agent for obstruction, retinal artery occlusion, pterygium, rubeosis, pannus and the like. A similar test was performed for a known compound similar to the present compound, but the present compound showed a much higher effect than the known compound, confirming the excellent usefulness of the present compound.

  Hereinafter, the present invention will be described in detail with reference to pharmacological tests and formulation examples, but these are for better understanding of the present invention and do not limit the scope of the present invention.

[Pharmacological test]
In order to investigate the angiogenesis inhibitory action of the compound represented by the general formula [I] or salts thereof, rat laser induction of the compound represented by the general formula [I] or salts thereof, nafamostat, FUT-187 and camostat The effect on choroidal neovascularization was examined. As a comparative compound, ethyl-4- (6-guadininohexanoyloxy) benzoate methanesulfonate (hereinafter referred to as “gabexate”) represented by the following formula [III] was used.

  Comparing the chemical structures of gabexate with nafamostat, FUT-187 and camostat, they are common in that they have a guanidino group and an aromatic ring, but others have different chemical structures. In addition, gabexate is known to have trypsin inhibitory activity and to be effective in the treatment of pancreatitis, and as described in the background section, nafamostat, FUT-187 and camostat have the same activity. Thus, gabexate is common to the above three compounds according to the present invention.

(Production of rat laser-induced choroidal neovascularization model)
Rats were anesthetized by intramuscular administration of 1 ml / kg of a mixed solution (7: 1) of 5% (W / V) ketamine hydrochloride injection and 2% xylazine hydrochloride injection, and 0.5% (W / V) After tropicamide-0.5% phenylephrine hydrochloride ophthalmic solution was instilled to make mydriatic, photocoagulation was performed with a krypton laser photocoagulator. Photocoagulation was performed in the posterior region of the fundus, avoiding thick retinal blood vessels, and focusing on the deep retina at 8 spots per eye (coagulation conditions: spot size 100 μm, output 100 mW, coagulation time 0.1 seconds). . After photocoagulation, fundus photography was performed to confirm the laser irradiation site.

(Drug administration method and evaluation method)
1-1. Intraperitoneal administration Nafamostat was dissolved in 5% (W / V) glucose solution (prepared by dissolving D-glucose in physiological saline) to 25 mg / ml, and the nafamostat solution was administered at a dose of 1 ml / kg. The abdominal cavity was administered once a day for 7 days from the photocoagulation surgery day, including the surgery day. Similarly, 25 mg / ml and 125 mg / ml FUT-187 solutions and 125 mg / ml camostat solution were prepared and intraperitoneally at a dose of 1 ml / kg for 1 day for 7 days from the photocoagulation operation day to the operation day. One dose was administered. In the comparative compound administration group, a solution in which gabexate was dissolved in a glucose solution to 50 mg / ml was administered in the same manner as described above. A 5% (W / V) glucose solution was similarly administered to the base administration group.

1-2. Evaluation Method On the 7th day after photocoagulation, 0.1 ml of a 10% fluorescein solution was injected from the tail vein to perform fluorescence fundus imaging. In fluorescence fundus angiography, a spot where no fluorescent exposure was observed was judged negative (no angiogenesis), and a spot where fluorescent exposure was seen was judged positive. In addition, a photocoagulation site where slight fluorescence exposure was observed was determined to be positive (with angiogenesis) when two sites were present. The new blood vessel appearance rate was calculated according to Equation 1. The neovascularization rate was calculated from the number of positive spots with respect to 8 spots irradiated with laser, and the choroidal neovascularization inhibition rate was calculated according to Equation 2. The results are shown in Table 1. The number of cases in each administration group is 4-7.

2-1. Oral administration Nafamostat and FUT-187 were dissolved in 1% (W / V) methylcellulose solution to a concentration of 20 mg / ml, and were administered at a dose of 5 ml / kg for 7 days from the day of photocoagulation surgery to the day of surgery. Orally administered 3 times a day. Similarly, camostat was suspended in 1% (W / V) methylcellulose solution at 20 mg / ml, and orally administered at a dose of 5 ml / kg three times a day for 7 days from the photocoagulation surgery day to the day of surgery. . A 1% (W / V) methylcellulose solution was similarly administered to the base administration group.

2-2. Evaluation method The evaluation method was performed in the same manner as described above. The results are shown in Table 2. The number of cases in each administration group is 6-7.

3-1. Intravitreal administration Nafamostat was dissolved in distilled water to 2 mM to obtain a nafamostat solution. Immediately after photocoagulation and on the third day after photocoagulation, rats were intramuscularly administered with 1 ml / kg of a mixture of 5% (W / V) ketamine hydrochloride injection and 2% xylazine hydrochloride injection (7: 1). After anesthesia, 0.5% (W / V) tropicamide-0.5% phenylephrine hydrochloride ophthalmic solution was instilled and mydriatic, then, using a 32G microsyringe, 5 μl of nafamostat solution was intravitreally under a surgical microscope. Was administered. Distilled water was similarly administered to the base administration group.

3-2. Evaluation method The evaluation method was performed in the same manner as described above. The results are shown in Table 3. The number of cases in each administration group is 5-6.

4-1. Subconjunctival administration Nafamostat was dissolved in a base (distilled water or 5% (W / V) glucose solution (prepared by dissolving D-glucose in physiological saline)) to 2 mg / ml, and the nafamostat solution Thereafter, 50 μl of nafamostat solution was administered subconjunctivally once a day for 7 days from the photocoagulation surgery day to the surgery day. A base (distilled water or 5% (W / V) glucose solution) was similarly administered to the base administration group. Optionally, before subconjunctival administration, rats were intramuscularly administered with 0.25 ml / kg of a mixture of 5% (W / V) ketamine hydrochloride injection and 2% xylazine hydrochloride injection (7: 1). Anesthetized.

4-2. Evaluation method The evaluation method was performed in the same manner as described above. The results are shown in Table 4. The number of cases in each administration group is 5-8.

Formula 1
Neovascular expression rate (%) = (number of positive spots / total number of photocoagulation sites) × 100
Formula 2
Choroidal neovascularization inhibition rate (%) = (A ₀ −A _X ) / A ₀ × 100
A ₀ : New blood vessel expression rate in the base administration group A _X : New blood vessel expression rate in the drug administration group

(Results and Discussion)
As is clear from the results in Table 1, nafamostat, FUT-187 and camostat all showed a high inhibition rate of choroidal neovascularization. On the other hand, gabexate did not show choroidal neovascularization inhibitory action. That is, it was found that the compound represented by the general formula [I] represented by nafamostat, FUT-187 and camostat or salts thereof showed an excellent choroidal neovascularization inhibitory action. Further, as is clear from the results of Tables 2 to 4, it was found that nafamostat or FUT-187 has an excellent choroidal neovascularization inhibitory effect also in oral administration, intravitreal administration and subconjunctival administration.

  Next, the pharmaceutical agent of the present invention will be described more specifically with reference to formulation examples, but the present invention is not limited to these formulation examples.

[Formulation example]
Formulation Example 1 Eye drops (in 1 ml)
Nafamostat 1mg
Concentrated glycerin 250mg
Polysorbate 80 200mg
Sodium dihydrogen phosphate dihydrate 20mg
1N sodium hydroxide appropriate amount 1N hydrochloric acid appropriate amount sterilized purified water appropriate amount Nafamostat and other ingredients mentioned above are added to sterilized purified water, and these are mixed well to make eye drops.

Formulation Example 2 Tablet (in 100 mg)
FUT-187 1mg
Lactose 66.4mg
Maize starch 20mg
Carboxymethylcellulose calcium 6mg
Hydroxypropylcellulose 6mg
Magnesium stearate 0.6mg
FUT-187 and lactose are mixed in a mixer, carboxymethylcellulose calcium and hydroxypropylcellulose are added thereto, this mixture is granulated, the resulting granules are dried and sized, and stearic acid is added to the sized granules. Magnesium is added and mixed, and the mixture is tableted with a tableting machine.

Formulation Example 3 Injection (in 10 ml)
FUT-187 10mg
Sodium chloride 90mg
Sterilized purified water appropriate amount FUT-187 and sodium chloride are dissolved in sterile purified water to make an injection solution.

A desired formulation can be obtained by appropriately changing the types and amounts of the present compound and additives including the above-mentioned formulations.

Claims (7)

An angiogenesis inhibitor comprising a compound represented by the following general formula [I] or a salt thereof as an active ingredient.

[Where:
Ring A represents a benzene ring or a naphthalene ring,
X represents a single bond, an alkylene group or —CH ₂ COOCH ₂ —,
Y represents an oxygen atom or N (R ₆ );
R ₁ , R ₂ and R ₃ are the same or different and each represents a hydrogen atom or an alkyl group,
Alternatively, R ₁ and R ₃ may combine to form a ring having one or more double bonds,
R ₄ and R ₅ are the same or different and each represents a hydrogen atom or an alkyl group,
R ₆ represents a hydrogen atom or an alkyl group. ] An angiogenesis inhibitor comprising a compound represented by the following general formula [II] or a salt thereof as an active ingredient.

[Where:
R ₁ , R ₂ and R ₃ are the same or different and each represents a hydrogen atom or an alkyl group,
Alternatively, R ₁ and R ₃ may combine to form a ring having one or more double bonds. ] The compound represented by the general formula [I] is 6′-amidino-2′-naphthyl-4-guanidinobenzoate, 6′-amidino-2′-naphthyl- (4- (4,5-dihydro-1H-2- The angiogenesis inhibitor according to claim 1, which is imidazolyl) amino) benzoate or N, N-dimethylcarbamoylmethyl-4- (4-guanidinobenzoyloxy) phenyl acetate. A prophylactic or therapeutic agent for a disease associated with angiogenesis, comprising a compound represented by the following general formula [I] or a salt thereof as an active ingredient.

[Where:
Ring A represents a benzene ring or a naphthalene ring,
X represents a single bond, an alkylene group or —CH ₂ COOCH ₂ —,
Y represents an oxygen atom or N (R ₆ );
R ₁ , R ₂ and R ₃ are the same or different and each represents a hydrogen atom or an alkyl group,
Alternatively, R ₁ and R ₃ may combine to form a ring having one or more double bonds,
R ₄ and R ₅ are the same or different and each represents a hydrogen atom or an alkyl group,
R ₆ represents a hydrogen atom or an alkyl group. ] The preventive or therapeutic agent according to claim 4, wherein the disease accompanied with angiogenesis is an ophthalmic disease. The preventive or therapeutic agent according to claim 5, wherein the ophthalmic disease is diabetic retinopathy, retinopathy of prematurity, macular degeneration, neovascular glaucoma, retinal vein occlusion, retinal artery occlusion, pterygium, rubeosis or pannus. The angiogenesis inhibitor according to any one of claims 1 to 3, or the prevention or treatment according to any one of claims 4 to 6, wherein the dosage form is an oral preparation, an injection, an eye drop or a preparation for an intraocular implant. Agent.