WO2001010442A1

WO2001010442A1 - Retinal ganglion cell death inhibitors containing dihydropyridine compounds

Info

Publication number: WO2001010442A1
Application number: PCT/JP2000/005321
Authority: WO
Inventors: Yasumasa Otori
Original assignee: Fujisawa Pharmaceutical Co., Ltd.
Priority date: 1999-08-09
Filing date: 2000-08-09
Publication date: 2001-02-15

Abstract

Retinal ganglion cell death inhibitors which contain as the active ingredient dihydropyridine compounds represented by general formula (I) or salts thereof wherein R1 represents nitrophenyl; and R?2, R3 and R4¿ represent each lower alkyl. Because of having an effect of inhibiting retinal ganglion cell death, the drugs containing the above-described dihydropyridine compounds (I) are expected as exerting efficacious and safe therapeutic effects on diseases in the field of ophthalmology in association with retinal ganglion cell death such as retinal degeneration, optic nerve disease and glaucoma.

Description

Description Retinal ganglion cell death inhibitor containing dihydroxypyridine compound

The present invention relates to a novel drug having an inhibitory effect on retinal ganglion cell death. Specifically, by exerting an inhibitory effect on retinal ganglion cell death, retinal degeneration, optic neuropathy (optic neuritis, papillitis, optic retinitis, etamptorol optic neuropathy, diabetic optic nerve accompanied by ganglion cell death) Disease, ischemic optic neuropathy, etc.) and glaucoma. Background art

General formula (I)

(Wherein, R ¹ represents nitrophenyl, and R ² , R ³ and R ⁴ each represent a lower alkyl group)

Dihydric Doropirijin compound represented in a two Rubajipin especially R 'is the 3-two Torofueniru R ² isopropyl, R ³ and R ⁴ are methyl is known as C a antagonist, activated by depolarization of the cell membrane It is known to block L-type voltage-gated calcium channels and suppress the influx of intracellular calcium, and is widely used clinically, for example, as a hypotensive agent. In addition, dihydropyridine compounds such as nilvadipine improve peripheral blood flow in eye tissues such as the optic disc, choroid, and retina, thereby improving blood flow in the normal eye. It has also been reported that it exerts an effective effect on the treatment of visual field narrowing, optic neuropathy, retinopathy, retinal degenerative disease, etc. associated with tension glaucoma (Japanese Patent Application No. 8-108284). However, what effect it has on retinal ganglion cell death; and what kind of therapeutic effect it has on ophthalmic diseases associated with retinal ganglion cell death is not yet known. Disclosure of the invention

The present inventors examined the pharmacological action of dihydropyridine compounds such as nilvadipine on retinal ganglion cell death and investigated dihydropyridine compounds such as nilvadipine, which are already known to be safe and have few side effects. We have been conducting research to explore new medical indications in ophthalmology and other fields.

As a result of examining the pharmacological effects of dihydropyridine compounds on retinal ganglion cell death, they were found to exert an excellent inhibitory effect. Therefore, it was revealed that the dihydropyridine compound can be expected to have an excellent clinical therapeutic effect in the treatment of ophthalmic diseases (eg, glaucoma, optic neuropathy, and retinal degeneration) associated with retinal ganglion cell death.

That is, the present invention provides a compound represented by the general formula (I):

A dihydropyridine compound represented by, among which 6-cyano-5-methoxy-2-carbonyl-2-methyl-3- (3-ditrophenyl) —1,4—dihydropyridine-13—hydrorubonic acid (generic name: nilvadipine) ) Or a retinal ganglion cell death inhibitor containing a salt thereof as an active ingredient. As a more specific clinical therapeutic agent, the dihydric pyridine compound represented by diluvadipine or a salt thereof is used as an active ingredient. The present invention provides a therapeutic agent for diseases associated with retinal ganglion cell death (ophthalmic diseases such as glaucoma, optic neuropathy, and retinal degeneration). You. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a graph showing the results of examining the survival rate of retinal ganglion cells by simultaneous administration of nilvadipine and glutamate. BEST MODE FOR CARRYING OUT THE INVENTION

Before describing the requirements that constitute the present invention, first, “retinal ganglion cell death”, which is the subject of the present invention, will be described.

Retinal ganglion cells are the only nerve cells in the retina that project to the brain, and death of these nerve cells is caused by ophthalmic diseases such as ocular or normotensive glaucoma, optic neuropathy, and retinal degeneration. It is known to be seen in the process of progress.

Therefore, a compound that exerts an inhibitory effect on retinal ganglion cell death can be expected to have an excellent therapeutic effect on the above-mentioned various diseases. In vitro basic experiments using nodal cells are in progress.

However, in order to perform cell death experiments using retinal ganglion cells, which are originally considered difficult to culture, it is preferable that the cultured cells themselves be morphologically and physiologically close to in vivo . The present inventors have studied a pure culture method of retinal ganglion cells that is as close as possible to in vivo and reported previously (“Ayumi of Medicine”, vol.189, No.4, P219-222, 1999). ).

This culture method uses retinal ganglion cells in a serum-free medium containing neurotrophic factors. In a pure culture, cells that survive for a long time have axons or neurites without exception. The present inventors performed calcinein staining, which is activated by endogenous esterase and emits fluorescence, and survives those that have axons or dendrites at least twice the major axis of the cell body Was considered. The present inventors focused on the fact that pure cultured retinal ganglion cells caused cell death by low concentration of daltamic acid (25 M), and used pure cultured rat retinal ganglion cells to produce glutamate. We examined the mechanism of neuronal cell death. As a result, glutamate neuronal death depends on the N-methyl-D-aspartate (NMDA) receptor antagonist APV (2-amino-5-phosphonovalerate) and L-type potential. A-amino-3-hydroxy-5-methylisoxazol-4-propionic acid-kainate (kainate) (AM PA—KA, non-NMDA) It can be suppressed by DNQX (6,7-dinitroquinoxaline-2,3-dione), a receptor antagonist. From the above results, it was clarified that low-concentration glutamate causes Caz ⁺ to flow into cells via the AMPA-KA receptor in cultured retinal ganglion cells, causing neuronal cell death.

Glutamate is an excitatory neurotransmitter abundantly present in the brain and retina, and plays an important role in the transmission of information in nerve tissue.On the other hand, an excessive increase in extracellular dartamic acid concentration causes neuronal death. It is also well known to bring. In recent years, it has been reported that the intravitreal glutamate concentration in glaucoma patients is about twice as high as in healthy subjects, suggesting that glutamate neuronal cell death may be involved in the pathogenesis of glaucoma. Therefore, the present inventors depended on the use of an agent for producing daltamate neuronal cell death as an index of an inhibitory effect on retinal ganglion cell death.

Since then, the present inventors have established a basic method based on the inhibitory effect on low-concentration daltamate neuronal cell death, which has been implicated in pathological conditions such as glaucoma. I've been experimenting. As a result, they have found that, in addition to the AMPA-KA receptor antagonist, dihydropyridine compounds such as ilvadipine also have an excellent cell death inhibitory action, and have reached the present invention. Hereinafter, the dihydric pyridine compound used in the present invention will be described.

In the dihydric pyridine compound (I), 2-nitrophenyl, 3-nitrophenyl, 4-nitrotropenyl and the like represented by R i are represented by R ² , R ³ and R ⁴ . Examples of the lower alkyl group include methyl, ethyl, propyl, isopropyl, butyl, tertiary butyl, pentyl, and hexyl.

Suitable salts of the dihydropyridine compound (I) are not particularly limited as long as they are pharmaceutically acceptable salts. For example, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfone Organic acid addition salts such as acid salts, formate salts and toluenesulfonate salts; inorganic acid addition salts such as hydrochlorides, hydrobromides, hydroiodides, sulfates, nitrates and phosphates Salts with acidic amino acids such as aspartate, glutamate, and the like;

The therapeutic agent for a disease associated with retinal ganglion cell death according to the present invention contains the above dihydropyridine compound (I), typically diluvadipine, or a salt thereof as an active ingredient, and contains an inorganic or organic carrier (medicine). Pharmaceutically acceptable carrier components), and is formulated into a solid, semi-solid, or liquid form into a dosage form such as an oral or parenteral dosage form.

As described above, the medicament of the present invention can be provided in any form of an oral preparation and a parenteral preparation, and an optimal dosage form can be selected according to the administration route, administration target, and the like. Examples of dosage forms suitable for oral administration include tablets, pills, powders, granules, soft and hard capsules, pellets, sublinguals, troches, and various liquids. Examples include injections, infusions, infusions, suspensions, oils, emulsions, suppositories, etc. Dosage forms suitable for administration include eye ointments, eye drops, sprays and the like.

When formulating the preparation using the active ingredient of the present invention, a conventional method may be used, and a surfactant, an excipient, a coloring agent, a flavor, a preservative, a stabilizer, a buffer, a suspending agent, etc. A toning agent and other commonly used carriers (which are not particularly limited as long as they are pharmaceutically acceptable) are appropriately used. In addition to the above, it is also possible to apply various new dosage forms known under the so-called DDS name.

The dose of the above dihydropyridine compound (I), typically, dilvazipine, is appropriately determined in consideration of the type of dosage form, administration method, age and weight of the patient, degree of symptoms, and the like. It is preferable to administer about 0.1 to: LOO mg, preferably 1 to 16 mg per day by oral administration. Effective doses are selected in the range of 0.001 to 1 mg / kg of patient weight; in the range of L mg, preferably in the range of 0.01 to 0.16 mg.

Hereinafter, the present invention will be described in detail based on examples. However, the following embodiments do not limit the present invention, and all changes and implementations without departing from the spirit of the preceding and following embodiments are included in the technical scope of the present invention.

[Experimental example]

Using cultured retinal ganglion cells, the inhibitory effect of ilvadipine on daltamate neuronal cell death was examined as follows. There were no safety issues.

1. Method

According to the method of Otori et al. (Invest. Ophthalmol. Vis. Sci, 39: 972-981, 1998), the retinal ganglion cells were transferred to the Long-Evans rat retina at 6 to 8 days of age by two-step immunopanning. Pure culture was performed. Specifically, using a serum-free medium containing neurotrophic factor and forskolin, simultaneously administer 25 M of glutamate (G lu) and nilvadipine (N iV,: ~ 100 nM) immediately after culture. After cultivating On day 3 of feeding, luciferin staining was performed, and the survival rate of retinal ganglion cells (RGC) was examined [mean soil standard deviation (%)].

2. Result

The results are shown in FIG.

In FIG. 1, “cont” is a control group cultured without administration of glutamic acid; “G1u” is a control group of Glu alone administered with only glutamic acid without administration of ilvadipine; “lu + Nivl 0 nM” is a group cultivated by simultaneous administration of glutamic acid and 100 nM of diluvadipine; “Glu + NivlOnM” is a group of glutamic acid and 100 ηM Group cultured with simultaneous administration of nilvadipine; “G1u + NiV1ηΜ” means a group cultured with simultaneous administration of glutamic acid and 1 nM of nilvadipine, respectively. In the figure, the “survival rate of RGC” is shown as a ratio when the control group is 100%.

From Fig. 1, the following can be considered.

First, the survival rate of retinal ganglion cells in the G1u alone administration group was 46.2 ± 5.9 (), which was about 1 Z2 lower than that in the control group. Nerve cell death was confirmed. This neuronal death was suppressed by administration of nilvadipine, and it was found that a concentration-dependent inhibitory effect was exhibited. In this experimental system, it was also confirmed that neuronal cell death due to glutamate was suppressed by administration of DNQX.Nilvadipine, as well as DNQX, a known daltamate neuronal cell death inhibitor, has a cell death inhibitory effect. It has become clear to have.

Example 1

Nilvadipine 100g

Hydroxypropyl methylcellulose 500g

Dissolve Nilvadipine in absolute ethanol (5 liters) and add hydroxypropyl methylcellulose to this solution to prepare a suspension. Next, the organic solvent is removed under reduced pressure to obtain a solid dispersion composition.

Example 2

Nilvadipine 100g Hydroxypropyl methylcellulose 500g

Sucrose 9.4kg

Add sucrose to a suspension of nilvadipine and hydroxypropylmethylcellulose in absolute ethanol (5 liters) and stir the resulting mixture. Next, the organic solvent is removed under reduced pressure to obtain a solid dispersion composition. This composition is made into fine granules by a conventional method.

Example 3

Nilvadipine 100g

Hydroxypropyl methylcellulose 500g

6.87Kg lactose

Low-substituted hydroxypropylcellulose 1.5kg

Magnesium stearate 30g

To a suspension of nilvadipine and hydroxypropylmethylcellulose in absolute ethanol (5 liters) was added lactose and lower substituted hydroxypropylcellulose, the resulting mixture was stirred, and the organic solvent was then depressurized. Removed below to obtain a solid dispersion composition. After granulating this composition by a conventional method, magnesium stearate is added to make a tablet by a conventional method. This tablet contains 2 mg of diluvadipine per tablet.

Example 4

Each tablet obtained in Example 3 was treated with hydroxypropyl methyl cellulose (5.1 mg), titanium dioxide (1.6 mg), polyethylene glycol 600 (0.8 mg), talc (0.4 mg), Film coating is carried out by a conventional method using a coating layer composed of yellow iron oxide (0.1 mg) to obtain a film coated tablet containing 2 mg of diluvadipine in one tablet. Industrial applicability

As described above, the drug of the present invention containing the dihydropyridine compound (I) represented by nilvadipine has an inhibitory effect on retinal ganglion cell death, and retinal degeneration associated with the ganglion cell death. Optic neuropathy (sight Neuritis, papillitis, optic retinitis, etamptor optic neuropathy, diabetic optic neuropathy, ischemic optic neuropathy, etc.) and glaucoma and other ophthalmic diseases can be expected to exert an effective therapeutic effect. It is also an excellent drug in terms of safety.

In the present invention, the inhibitory effect on retinal ganglion cell death is described, but the inhibitory effect of the dihydropyridine compound exerts an inhibitory effect not only on retinal ganglion cell death but also on other ganglion cell death. Various diseases associated with ganglion cell death, for example, psychiatric diseases such as retinal artery occlusion, depression, and epilepsy; central nervous system diseases such as aging-related neuropathy and dementia; Peripheral nerve region diseases such as vestibular nerve lesions such as dizziness and diabetic nerve lesions; various organ diseases caused by nerve cell lesions [eg, obstructive thrombotic vasculitis]

(Winiwarter-Buerger disease) and other circulatory diseases; skeletal muscular diseases such as amyotrophic lateral sclerosis, etc .; Demonstrates excellent clinical therapeutic effects in the treatment of visual abnormalities It is expected that.

Claims

Claims General formula (I)

(Wherein R ¹ represents nitrophenyl, and R ² , R ³ and R ⁴ each represent a lower alkyl group)

An agent for suppressing retinal ganglion cell death, comprising a dihydropyridine compound represented by the formula or a salt thereof as an active ingredient.

2. The retinal ganglion cell death inhibitor according to claim 1, wherein the dihydropyridine compound represented by the general formula (I) is ilvadipine.

3. General formula (I)

A therapeutic agent for a disease associated with retinal ganglion cell death, which comprises a dihydropyridine compound represented by the formula (1) or a salt thereof as an active ingredient.

4. The therapeutic agent according to claim 3, wherein the dihydropyridine compound represented by the general formula (I) is ilvadipine.

5. The therapeutic agent according to claim 3, wherein the disease is retinal degeneration.

6. The therapeutic agent according to claim 3, wherein the disease is optic neuropathy.

7. The therapeutic agent according to claim 6, wherein the disease is optic neuritis, papillitis, optic retinitis, etamptor optic neuropathy, diabetic optic neuropathy or ischemic optic neuropathy.