WO2004037246A1

WO2004037246A1 - Nerve fiber regeneration accelerator

Info

Publication number: WO2004037246A1
Application number: PCT/JP2003/013544
Authority: WO
Inventors: Masami Watanabe
Original assignee: Kowa Co., Ltd.
Priority date: 2002-10-24
Filing date: 2003-10-23
Publication date: 2004-05-06
Also published as: KR20050059274A; JP4675232B2; JPWO2004037246A1; AU2003275609A1

Abstract

A nerve fiber regeneration accelerator which contains 3,4-dihydro-8-(2-hydroxy-3-isopropylamino)propoxy-3-nitroxy-2H-1-benzopyran as an active ingredient.

Description

Description Nerve fiber regeneration promoter Technical field

The present invention relates to a nerve fiber regeneration promoting agent having an excellent nerve fiber regeneration promoting effect. Background art

Vertebrate nervous tissue is bisected into the central nervous system from the neural tube and the peripheral nervous system from the neural crest. The central nervous system consists of the brain, spinal cord and retina, and the peripheral nervous system includes the cranial nerves, spinal nerves, peripheral ganglia, and adrenal medulla. Thus, the two nervous systems are distinguished both embryologically and anatomically, but also differ in mammals in whether or not the severed nerve fibers (axons) regenerate.

Nerve fibers in the central nervous system of mature mammals do not regenerate when severed, but nerve fibers in the peripheral nervous system regenerate when conditions are met. This difference is not due to the differences in the properties of the cells of the central nervous system (neurons) and those of the peripheral nervous system (ganglion cells), but in the environment surrounding them, especially in the nerve fiber membrane. It is due to the difference in cells that form a certain myelin (oligodendrocytes in the center and Schwann cells in the periphery).

Diseases associated with damage to nerve fibers, particularly central nerve fibers, include many diseases caused by trauma and other causes in the brain, spinal cord, and optic nerve, and treatment by regeneration of nerve fibers is desired.

As for attempts to regenerate central nerve fibers, it is widely known that transplantation of peripheral nerves and detection of axonal regenerating cells by retrograde labeling are widely known (AJ AGUAYO: in Synaptic PI ascitics, CW). COTMAN ed., Guil ford, 1985,. 457-484). In the verification of functional recovery by axonal regeneration, the optic nerve of animals was cut and autologous transplantation of peripheral nerves was performed. By cross-linking with the center, the anti-miotic reflex (SJOWHITELEY et al .: Exp. Neurol., 1999, Vol. 154, .560) and avoidance behavior (H. SASAKI et al .: Exp. Neurol) , 1999, Vol. 159, p. 377). However, there are many issues that need to be overcome in the recovery of these functions. The synapse has not yet been formed. In particular, the number of regenerative fibers is extremely small, with only the optic nerve regenerating less than 4% of the total optic nerve fibers. In order for axons to regenerate, it is essential that nerve cells survive even if the axons are severed, but promotion of survival does not directly lead to promotion of axon regeneration (Goldberg〗 L , Barres BA: A. Rev. Neurosci., 2000, Vol. 23, p.579-612, Lu P et al .: J. Comp. Neurol., 2001, Vol. 436, p.456- 470, Watanabe M et al .: Invest. Ophthalmol. Vis. Sci., 2000, Vol. 41, S190)). In the same system as in the Examples below, we have obtained results that intraocular injection of neurotrophic factor and folskolin increased the number of viable cells, but did not promote axonal regeneration. Therefore, in order to promote axonal regeneration of the central nervous system by a drug, the drug is required to have an effect of increasing the number of surviving cells and also promoting axonal regeneration. However, the existence of a drug that has such an effect, promotes axonal regeneration of the central nervous system, and can be used for treatment has not been known until now.

On the other hand, 3,4-dihydro-8- (2-hydroxy-13-isopropylamino) propoxy-13-nitroxy-2H-1-benzopyran (generic name: niprazirol) has an excellent three-blocking action. It is known to be useful as a therapeutic agent for circulatory diseases such as hypertension and angina (Japanese Patent Publication No. 60-54317, Japanese Patent Publication No. 01 53245). There is no known effect of nipradilol on nerve fiber regeneration. Disclosure of the invention Accordingly, an object of the present invention is to provide a nerve fiber regeneration promoting agent having a nerve fiber regeneration promoting effect.

In view of the above-mentioned circumstances, the present inventor has conducted intensive studies, and surprisingly surprisingly found that Nipradilol, a known i3 blocker used for the treatment of hypertension and angina, has a function of nerve fiber regeneration. They have found that they have an accelerating action, and have completed the present invention. That is, the present invention provides a nerve fiber regeneration-promoting agent containing nipradilol as an active ingredient.

The present invention also provides use of nipradilol for producing an agent for promoting nerve fiber regeneration.

Furthermore, the present invention provides a method for promoting nerve fiber regeneration in a nervous system disease patient, which comprises administering an effective amount of nipradilol.

According to the present invention, a nerve fiber regeneration promoting agent having an excellent nerve fiber regeneration promoting effect can be provided. BEST MODE FOR CARRYING OUT THE INVENTION

Niprazilol used in the present invention can be produced by the method described in Japanese Patent Publication No. Sho 60-54317 as described above, and as described in Examples below, excellent nerve fiber regeneration promotion in a feline optic nerve transection model Since it has an effect, it is useful as a nerve fiber regeneration promoter.

The nerve fiber regeneration-promoting agent of the present invention contains Nipradi mouth as an active ingredient, and is an oral or parenteral agent such as an oral agent, an injection, a suppository, an ointment, a patch, and an eye drop. For example, a pharmaceutically acceptable carrier is blended as a composition suitable for the administration form, and the composition can be produced by a commonly used formulation method known to those skilled in the art.

When preparing oral solid dosage forms, for example, add excipients and, if necessary, binders, disintegrants, lubricants, coloring agents, Tablets, coated tablets, granules, powders, capsules and the like can be manufactured. So Such additives as may be those commonly used in the art. For example, excipients include lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose Water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, lipoxymethylcellulose, hydroxypropylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose , Shellac, calcium phosphate, polyvinylpyrrolidone, etc .; and as a disintegrant, dry starch, sodium alginate, powdered agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, lactose, etc. As a balm Ltd. talc, stearic acid salts, borax, polyethylene glycol, sucrose as a flavoring agent, orange peel, Kuen acid, can be exemplified tartaric acid.

When an oral liquid preparation is prepared, for example, an oral solution, a syrup, an elixir and the like can be produced by adding a flavoring agent, a buffering agent, a stabilizer, a deodorant and the like to nipradilol. In this case, the above-mentioned flavoring agents are preferred. Sodium citrate or the like is used as a buffer, and tragacanth, arabia gum, gelatin or the like is used as a stabilizer.

When preparing an injection, for example, add a pH adjuster, buffer, stabilizing agent, isotonic agent, local anesthetic, etc. to Nipradilol, and inject subcutaneously, intramuscularly and intravenously by the usual method Agents can be manufactured. In this case, examples of the pH adjuster and the buffer include sodium citrate, sodium acetate, sodium phosphate and the like. Examples of the stabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, thiolactic acid and the like. Local anesthetics include proforce hydrochloride, lidocaine hydrochloride and the like. Examples of the tonicity agent include sodium chloride, pudose and the like.

Formulations can be similarly formulated in other dosage forms according to known methods. The nerve fiber regeneration-promoting agent of the present invention obtained in this way can be used for any central nervous system damage or disease that causes or is accompanied by axonal damage, ie, the brain, spinal cord or optic nerve. It is effective in treating any part of the injury or disease including the sutra. In particular, trauma such as subinjury injury, anti-injury injury, penetrating trauma, trauma that occurs during neurosurgery or other procedures, stroke such as hemorrhagic stroke or ischemic stroke, optic neuropathy, glaucoma It is effective for treatment of optic nerve damage accompanying the above.

The dosage of the nerve fiber regeneration-promoting agent of the present invention varies depending on the patient's body weight, age, gender, symptoms, administration form, number of administrations, and the like. It is preferable to orally or parenterally administer を 100 mg, preferably 0.1〜10 mg in one or several divided doses.

Example

Hereinafter, the present invention will be described in detail with reference to Examples, but the present invention is not limited thereto.

Example 1

The nerve fiber regeneration promoting effect was evaluated according to the following procedure.

1. Anesthesia

Adult cats were anesthetized with a mixture of laughter (0.5 LZmin), halothane (1-: L. 5%), and oxygen (1 L Zmin), and fixed in a stereotaxic apparatus. Atosulfate pin 4 mg and penicillin 200,000 units were injected intraperitoneally and subcutaneously, respectively.

2. Injection of Nipradilol

A small hole was made in the sclera behind the serrated margin of the left eyeball with a 25 G needle. The volume of the vitreous average 2. 7 ml, so that intravitreal concentration of the two Purajiroru becomes ^{1 X 10_ 7 mo 1 / L} , dissolved in Ringer's solution to adjust the concentration. 10 L of an aqueous solution was injected into the vitreous body from the stoma using a 10 / xL, Milton syringe. As a control, a group without injecting nipradilol was prepared.

3. Optic nerve transection and peripheral nerve transplantation

The common peroneal nerve (40-5 Omm long) was removed from the thigh of the same individual. 4. For the convenience of marking, use 10-0 composite yarn at 1 Omm and 2 Omm from the center end. Marked.

Thirty to sixty minutes after injection of nipradilol, the optic nerve was completely cut immediately after the eyeball and the stump was sutured with the common peroneal nerve. The other end was placed intramuscularly.

4. Labeling of regenerative cells

Twenty-four days after transplantation of the peripheral nerve (4 days before the fourth week) or 38 days (4 days before the sixth week), the transplanted nerve was excised under anesthesia. Using the 10-0 suture as a clue, cut the graft at 2 O mm from the joint, place the red fluorescent dye dexstran-rhodamine powder on the stump, and close with the 10-0 suture. Was. Two days later (two days before the experimental treatment), the transplanted nerve was cut at a point 10 mm from the junction, and a yellow fluorescent dye dexs tran-fluorescein was placed on the stump, and 10-0-0 鏠Closed with twine.

5. Total cell count estimation

Four or six weeks after transplantation of the peripheral nerve, the cat was deeply anesthetized with Nembu, and the left eyeball was enucleated. The retina was excised in the Ames culture and fixed with 3% balformaldehyde-phosphate buffer.

Progressive specimens were prepared by enclosing the retina in PennaFluor, and the total number of cells and the double labeling rate were determined using a fluorescence microscope. Fluorescein-labeled cells (cells with axons extending from 10 to 20 mm, 10R) within 480 m square and cells labeled with both florescein and rhodamine (axons Cells extending 20 mm or longer, 20 R) were counted at 1 mm intervals to estimate the total cell number. 20 RZ 10 R was defined as the double labeling rate and used as a measure of the axonal outgrowth rate. Table 1 shows the results of the total cell number and the results of the double labeling ratio.

table 1

As shown in Table 1, the total number of cells in the control was 4 weeks after transplantation of the peripheral nerve. W 200 and 2139 ± 795 units, respectively after 6 weeks, against to a 313 amino ± 3678, two Purajiroru (final concentration: 10- ⁷ mo 1 / L) by intravitreal injection, total cell number is 4 weeks The number increased to an average of 8963 later, and 11278 ± 5568 after 6 weeks. In addition, 2OR / 1OR indicates the percentage of cells regenerating axons of 20 mm or more in cells that regenerated axons of 1 Omm or more. after contrast is the respectively 17 ± 4% and 65 ± 10%, two Purajiroru (final concentration: 10- ⁷ mo 1ZL) in retinal injected intravitreally with each 38% after after 4 weeks and 6 weeks and It increased to 77 ± 14%. From the results shown in Table 1, promotion of axonal regeneration by nipradilol was confirmed.

Claims

The scope of the claims

1. A nerve fiber regeneration promoter containing 3,4-dihydro_8- (2-hydroxy-13-isopropylamino) propoxy-3-nitroxy-2H-1-benzopyran as an active ingredient.

2. The nerve fiber regeneration promoter according to claim 1, wherein the nerve fiber is a central nerve fiber.

3. The nerve fiber regeneration-promoting agent according to claim 2, wherein the central nerve fiber is an optic nerve fiber.

4. Use of 3,4-dihydro-8- (2-hydroxy-3-isopropylamino) -butoxy-3-nitroxy_2-H_1-benzopyran for producing nerve fiber regeneration promoter.

5. The use according to claim 4, wherein the nerve fiber is a central nerve fiber.

6. The use according to claim 5, wherein the central nerve fiber is an optic nerve fiber.

7. The promotion of nerve fiber regeneration in patients with nervous system diseases characterized by administering an effective amount of 3,4-dihydro-8- (2-hydroxy_3-isopropylamino) proboxy-3-ditroxy-2H-1-benzopyran Method.

8. The method according to claim 7, wherein the nerve fiber is a central nerve fiber.

9. The method according to claim 8, wherein the central nerve fiber is an optic nerve fiber.