MXPA00006353A - Remedy for neurodegenerative diseases___________________________ - Google Patents

Abstract

A remedy for neurodegenerative diseases containing zonisamide or an alkali metal salt thereof as the active ingredient and usable for preventing and remedying mammalian (including human) neurodegenerative diseases such as Parkinson's disease, Huntington's disease, choreic syndrome and dystonic syndrome.

Description

MEDICINE FOR NEURODEGENERATIVE DISEASES TECHNICAL FIELD The present invention relates to a medicament for neurodegenerative diseases comprising zonisamide or an alkali metal salt thereof as an active ingredient.

ANTECEDENTS OF THE TECHNIQUE Zonisamide [chemical name: 3-sulfamoylmethyl-l, 2-benzisoxazole or 1,2-benzisoxazole-3-methanesulfonamide; see, for example, Merck Index, 12th Ed., 10323 (1996)] has been used as an antiepileptic agent in the treatment or prevention of various attacks in Japan and South Korea, etc. JP-B-60-3311, JP-B-61-59288 and US 4,172,896 describe processes for preparing zonisamide and the utility thereof as an anti-epileptic agent. In addition, JP-B-7-84384 and USP 5,128.35 also describe the utility of zonisamide as a medicament for ischemic brain damage. As more and more aging societies develop, the number of patients suffering from neurodegenerative diseases such as Parkinson's disease increases. Parkinson's disease is a disease REF.121051 progressive and tragic by which the coordinated movement of the patient is altered, the coordinated movement of the patient is altered, and the patient's movement becomes slow with the passage of time and, finally, rigidity or tremor develops in the arms and legs . It has been known that the disease is caused by suppression of dopamine in the fetus due to rhexis and loss of dopamine-producing neurons in the nigro-estpatal region. Incidentally, an animal lacking a stepatal dopamma, which is prepared by administering l-met? L-4-phen? 1-2, 3, 6-tetrah? Drop? Pdma. { hereinafter referred to as "MPTP") to a C57 black mouse, it has been widely used as an animal model for Parkinson's disease. Life Sci. , 54, 245 (1994) describes the antiepileptic agents: lamotrigine [chemical name: 3,5-d? Ammo-6- (2, 3-d? Chlorophen? L) -1, 2, 4-tpaz? Na ] and phenytoin show an inhibitory effect on the suppression of doparrane induced by MPTP, while carbamazepma has no such effect. However, in the literature experiments, MPTP hydrochloride has been injected subcutaneously at a dose of 15 mg / kg only once to C57 black mice, and therefore such mice are not necessarily suitable as an animal model for Parkmson's disease. In the current Parkmson's disease medication, agents that replenish dopamma are used (for example, levodopa alone, or a combination product, or combination therapy of levodopa and carbidopa), dopamine agonists (for example, bromocriptma or tergupde), dopamine releasing agents (for example amantadma), anticolmergic agents (for example bipepden or trihexife) idyl), inhibitors of monoammo oxidase type B (MAO-B) (for example selegiline), etc. However, these agents are not necessarily satisfactory from the point of view of efficacy and side effects, and it has been expected to develop a novel effective medicament. The present inventors have found that the zonisamide and an alkali metal salt thereof show an extremely potent inhibitory effect in the dopammérgica neurodegeneration induced by MPTP and have carried out the present invention.

DESCRIPTION OF THE INVENTION An object of the present invention is to provide a medicament for the treatment of neurodegenerative diseases comprising zonisamide or an alkali metal salt thereof as an active ingredient. Another objective of the present invention is to provide in use of zonisamide as a metal salt alkaline thereof for the manufacture of a medicament for neurodegenerative diseases. A further objective of the present invention is to provide a method for the prevention and / or treatment of neurodegenerative diseases in a mammal (including the human), which comprises administering an effective amount of zonisamide or an alkali metal salt thereof to the mammal. in need of such prevention and / or treatment BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph showing the content of various monoamine neurotransmitters and metabolites thereof in the excised stool 24 hours after the last MPTP administration of male C57 black mice, to which MPTP hydrochloride has been administered intraperitoneally. once a day at a dose of 30 mg / kg for 8 days, repetitively. Figure 2 is a graph showing the inhibitory effect of zonisamide and carbamazepma on MPTP-induced dopaminergic neurodegeneration in C57 black male mice 14 days after the last MPTP administration.

BEST MODE FOR CARRYING OUT THE INVENTION The alkali metal salts of zonisamide are sodium salt, potassium salt and lithium salt. Zonisamide can be prepared, for example, by the methods described in JP-B-60-33114, JP-B-61-59288 and USP 4,172,896. The results of the pharmacological experiments of zonisamide and of known antiepileptic agents are shown below, and therefore the usefulness of zonisamide as a medicament for neurodegenerative diseases will be explained. The following are the antiepileptic agents used as reference drugs in pharmacological experiments. Carbamazepine (see, for example, Merck Index, 12th Ed., 1826 (1996)); Phenytoin (see, for example, Merck Index, 12th, Ed., 7475 (1996)); Lamotrigine (see, for example, Merck Index, 12th Ed., 5367 (1996)); Phenobarbital (see, for example, Merck Index, 12th Ed., 7386 (1996)); Ethosuximide (see, for example, Merck Index, I2th Ed., 3794 (1996)); Sodium valprophate (see, for example, Merck Index, 12th Ed., 10049 (1996)); Diazepam (see, for example, Merck Index, 12th Ed., 3042 (1996)); and Acetazolamide (see, for example, Merck Index, 12th Ed., 50 (1996)). In each experiment, the content of the monoamine neurotransmitters and the metabolites thereof in the striatum are measured by the high-performance liquid chromatography (HPLC) method. HPLC is carried out using a system consisting of a pump (L-6000, manufactured by Hitachi, Ltd., Japan), an autoinjector, a column (Cosmosil 5C18-A, manufactured by Nacalai Tesque, Inc., Japan; 4.60 x 250 mm), and an electrochemical detector (ECD-100, manufactured by Eicom Corporation, Japan) under the following conditions.

Mobile phase: 0.05M citric acid containing 0.075 M sodium perchlorate buffer: acetonitrile (92.5: 7.5, pH 4. 3) . The buffer contains sodium octasulfonate 0. 022% and disodium salt of EDTA.2 0.0015%.

Flow rate: 0.9 ml / min.

Conditions for Detection: Working Electrode: Graphite Electrode Reference Electrode: Silver / Silver Chloride Applied voltage: 750 mV In addition, the statistical analysis is carried out by Dunnett's multiple comparison test.

Experiment 1 Preparation of the animal model for Parkinson's disease: Male C57 black mice (11 weeks old) are used in the experiment. MPTP hydrochloride is dissolved in physiological saline at a concentration of 3 mg / ml, and the resulting solution is injected intraperitoneally into the mice in a volume of 0.1 ml per 10 g of body weight, i.e. at a dose of 30 mg / kg, once a day for 8 days, repetitively. After 24 hours of the last administration of MPTP, the head of the mice is subjected to microwave irradiation, and the striatum is removed from it. The striatum removed is homogenized with 50 times the volume of 0.1 N formic acid: acetone (15:85) containing an internal standard substance (5-hydroxytryptopol, 20 ng / ml), and the mixture is centrifuged under cooling. A fixed amount of the supernatant is collected and evaporated to dryness under a stream of nitrogen gas. Subsequently, the resulting substance is dissolved in 0.01 N acetic acid and centrifuged, and then the content of various monoamine neurotransmitters and metabolites thereof in the supernatant are measured by HPLC which is equipped with an electrochemical detector. Changes in the content of various neurotransmitters and metabolites thereof 24 hours after the last MPTP administration are shown. in figure 1 The number of animals in each group is 8, and the lines perpendicular to the figures represent standard errors, and the mark ** means that there is a significant difference with respect to the value of the control group (the group treated with a physiological saline solution) with p 0 01 As it is evident from the figure 1, MPTP significantly reduces the content of dopamma (DA) and metabolites thereof, homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC), but never affects the content of noradrenaline (NA), serotomna (5HT) and metabolites thereof, 3-methox? -4-h? drox? phen? let? lengl? col (MOPEG) and 5-hydroxnindolacetic acid (5HIAA) and therefore it is confirmed that MPTP shows selectivity for dopammergic neurons. that the reduction in the dopamine content and metabolites thereof in the MPTP-induced estpate is observed even 14 days after the last administration of MPTP, as it is clear from figure 2 that shows the results of experiment 2, this reduction in the content of it is irreversible, and it is suggested that the mice that have been suppressed dopamma are extremely suitable as an animal model of Parkmson's disease Experiment 2 Inhibitory effect of dopammérgica neurodegeneration induced by MPTP: It is administered mtraperitoneally hydrochloride MPTP to C57 black male mice (11 weeks old) at a dose of 30 mg / kg, once a day for 8 days, repetitively in the same manner as in experiment 1. Test drugs, ie, anti-epileptic agents , are suspended in an aqueous solution of tragacanth at 0.5%, and the resulting solution is orally administered to the mice in a volume of 0.1 ml per 10 g of body weight, 30 minutes before the administration of MPTP for 8 days. hours and 14 days later, after the last administration of MPTP in the zonisamide and carbamazepma experiment, the head of the mice is subjected to microwave irradiation, and the estpate is removed from it. The content of dopamma and its metabolites in the estpate is measured in the same way as in experiment 1. In the experiments of various antiepileptic agents, zonisamide (ZNS), carbamazepma (CBZ) and the group consisting of phenitoma (PHT), lamotpgma (LTG), phenobarbital (PB), ethosuximide (ESM), sodium valproate (VPA), diazepam (JZP) and acetazolammda (AZA) They are tested on different days. The dosages of each administration of various antiepileptic agents are shown below. In addition, the values in parentheses after the dosages mean the ratio with respect to the dose that the anticonvulsant activity shows in mice. Since the doses show anticonvulsant activity, the values described in Epilepsy 27, 483 (1986) for lamotrigine and diazepam, and the values described in Arzneim, Forsch, (Drug Res.), 3_0, 477 (1980) for other antiepileptic agents, are the ones that were used respectively. Zonisamide (ZNS) 10, 30, 100 mg / kg (approximately 0 5 times, approximately 1.5 times, approximately 5.1 times, respectively) Carbamazepma (CBZ) • 20 and 60 mg / kg (approximately 1.5 times, and approximately 4.5 times, respectively ) Phenitoma (PHT). 30 mg / kg (approximately 3.8 times) Lamotrigma (LTG) 30 mg / kg (approximately 11.5 times) Phenobarbital (PB): 40 mg / kg (approximately 3.4 times) Ethosuccimide (ESM). 600 mg / kg (approximately 2.2 times) Sodium valproate (VPA). 800 mg / kg (approximately 2.5 times) Diazepam (DZP) 5 mg / kg (approximately 4.2 times) and Acetazolamide (AZA). 100 mg / kg (approximately 3.8 times). Table 1 shows the inhibitory effect of the various antiepileptic agents on dopammérgica neurodegeneration 24 hours after the last administration of MPTP, from the point of view of the content of dopamma and the metabolite thereof, acid 3, 4-d? H? drox? phenylacetic (DOPAC), in the estpato, and at an inhibitory speed. The inhibitory rate is calculated from values in a group of normal animals, a group treated with vehicle and the group treated with the test drug. The values in the table represent an average value ± a standard error, and the marks * and ** mean that there is a significant difference with respect to the value of the group treated with vehicle, with p < 0.05 and p < 0.01, respectively. In addition, Figure 2 shows the inhibitory effect of zonisamide and carbamazepma on dopammérgica neurodegeneration 14 days after the last administration of MPTP (5 to 6 animals in each group). The perpendicular lines in the figures represent standard error, and the marks * and ** mean that there is a significant difference with respect to the value of the group treated with vehicle, with p < 0.05 and p < 0.01, respectively Table 1 MPTP-induced inhibitory effect of dopaminergic neurodegeneration Test drug Dosage Number of DA Speed DOPAC Rate (mg / kg, po) animals (ng / g of tissue) Inhibitory (%) (ng / kg of tissue) Inhibitory (%) Group with normal animals 8598 + 560 * Group treated with vehicle 2366+ 141 9 3797 + 201 * 22.3 221 + 11 * 29.8 30 16 7373 + 685 * '80.3 370 + 27 ** 82.1 100 8 8434 + 336 ** 97.2 447 + 24 ** 109.1 Group with normal animals 6558 ± 743 * Group treated with vehicle 1932 + 355 3087 + 453 25.0 251 + 36 40 6 2894 + 750 20.8 202 + 52 23 0 Group with normal animals 7485 + 705 * Group treated with vehicle 4073 + 454 * 37.4 257 + 30 * 30 2 LTG 4844 + 676 ** 51.6 273 + 31 ** 34 1 3310 + 544 23.5 200 + 32 16 3 1008 + 182 2984 + 468 175 187 + 29 3356 + 451 1803 + 428 As is evident from Table 1 and Figure 2, the zonisamide (ZNS) of the present invention shows a dose-dependent inhibitory effect on the reduction induced by MPTP in the dopamine (DA) content and the metabolite thereof, acid 3, 4-d? h? drox? phen? lac? co? co (DOPAC), in striatum both in the 24-hour group and in the 14-day group, from a dose of 10 mg / kg which it is approximately 0.05 times the dose that shows anticonvulsant activity. Especially, zonisam shows an extremely high inhibitory effect in the reduction in the content of both dopamma and DOPAC at a dose of 30 mg / kg (a dose which is approximately 1.5 times the dose showing anticonvulsant activity), and sample almost 100% inhibitory rate at a dose of 100 mg / kg (a dose which is approximately 5.1 times the dose showing anticonvulsant activity). On the other hand, as is evident from Table 1 and Figure 2, carbamazepma (CBZ) does not show any significant inhibitory effect on MPTP-induced reduction in the content of dopamma and DOPAC in the estpate both in the 24 hours as in the group of 14 days. In addition, among the other antiepileptic agents, others other than lamotrigma (LTG) and phenytoin (PHT) may not inhibit the reduction in dopamine and DOPAC content in the stomat even at a dose which is twice or greater in comparison at the dose showing the anticonvulsant activity. Although the phenitoma significantly inhibits the reduction in the content of dopamma and DOPAC at a dose of 30 mg / kg (a dose which is approximately 3.8 times the dose that shows anticonvulsant activity), the inhibitory speeds of these are only 37.4% and 30.2%, respectively. Lamotine inhibits the reduction in the content of dopamma and DOPAC more strongly than the phenytoin at a dose of 30 mg / kg (a dose which is approximately 11.5 times the dose showing anticonvulsant activity), but the inhibitory rates thereof they are only 51.6% and 34.1%, respectively, which are much weaker than the inhibitory rates of 80.3% and 82.1% zonisamide, at a dose of 30 mg / kg (a dose which is approximately 1.5 times the dose that shows the anticonvulsant activity).

As is clear from the results of the above experiments, zonisamide and alkali metal salts thereof show extremely potent inhibitory effects on MPTP-induced dopaminergic neurodegeneration at a dose showing anticonvulsant activity with low toxicity, and therefore , can be used as a medicine for neurodegenerative diseases in prevention and treatment of various neurodegenerative diseases such as Parkinson's primary or secondary disease, Huntmgton's disease, choreic syndrome and dystonic syndrome in mammals (including humans). The medicament of the present invention can be administered through any route, oral, parenteral and intrarectal. The dosage of zonisamide or the alkali metal salts thereof can vary based on the route of administration, the classes of diseases being treated, the severity of the symptoms, the age of the patients, etc., but is usually in the range of 1 to 50 mg / kg / day, preferably in the range of 2 to 20 mg / kg / day, which can be administered once or several times. The zonisamide and the alkali metal salts thereof can be used as a medicament for neurodegenerative diseases alone or in the form of a pharmaceutical composition, which is prepared by mixing it with a pharmaceutically acceptable carrier or diluent. The pharmaceutical composition can be in dosage forms such as tablets, capsules, granules, powders, syrups, injection preparations and suppositories, and can be prepared by conventional methods. In addition, zo isamide tablets and powders, which are commercially available as an anti-epileptic agent, can be used as a medicament for neurodegenerative diseases of the present invention. The pharmaceutically acceptable carrier or diluent can be any of the conventional ones, which are commonly used in the pharmaceutical field and do not react with zonisamide or the alkali metal salts thereof. Suitable examples of pharmaceutically acceptable carriers or diluents for the preparation of tablets, capsules, granules and powders include excipients (for example lactose, corn starch, sucrose, mannitol, calcium sulfate or crystalline cellulose), disintegrants (for example sodium carmellose). , modified starch or calcium carmellose), binders (for example methylcellulose, gelatin, acacia, ethylcellulose, hydroxypropylcellulose or polyvinylpyrrolidone) and lubricants (for example light anhydrous silicic acid, magnesium stearate, talc or hydrogenated oil). The tablets can be coated in any conventional manner by the use of conventional coating agents such as carnauba wax, hydroxypropylmethylcellulose, macrogol, hydroxypropyl methyl phthalate, cellulose acetate phthalate, sucrose, titanium oxide, sorbitan fatty acid ester and phosphate calcium. Suitable examples of the pharmaceutically acceptable carrier or diluent for the preparation of syrups include sweetening agents (eg sucrose, glucose or fructose), suspension improving agents (eg, acacia, tragacanth, carmellose sodium, methylcellulose, sodium alginate, crystalline cellulose or Veegum), and dispersing agents (for example sorbitan fatty acid ester, sodium lauplsulfate or polysorbate 80). In the preparation of syrups, you can optionally adding a flavoring agent, a perfume or a preservative. In addition, such syrups may be in the form of a dry syrup, which is dissolved or suspended when used. Suitable examples of the suppository base include cocoa butter, saturated fatty acid ester and glycerin, glycerogelatine and macrogol. In the preparation of suppositories, optionally a surfactant or a preservative can be added. The preparations for injection are usually prepared by dissolving an alkali metal salt of zonisamide in distilled water for injection, and to this a solubilizer, a buffering agent, a pH adjusting agent, an isotonic agent, an agent for reduce pain or a conservative. These pharmaceutical compositions may contain zonisamide or an alkali metal salt thereof as an active ingredient in an amount of at least 0.5% (% by weight, below as the same), preferably 10-70%, based on the total weight of the composition. Optionally, these pharmaceutical compositions may contain other therapeutically effective compounds as mentioned below.

The medicaments for neurodegenerative diseases of the present invention can be administered together with other medicaments such as agents that replenish dopamine (for example levodopa alone, or a combination product or combination therapy of levodopa and carbidopa), dopamine agonists (for example bromocriptine or terguride), dopamine releasing agents (for example amantadine), anticholinergic agents (for example biperiden or triexyphenidyl) and inhibitors of monoamine oxidase type B (MAO-B) (for example selegiline). The pharmaceutical compositions of the medicaments for neurodegenerative diseases according to the present invention are exemplified below.

Preparation 1.- Tablets: Zonisamide 100 g Lactose 35 g Corn starch 17 g Crystalline cellulose 40 g Hydroxypropyl cellulose 6 g Light anhydrous silicic acid 1 g Magnesium stearate 1 g Total 200 g Among the above components, zonisamide, lactose, corn starch and crystalline cellulose are combined, to the combination is added hydroxypropylcellulose that is dissolves in water, and the mixture is kneaded, dried and granulated. Magnesium stearate and light anhydrous silicic acid are added to these granules, and the mixture is compressed to provide 1000 tablet cores weighing 200 mg each. Subsequently the tablet cores are coated to form film-coated tablets, by a conventional method, using hydroxypropylmethylcellulose, macrogol, titanium oxide, talc and light anhydrous silicic acid.

Preparation 2: Powders 20% Zonisamide 200 g Lactose 719 g Hydroxypropyl cellulose 20 g Light anhydrous silicic acid 1 Total 940 g By using a high shear granulator, the above components are combined, sprayed with an ethanolic solution (200 g, containing 40 g of ethylcellulose and 20 g of hydroxypropylcellulose for granulation), and granules. They are regulated in size to provide 20% powders.

INDUSTRIAL APPLICABILITY As explained above, zonisamide and the alkali metal salts thereof show a potent inhibitory effect on dopaminergic neurodegeneration at a dose showing anticonvulsant activity and therefore, are useful as a medicament for neurodegenerative diseases in the prevention and treatment of various neurodegenerative diseases such as Parkinson's disease, Hungtinton's disease, choreic syndrome and dystonic syndrome in mammals (including humans). It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers.

Claims (6)

1. Having described the invention as above, the content of the following claims is claimed as property 1 A medicament for neurodegenerative diseases, characterized in that it comprises zonisamide or an alkali metal salt thereof as an active ingredient
2. The medicament according to claim 1, characterized in that the active ingredient is zonisamide
3. The medicament according to claim 1 or 2, characterized in that the neurodegenerative disease is Parkmson's disease
4. 4 The use of zonisamide or an alkali metal salt thereof for the preparation of a medicament for neurodegenerative diseases
5. 5 The use of zonisamide for the preparation of a drug for neurodegenerative diseases
6. 6. The use according to claim 4 or 5, wherein the neurodegenerative disease is Parkinson's disease.