MXPA00003956A - Agent enhancing cerebral acetylcholine release - Google Patents

Abstract

A pharmaceutical composition comprising N-anisoyl-g-aminobutyric acid (also known as N-anisoyl-GABA or 4-[(4-methoxybenzoyl) amino]-butanoic acid ) or para-anisic acid (4-methoxybenzoic acid)enhances acetylcholine release in the brain and as a consequence, it may be used in the treatment of various neuropsichiatric disorders associated with a deficit of acetylcholine. Such disorders include dementia, circadian rhythm disorders, attention deficit disorder, sleep disorders and problematic behaviours such as delirium and nocturnal wandering. A lack of acetylcholine also occurs in patients with neuronal degradation resulting from conditions such as Alzheimer's, Parkinson's or progressive supranuclear palsy. The composition is preferably formulated in tablet or capsule form as a unit dosage of 1-300 mg/kg per adult per day. N-Anixoyl-g-aminobutyric acid and para-anisic acid are the major in vivo metabolites of aniracetam (1-para-anisoyl-2-pyrrolidinone).

Description

AGENT TO IMPROVE THE LIBERATION OF CEREBRAL ACETILCOLINE The present invention relates to the use of N-anisoyl-α-aminobutyric acid (N-anisoyl-GABA) or p-anisic acid, in the form of a pharmaceutical composition that enhances the release of acetylcholine (ACh) in the brain . In particular, the invention relates to a pharmaceutical composition for circadian rhythm disorders, sleep disorders, attention deficit disorders and problem behaviors. It has been thoroughly demonstrated that ACh is an important classic neurotransmitter. The main cholinergic pathway exists in mesopontin reticulum nucleus-thalamus, septum hippoca pus and basal nucleus neocortex of the forebrain in mammalian brains [Neuroscience, 1_0, 1185-1201 (1983)]. These 0 cholinergic routes have been thought to play a functionally pivotal role in the induction and maintenance of sleep with rapid eye movement (REMS), and in the regulation of vigilance and attention, learning and memory, and motivation 5 [Journal of Neuroscience, 1_0, 2541-2559 (1990); Brain REF..119438 Research Review, 1_9, 298-318 (1994)]. On the other hand, central cholinergic neuronal activity decreased in patients with neuronal degeneration (eg, Alzheimer's disease, Parkinson's disease and progressive supranuclear palsy) [Lancet, 2, n ° 8000, 1403 (1976); Journal of Neurological Neurosurgical Psychiatry, 5_1, 540-543 (1988)] and with cerebrovascular diseases [Dementia, 5, 163-167 (1994); Journal of Neural Transmission, 103, 1211-1220 (1996)]. It has been suggested that cholinergic deficit and / or dysfunction are associated with various neuropsychiatric symptoms, such as dementia, sleep disorder, low vigilance, attention deficit and problematic behaviors including delirium and nocturnal fantasies. L-p-anisoyl-2-pyrrolidinone (aniracetam, European patents n ° 5143 and 44068), an enhancer of brain function, has been used for the treatment of emotional disorders (anxiety / irritation and depressive moods) that appear as sequelae after a cerebral infarction [Igaku No Ayumi, 156, n ° 2, 143-167 (1991); Geriatric Medicine, 3_6, 1513-1520 (1998)]. Although the drug has been reported to activate central cholinergic systems, it remains to be clarified which substances, including their metabolites, are responsible for cholinergic activation when orally administered lp-anisoyl-2-pyrrolidinone [Drug Inves tigation , 5_, 1-108 (1993)]. In addition, the biological activities of N-anisoyl-GABA and p-anisic acid are not clear, nor are the main metabolites of l-p-anisoyl-2-pyrrolidinone. Meanwhile, treatment for the diseases described above has begun by raising central ACh levels with ACh esterase inhibitors. Because ACh esterase is widely distributed not only in the brain but also in peripheral tissues, the peripheral side effects caused by the poor penetration of ACh esterase inhibitors through the blood-brain barrier can cause serious problems for these enzyme inhibitors like medicines. Therefore, the problem to be solved by the present invention has been to provide promising compounds, which are capable of safely increasing central ACh levels. It has now been discovered that surprisingly N-anisoyl-GABA and p-anisic acid, potentiate each of them, the release of cerebral ACh in the brain. Thus, the present invention relates to the use of N-anisoyl-GABA or p-anisic acid, preferably in the form of a pharmaceutical composition, for the enhancement of the release of ACh in the brain of mammals, preferably humans. These compounds and pharmaceutical compositions containing N-anisoy1-GASA or p-anisic acid are of particular utility for the treatment of circadian rhythm disorders, sleep disorders, attention deficit disorders and problem behaviors. The invention relates to pharmaceutical compositions for the enhancement of the release of cerebral acetylcholine, which contain N-anisoyl-GABA or p-anisic acid as an effective ingredient for the treatment of the above-mentioned disorders. The invention also relates to the methods of using N-anisoyl-GABA or p-anisic acid for the enhancement of Ach release in the brain, in particular for the treatment of circadian rhythm disorders, sleep disorders, disorders of attention deficit and problematic behaviors. The present invention is explained in detail below. The description and examples presented below will help to understand the invention, but can not be taken as limiting the invention in any way. The results obtained in the examples are documented by the figures that accompany them. A brief description of them follows: Figure 1: Effects of aniracetam (A), 2-pyrrolidone, p-anisic acid and N-anisole-GABA (B) on the release of ACh in the rat nucleus of the rat thalamus spontaneously hypertensive patients prone to stroke (SHRSP) moving freely. The compounds were perfused for 20 minutes (strong trace). The data represent means of ± S.E.M. The baseline ACh release was 0.35 ± 0.03 pmol / 20 minutes (n = 24). * P <; 0.05 compared to the control of the vehicle. Figure 2: Effects of amracetam (A), 2-pyrrolidone, p-anisic acid and N-amsoyl-GABA (B) on the release of ACh in the dorsal hippocampus of SHRSP rats with freedom of movement. The compounds were perfused for 20 minutes (thick line). The data represent means of ± S.E.M. The baseline ACh release was 0.63 ± 0.04 pmol / 20 minutes (n = 15). * P < 0.05, ** P < 0.01 compared to the aniracetam. Figure 3: Effects of aniracetam (A), 2-pyrrolidinone, p-anisic acid (B) and N-anisoyl-GABA (C) on the release of ACh in the prefrontal cortex of SHRSP rats with freedom of movement. The compounds were perfused for 20 minutes (thick line). The data represent means of ± S.E.M. The baseline ACh release was 0.58 ± 0.03 pmol / 20 minutes (n = 24). * P < 0.05, ** P < 0.01 compared to the aniraceta. Figure 4: The rhythm of circadian motor activity and the anticipatory activity associated with feeding time in young and old rats. After a 24-hour fast (day 0), food was restricted to only 1 hour from 13.30 for 6 consecutive days (days 1 to 6). The food was retained again on day 7. The data show means of S.E.M. of the motor activity measured every hour, obtained from 7 rats for each group.

The dotted column indicates the period of the meal. ? = day -1; 0: = day 6; •: = day 7 Figure 5: Effects of aniracetam on the circadian motor and the rhythms of anticipatory activity associated with feeding time in old rats. Aniracetam was administered orally to the rats immediately after finishing the feeding once a day for 7 consecutive days (days 1 to 7). The data show means ± S.E.M. obtained from 5-6 rats per group. The column of points indicates the feeding period. 0: day 6; •: = day 7. Figure 6. Day and night changes of REMS, non-REMS (NREMS) and brain temperature in SHRSP and istar Kyoto rats of similar age (WKY). The data show means ± S.E.M. of each variable measured every hour. Figure 7: Effects of repeated administration of aniracetam on REMS, NREMS and brain temperature in SHRSP. The aniracetam was administered orally to the animals twice a day (9:00 and 20:00) for 5 consecutive days and the data are shown after dosages 9a and 10a.

Both N-anisoyl-GABA and p-anisic acid are chemically known compounds. N-anisoyl-GABA can be synthesized by methods described in Spanish patent publication No. 84-538772. P-anisic acid can be obtained by methods described in the Journal of the American Chemical Society (78), 907-909 (1956) and also purchased from Sigma Chem. Co. (St. Louis, USA), Lancaster Synthesis Ltd. (Lancashire, UK), Wako Puré Chem. Id. Ltd. (Osaka, Japan), etc. The N-anisoyl-GABA and the p-anisic acid can each be employed in the form of a pharmaceutically acceptable preparation. This preparation can be obtained in tablets, coated tablets, chocolates, hard gelatin capsules, soft gelatine capsules, as well as in solution, emulsion or suspension. The resulting formulation can be administered orally. In addition, this preparation can be obtained in suppositories for intrarectal administration, or in the form of an injectable to be administered parenterally. When producing peroral solid preparations, such as tablets, coated tablets, candies or hard gelatine capsules, N-anisoyl-GABA or p-anisic acid can each be formulated together with chemically inert inorganic or organic fillers, such as lactose, corn or corn starch and its derivatives, talc, stearic acid and its bases or salts, etc. When soft or hard gelatin capsule products are prepared, they can be appropriately used as fillers, for example, vegetable oils, waxes, fats, oils, gels, semisolid or liquid polyols, etc. When liquid products and syrups are prepared, fillers such as water, polyols, sucrose, invert sugar, glucose, etc. can be appropriately used. When injectable products are prepared, they can be appropriately used as fillers, water, alcohols, polyols, glycerols, vegetable oils, etc. When suppositories are prepared, they can be appropriately used as fillers, for example, vegetable oils, waxes, oils, gels or liquid polyols, etc. In addition, these preparations can be used in combination with antiseptics, solvents, stabilizers, humectants, emulsifiers, sweeteners, bases for modifying the osmotic pressure, buffers, epiboly and -antioxidants, and in addition, a therapeutically valuable compound. The route of administration of the previous preparation is not intended to be limited thereto, but it can be varied appropriately depending on the forms of preparation or age, sex, symptoms of the patients, etc. The route of administration, dosage and number of administrations can be varied appropriately depending on the age, weight and symptoms of the patients. In the case of oral administration, the dosage is usually 1 to 300 mg / kg (preferably 3 to 30 mg / kg) per adult and per day, and this dosage can be administered in one or several portions. When N-anisoyl-GABA or p-anisic acid was administered orally to rats, acute toxicity (LD50 values) of N-anisoyl-GABA was greater than . 000 mg / kg in both sexes, and that of p-anisic acid was 1.813 and 2.124 mg / kg in male and female respectively. When both drugs were repeatedly administered orally to rats over a period of 4 weeks, subacute toxicity, such as lethality and abnormality in haematological, haematobiochemical and toxipatological analyzes, was not observed up to 600 mg / kg. The potentiating effects of the release of ACh, by N-anisoyl-GABA and p-anisic acid, and the improving effects of circadian rhythm disorder and sleep disorder by 1-p-anisoyl-2-pyrrolidinone, are specifically described then .

Example 1 Test of the enhancing effect of ACh release Test animals: spontaneously hypertensive male rats prone to apoplexy (SHRSP) of 13 weeks of age, which received 1% solution of NaCl instead of water 5 days before the experiment.

Test method: The SHRSP were anesthetized and a guide cannula was implanted in the thalamus, dorsal hippocampus and prefrontal cortex. After recovery, a concentric microdialysis probe was inserted into the guide cannula and perfused with normal Ringer's solution containing 10 ~ 5 M of eserine (SIGMA, St. Louis, USA) with a constant flow rate of 2 μl. / min in conditions of freedom of movement. N-anisoyl-GABA or p-anisic acid were dissolved in Ringer's solution at final concentrations of 10"'', 10" 6 and 10"5 M, and each drug was perfused for 20 minutes through the same probe. The dialysates were collected every 20 minutes and injected into the high pressure liquid chromatography system to quantify the extracellular levels of ACh The release of ACh was expressed as the change in percent on the average of three consecutive stable samples collected before the perfusion of the drug.

Results of the trial: N-anisoyl-GABA potentiated the release of ACh by 32%, 48% and 70%, and p-anisic acid increased it by 22%, 51% and 61% in the thalamus (Fig. 1), dorsal hippocampus (figure 2) and prefrontal cortex (figure 3), respectively. In contrast, the effect of l-p-anisoyl-2-pyrrolididone was not observed in any of the brain regions. Therefore, it was assumed that N-anisoyl-GABA and p-anisic acid were active substances that contributed to the activation of central cholinergic neurons (potentiation of ACh release), as the main metabolites of lp-anisoyl-2 - pyrrolidione. Example 2 Test of the improving effect of the harmful circadian rhythm Test animals: Male Wistar rats of 9 weeks of age (young group) and around 30 months of age (old group). 0 Test method: The animals were individually stabilized and had free access to food and water. After a 24-hour fast (day 0), food was restricted to only 1 hour / day after 13.30 for 6 consecutive days. Food 5 was again denied on day 7, and the anticipated circadian activity motivated by food was investigated. L-p-anisoyl-2-pyrrolidinone at 30 and 100 mg / kg or vehicle was administered immediately after meal time, once a day or for 7 consecutive days. Spontaneous motor activity was measured in each cage.

•• *. Test results: In the old rats, the anticipated activity motivated by the food on day 5 7 was markedly attenuated compared to that of the young rats (figure 4), suggesting an abnormality in circadian rhythm regulation and a deficiency of the ability to adjust to the time due to aging. Repeated oral administration of l-p-anisoyl-2-pyrrolidinone, but not the vehicle, significantly improved the annoying activity anticipated in the older rats (Figure 5).

Example 3 Test of the improved effect of the clogged sleep model Test animals: SHRSP males of 13 weeks of age, which received a 1% solution of NaCl instead of water for 5 weeks, and Wistar Kyoto (WKY) rats of the same age.

Test method: The animals were anesthetized, electrodes were implanted for the electroencephalogram (EEG) and a sensor for brain temperature, inside the cerebral cortex, and electrodes were implanted for elelectromiogram (EMG) inside the muscle cervical back All the variables were recorded continuously for 7 days, and the behavioral states of the rats were classified as insomnia, REMS, NREMS analyzing the amplitude and frequency of EEG and EMG waves. The l-p-anisoyl-2-pyrrolidinone at 15 mg / kg or vehicle was administered orally twice a day (morning and evening) for 5 consecutive days.

Test results: Compared with the WKY control, SHRSP showed a reduction in REMS during the light period (sleep period), and an increase in NREMS and a decrease in brain temperature during the dark period (active period) , indicating a sleep-wake rhythm disorder (figure 6). Repeated oral administration of l-p-anisoyl-2-pyrrolidinone, but not from the vehicle, improved the reduced diurnity REMS in the SHRSP (Figure 7). Based on the above, it is concluded that N-anisoyl-GABA or p-anisic acid are useful as a remedy for several neuropsychiatric symptoms, such as circadian rhythm disorder, sleep disorder, attention deficit disorder and problematic behaviors (delirium and nocturnal walks), which are observed not only in cerebrovascular diseases (namely cerebral infarction and hemorrhage), but also in neuronal degeneration (namely, Alzheimer's disease, Parkinson's disease and progressive supranuclear palsy) and hyperkinetic syndrome (hyperactivity disorder and attention deficit). In addition, a combined effect of N-anisoyl-GABA and p-anisic acid, as well as each individual effect, can be expected sufficiently.

Example 4 Preparation of pharmaceutical compositions The pharmaceutically acceptable preparations described below are only suitable examples, but should not be considered in any way as limiting the pharmaceutical compositions of the present invention. .1 Preparation of a tablet containing N-anisoyl-GABA A tablet containing 100 mg of N-anisoyl-GABA was prepared by the following method, using the following compositions (per tablet) Composition A: N-anisoyl-GABA 1 0 0 mg Lactose 2 0 mg Kollidon CL (BASF) 1 5 mg Corn starch 3 0 mg Avicel PH 101 (Asahi Chemical Co., Ltd) 50 mg Composition B: Polyvinylpyrrolidinone K-90 5 mg Clear anhydrous silicic acid 18 mg Magnesium stearate 2 mg Total 240 mg A mixture of the composition A described above is kneaded in an 8% aqueous solution of polyvinylpyrrolidinone K-90. After drying at 60 ° C, the composition B is mixed with the previous one. The mixture is pressed into tablets to give a circular tablet with a weight of 240 mg and a diameter of 8 mm. 4. 2 Preparation of a tablet containing p-anisic acid A tablet containing 100 mg of p-anisic acid is prepared by the following method, using the following compositions (per tablet).

Composition A: p-anisic acid 100 mg Lactose -20 mg Kollidon CL (BASF) 15 mg Corn starch 30 mg Avicel PH 101 (Asahi Chemical Co., Ltd, 50mg Composition B: Polyvinylpyrrolidinone K-90 5 mg Light anhydrous silicic acid 18 mg Magnesium stearate 2 mg Total 240 mg A mixture of composition A described above was kneaded in an 8% aqueous solution of polyvinyl pyrrolidinone K-90. After drying at 60 ° C, composition B was mixed therewith. The mixture was pressed into circular tablets with a weight of 240 mg and a diameter of 8 mm. 4. 3 Preparation of a capsule containing N-anisoyl-GABA A capsule containing 100 mg of N-anisoli-GABA was prepared by the following method using the following compositions (per capsule).Composition A: N-anisoyl-GABA 1 00 mg Lactose 2 0 mg Kollidon CL (BASF) 2 mg Corn starch 53 mg Composition B: Polyvinylpyrrolidinone K-90 5 mg Avicel PH 101 (Asahi Chemicals Co., Ltd.) 18 mg Magnesium stearate 2 mg Total 200 mg A mixture of composition A described above was kneaded in an 8% aqueous solution of polyvinylpyrrolidinone K-90. After drying at 60 ° C, composition B was mixed therewith. The mixture was poured onto a gelatin capsule No. 3 to obtain a capsule containing 200 mg. It is noted that in relation to this date, the best method known to the applicant, to implement said invention is that which is clear from the manufacture of the objects to which it refers. Having described the invention as above, the content of the following is claimed as property.

Claims (6)

1. Use of N-anisoyl-GABA or p-anisic acid for the preparation of a pharmaceutical composition containing N-anisoyl-GABA or p-anisic acid for the enhancement of cerebral acetylcholine release.

2. Use of claim 1, wherein the pharmaceutical composition is for the treatment of circadian rhythm disorders, sleep disorders, attention deficit disorders and problem behaviors.

3. Use of claim 1 or claim 2, wherein the N-anisoyl-GABA or p-anisic acid is present as an active ingredient, in a unit dosage of 1 to 300 mg / kg per adult per day.

4. A pharmaceutical composition for the enhancement of cerebral acetylcholine release, characterized in that it contains N-anisoyl-GABA or p-anisic acid as an active ingredient, and a therapeutically inert filler.

5. The pharmaceutical composition as claimed in claim 4, characterized in that the treatment of circadian rhythm disorders, sleep disorders, attention deficit disorders and problematic behaviors.

6. The pharmaceutical composition according to claim 4 or claim 5, characterized in that the active ingredient N-anisoyl-GABA or p-anisic acid are administered in a dosage of 1 to 300 mg / kg per adult per day. AGENT TO IMPROVE THE LIBERATION OF CEREBRAL ACETILCOLINE SUMMARY OF THE INVENTION The present invention relates to the use of N-nisoyl-α-aminobutyl acid (N-anisoyl-GABA) or p-anisic acid, for the preparation of a pharmaceutical composition that enhances the release of acetylcholine (ACh) in brain. In particular, the invention relates to the use of N-anisoyl-GABA or p-anisic acid for the treatment of circadian rhythm disorders, sleep disorders, attention deficit disorders and problematic behaviors. The invention also relates to pharmaceutical compositions containing N-anisoyl-GABA or p-anisic acid as an effective ingredient.