MXPA98003432A - Treatment of traumat cerebral damage - Google Patents

Abstract

The compound 5- (2-ethyl-2H-tetrazol-5-yl) -1,2,3,6-tetrahydro-1-methylpyridine of the formula (I) improves cognitive functioning and attenuates neuron damage reductions cholinergic in traumatic brain injury models and is useful for the manufacture of a pharmaceutical preparation for the treatment of traumatic brain damage

Description

TREATMENT OF TRAUMATIC BRAIN DAMAGE FIELD OF THE INVENTION The present invention relates to the use of - (2-ethyl-2H-tetrazol-5-yl) -1, 2, 3, 'ß-tetrahydride-1-methylpyridine for the manufacture of a pharmaceutical preparation for the treatment of traumatic brain injury (TBI).

BACKGROUND OF THE INVENTION EP-A1-0 296 721 discloses a class of piperidine or 1, 2, β, 6-te t rahydropyridine substituted in the 5-position with a five-membered heterocyclic group including a subclass of 5-te razol il compounds. - 1, 2, 3, 6-optionally substituted rahydropyridine. The compounds were described as having high affinity to central cholinergic receptors, in particular high affinity to central muscarinic receptors, thus being useful in the treatment of Alzheimer's disease, senile dementia and impaired learning and memory functions.

The structure-activity relationship of this subclass was described by Moltzen et al., J. Med. Chem. 1994, 37, 4085-4099. One of the compounds, ie 5- (2-ethyl-2H-tetrazol-5-yl) -1,2,3,6-tetrahydro-1-methyl formamide, has been reported as selective for muscarinic receptors with much higher affinity for M1 than for M2 and M3 receptors (Sub types of Muscarin ic Recep tors, The Si x th In t erna ti ona l Sympos iu, Nov. 9-12, 1994, Fort La uderda). Functionally, it has been described that it behaves as a partial agonist at M? and an antagonist at M2 and M3 receptors. In addition, the only prominent effect reported was the effect on spatial memory acquisition in young and mature rats, respectively. The BIT caused by physical or neurological conditions or several diseases, is of great importance among the population and there is a great demand for effective and safe drugs for the treatment of such disorders and their sequelae. Now, it has surprisingly been found that the compound 5 - (2-yl-2H-tet-razol-5-yl) -1,2,3,6-tetrahydro-1-met ilpyridine shows beneficial effects in the treatment of TBI and its aftermath.

BRIEF DESCRIPTION OF THE INVENTION Accordingly, the present invention relates to the use of 5 - (2 - and il - 2H - tet razol - 5 - i 1) -1, 2, 3, 6, - t et rahidro-met ilf ormamide or an acid salt of addition of the same for the manufacture of a pharmaceutical preparation for the treatment of traumatic brain damage or its sequelae. The term traumatic brain injury (TBI) in this specification, represents that it includes all conditions associated with trauma to the brain or spinal cord, for example, caused by physical forces acting on the skull, or spinal cord, ischemia, shock, oppression when breathing, cardiac oppression, thrombosis or cerebral embolism, neurological problems caused by AIDS, cerebral hemorrhage, encephalomyelitis, hydrocephalus, post-operative cases, cerebral infections, contusions or elevated intracranial pressure. The pharmaceutically acceptable acid addition salts of the compounds used in the invention are formed with non-toxic organic or inorganic acids. Illustrative of such organic salts are those with maleic, fumaric, benzoic, ascorbic, pamoic, succinic, oxalic, bis-methyl ester, ionic, sulfuric, acetic, propionic, tartaric, salicylic, citric acid. , gluconic, lactic, malic, cmanic, citraconic, aspartic, stearic, palmitic, itaconic, glycolic, p-amindbenzoic, glucanic, -benzenesulfonic acid and theophylline, as well as the 8-haloteofyldins, for example 8-bromo-theof ilin. Illustrative of such inorganic salts are those with hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric and nitric acids. It has now been found that the compounds used according to the invention are useful in the treatment of TBI. In this way, for example, improves cognitive functioning followed by moderate traumatic brain damage and attenuates 5 reduced damage productions of cholinergic neurons. In addition, he has found that it does not cause cardiac effects or other adverse side effects in doses that are clinically relevant. In another aspect, the present invention provides a method for preventing or treating TBI in a human being comprising the step of administering a therapeutically effective amount of 5- (2-ethyl-2H-tet razol-5-yl) - 1, 2, 3, 6 -tet rahydro-1-met ilpyridine or an acid addition salt thereof to a patient in need of the same. The compound used according to the invention and the pharmaceutically acceptable addition salts thereof can be administered in any suitable form, for example orally or parenterally, and the compounds can be present in any suitable form for such administration, for example, in the form of tablets, capsules, powders, syrups or solutions or dispersions for injection. An effective daily dose of the compound according to the invention or a pharmaceutically acceptable salt thereof is from 10 μg / kg to 10 mg / kg of body weight, preferably from 25 μg / day / kg of body weight to 1.0 mg / day. / kg of body weight. Accordingly, an adequate daily dose is 500 μg to 600 mg / day, preferably 1.0 mg to 100 mg. The compound used according to the invention can be obtained as described in EP-A1 0 296 721 and its acid addition salts are readily prepared by methods well known in the art.

PHARMACOLOGY The compound used according to the invention was tested by the following well-suited and reliable method: * Cognitive function after TBI Rats were subjected to traumatic cerebral fluid percussion brain damage as described by Dixon, CE et al., J., Neurosurgery, 67 (1987) 110-119. The damaged animals were treated s.c. daily on days 1-15 after damage beginning 24 hours after damage with either saline or 5 - (2-ethyl-2H-tetrazol-5-yl) -1, 2, 3, 6-tetrahydro- l-methylpyridine, 3. 6 μmol / kg or 15 μmol / kg.

A correct reflex suppression was determined after TBU according to the method of Dixon et al 1997, supra. Body weight was recorded before damage and on days 1-5 after damage, and rotarodium functioning was determined on days 1-5 after damage according to the method of Hamm, RJ, et al., J. Neurotrauma, 11 (1994) 187-196. The rotarodio test was used to measure motor performance after TBI. Finally, the average latency (+ S.E.M.) Was measured to find the target platform in the Morris water maze on days 11-15 after damage. The results were analyzed through an ANOVA. During the water maze test, all animals were injected 10 minutes before the determination of the water maze. Simulatedly damaged animals (animals prepared for damage but without a pulse supply) were included for comparison.

How T i fi cation of Neonates of ChA T after TBI Rats were subjected to TBI of central fluid percussion and treated as described above. They were injected, s.c. with a saline solution (n = 5) or the test compound (5 - (2 -et i 1 - 2H -tetrazol-5-yl) -1,2,3,6-tetrahydro-l-methylpyridine, 15 μmol / kg) (n = 5). The simulated damaged rats were injected (s.c.) with a saline solution (n = 4) or the test compound 15 μmol / kg (n = 4). The straight reflex suppression was determined as described above. A possible loss of cholinergic neurons after TBI was determined through the quantification of acet ilt ransphrase of choline (ChAT) immuno - reactive neurons in. the basal forebrain On day 15 after the damage (2-4 hours after the last injection), all the animals were anesthetized with pentobarbi tal (90 mg / kg, ip) and were applied 'perfusion transartically, with 200-250 ml of an isotonic saline solution followed by 500 ml of 4.0% paraformaldehyde 0.2% picric acid in a pH regulator of 0.1 M phosphate at a rate of 500 ml / 30 minutes at room temperature. After perfusion, the brains were cut in two blocks and post-fixed for 24 hours in the same solution at 10 ° C. Sections of 40 μm coronal were collected in a vibratome through the nuclei of the anterior brain and each fifth section was processed for ChAT immunoreactivity. Parallel sections were stained for the Nissi substance with cresyl violet for the quantitative determination of any possible cholinergic or non-cholinergic neuronal loss in the basal forebrain nucleus. Sections of the free floating anterior brain were incubated in a final ChAT antibody (1:50) presentation of 1.0 μg / ml in a pH regulated saline solution of 0.01 M phosphate (PBS) containing 0.1% detriton X-lßO . Sections of the forebrain were cut and incubated in the primary antibody for 24 hours at room temperature in culture trays (4 sections / 300 μl / well). After 4 washes in PBS, the sections were incubated in the secondary antibody (horse anti-mouse IgG; Vector Laboratories, Burlingame, CA) for 1 hour at 37 ° C. After 3 washes in PBS, sections were incubated with a complex of avidin-biot mouse in-peroxidase (ABC) technique (Vector) (Hsu et al J. Histochem.

Cytochem. 29 1981, 577-580) for 2 hours at 37 ° C. After three washes with PBS and one in a saline solution regulated at its pH with 0.1M Tris (TBS), free flotation sections were processed through the oxidase-diaminobenzidine-nickel gucose method described by Shu et al. Neurosci. Lett. 85 1988, 169-171. The reaction was stopped by transferring the sections to TBS. The sections were mounted on glass slides coated with chromium-gelatin alumina and allowed to dry overnight at room temperature, dehydrated in previous concentrations of ethanol and xylene, and then covered with Permount. . The immunoreactive neurons of ChAT in the middle setal nucleus (MSN), the vertical limbus nucleus of the diagonal band (VDB) and the nucleus basalis "nucleus (NMB) were counted using an image formation device. Microcomputer (MCID) (Imaging Research Inc., Ontario, Canada) The limit between MSN and VDB was defined as the anterior commissure.The NMB was defined as the immunolabelled neurons located in the globulus pallidus and the junction of the internal capsule A cell count was made of each animal obtained at 0.2 mm intervals through the anterior brain nucleus Cell numbers were reported as group means per 10,000 μm2 for each nucleus of the anterior brain MSN neurons, VDB and NMB for parallel sections stained with cresyl violet were also counted in the MCID due to the large number of total cholinergic and non-cholinergic neurons that are stained with cresyl violet at Across these regions, a bilateral sample of neurons was counted in each nucleus. A grid with predetermined areas was used to count the cells for each nucleus. For the nuclei of MSN and VDB, the counting "of three regions of 2,000 μm2 on each side was made (in this way a total area of 12,000 μm2 was shown for each of the nuclei in each of the four sections represented. NMB a region of 12,000 μm2 was counted on each side (total area equal to 24,000 μm2 in each of the four sections counted). Only large cells (diameter> 20 μm) were quantified with visible somatic Nissi bodies and with a neural type transparent neural type nucleus.

Resul ates There were no significant differences between the animals damaged in the vertical reflex suppression after TBI. This indicates that within each experiment, the damage groups received an equivalent damage severity. The supposedly damaged animals were corrected significantly faster than some of the animals with TBI (p < .0001 for each comparison). The suppression of correction in the group allegedly damaged was due to gas anesthesia used before the damage. No significant difference was observed between any of the groups damaged in the loss of body weight after TBI, again indicating an equivalent damage severity among the damaged groups. There were no significant differences in the functioning of rotarodio on days 1-5 after damage between any of the damaged groups. Since the test compound was administered on days 1-15 after the damage, these tests also indicate that the drug had no effect on body weight or functioning of rotarodium after damage.

ANOVA indicated that the damaged animals injected (s.c.) daily with the test compound, worked better in the Morris water maze than the animals treated with a damaged saline solution. Damaged animals treated with the test compound 15 μmol / kg significantly improved performance with p < 0.01. The TBI caused a significant reduction in the number of ChAT neurons in VDB and NMB in rats treated with a saline solution and the test compound, respectively. However, the compound of the invention significantly attenuated reductions in damage reduced in ChAT-IR (32% "reduction compared to the test group treated with a saline solution bathed in VDB and 51% in NMB.) The sections stained with violet. of parallel cresyl did not show any reduction in the number of cells in MSN, VDB or NMB indicating that loss in neurons in ChAT-IR did not result in cell death.

Formulation Examples The pharmaceutical formulations of the invention can be prepared by conventional methods in the art.

For example: the tablets can be prepared by mixing the active ingredient with ordinary auxiliaries and / or diluents and subsequently containing the mixture in a conventional tablet-making machine. Examples of auxiliaries or diluents comprise: corn starch, lactose, talc, magnesium stearate, gelatin, lactose, gums and the like. Any other auxiliary or coloring additive, aroma, preservatives, etc. may be used, provided they are compatible with the active ingredients. The solutions for injection can be prepared by solvating the active ingredient and possible additives in a separate vehicle, preferably sterile water, adjusting the solution to the desired volume, sterilizing the solution and placing it in ampoules or flasks. Any suitable additive conventionally used in the art can be added, such as tonicity agents, preservatives, antioxidants, etc. Typical examples of preparations for the formulations of the invention are presented below (the amounts of the active ingredient are calculated as the free base): 1) Tablets: 5- (2-ethyl-2-tf-tetrazol-5-yl) -1, 2, 3, 6-tetrahydro-1-met ilpir idine 20 mg Lactose 60 mg Cornstarch . 30 mg hydroxypropylcellulose 2.4 mg Microcrystalline cellulose 19.2 mg Sodium croscarmellose type A 2.4 mg Magnesium stearate 0.84 mg 2) Tablets 5- (2-ethyl-2-tetra-zol-5-yl) - 1, 2, 3, 6-tetrahydro-l-methyl-pyridine 10 mg Lactose 46.9 mg Corn starch 23.5 mg Povidone '1.8 mg Microcrystalline cellulose 14.4 mg Croscaramellose Sodium Type A 1.8 mg Magnesium stearate 0.63 mg 3) Syrup: 5- (2-ethyl-2H-etrazol-5-yl) 1, 2, 3, 6-tetrahydro-l-methyl-pyridine 5.0 mg Sorbitol 500 mg Hydroxypropylcellulose 15 mg Glycerol 50 mg Met ilparaben 1 mg Propylparaben. 0.1 mg Ethanol 0.005 mi Taste 0.05 mg Natrium of saccharin 0.5 mg Water ad 1 mi 4) Solution 5- (2-ethyl-2H-tetrazol-5-yl) 1, 2, 3, 6-tetrahydro-l-methyl-pyridine 1.0 mg Sorbitol 5.1 mg Acetic acid 0.08 mg Water for injection ad 1 mi

Claims (4)

RESV? NPICACIONEIS

1. The use of 5- (2-ethyl-2H-tet razol-5-yl) 1, 2, 3, 6-tetrahydro-l-methylpyridine or a pharmaceutically acceptable acid addition salt thereof for the manufacture of a pharmaceutical preparation for the treatment of traumatic brain injury.

2. The use according to claim 1, characterized in that the manufactured pharmaceutical preparation comprises 5 - (2-ethyl-2H-tet razol-5-yl) -1, 2, 3, 6-tet ahydro-1 -met i pyridine or a pharmaceutically acceptable acid addition salt thereof in a unit dose form.

3. The use according to the claim 1, characterized in that the prepared pharmaceutical preparation manufactured comprises 5 - (2-ethyl-2H-tetrazol-5-i 1) -1,2,3,6-tetrahydro-1-met ilpyridine or a pharmaceutically acceptable acid addition salt thereof in an amount of 500 μg to 600 mg / day, preferably 1.0 mg to 100 mg / day.

4. The use according to any of claims 1-3, characterized in that the manufactured pharmaceutical preparation is for the treatment of traumatic brain damage caused by typical forces acting on the skull or spinal cord, ischemia, shock, oppression when breathing, attack cardiac, cerebral thrombosis or embolism, neurological problem caused by AIDS, cerebral hemorrhage, encephalomyelitis, hydrocephalus, post-operative cases, cerebral infections, contusions or elevated intercranial pressure and for the treatment of sequelae of such condition.