CN1207039A - Traumatic Brain Injury Treatment - Google Patents

Abstract

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

Description

The treatment of traumatic brain injury

Invention field:

The present invention relates to 5-(2-ethyl-2H-tetrazolium-5-yl)-1,2,3,6-tetrahydrochysene-1-picoline is used for the treatment of application in the pharmaceutical preparation of traumatic brain injury (TBI) in preparation.

Background of invention:

EP-A10 296 721 discloses a class piperidines or 1,2,3, the 6-5,6-tetrahydropyridine compounds, and its 5-position is replaced by a 5-unit heterocyclic radical, comprises one group of 5-tetrazole radical-1,2,3 that replaces arbitrarily, the 6-5,6-tetrahydropyridine compounds.Disclosed this compounds centering pivot cholinoceptor and had high-affinity, particularly to maincenter muscarine M ₁Receptor has high-affinity, therefore, can be used for treating Alzheimer, senile dementia, and learning and memory function barrier.

People J.Med.Chem.1994 such as Moltzen, 37,4085-4099 has described the structure-activity relation of this group of chemical compound.Reported wherein a kind of chemical compound, i.e. 5-(2-ethyl-2H-tetrazolium-5-yl)-1,2,3,6-tetrahydrochysene-1-picoline has selectively acting to M-ChR, and to M ₁Receptor comparison M ₂And M ₃Receptor have high several times affinity (hypotype of M-ChR, the 6th international symposium, Nov.9-12,1994, FortLauderdale).Its function is described to M ₁The partial agonist of receptor, and M ₂And M ₃The antagonist of receptor.And unique outstanding effect is the effectiveness that respectively young rat and adult rat spatial memory is obtained in the in vivo test of having reported.

In the crowd by physics or neural state, the perhaps caused TBI of various diseases, its importance constantly increases, therefore, to be used for the treatment of this disease and sequela thereof effectively and safe drugs huge demand is arranged.

Be surprised to find now, chemical compound 5-(2-ethyl-2H-tetrazolium-5-yl)-1,2,3,6-tetrahydrochysene-1-picoline demonstrates useful effect in being used for the treatment of TBI and sequela thereof.

Summary of the invention:

Therefore, the present invention relates to 5-(2-ethyl-2H-tetrazolium-5-yl)-1,2,3,6-tetrahydrochysene-1-picoline, or its acid-addition salts is used for the treatment of application in the pharmaceutical preparation of traumatic brain injury or its sequela in preparation.

In description and claims, noun " traumatic brain injury (TBI) " means and comprises all diseases relevant with brain or spinal cord injuries receptor, for example, by the physical force that acts on (Scull) at the bottom of the brain or spinal column, ischemia, apoplexy, respiratory arrest, heart beating stops, cerebral thrombosis or thromboembolism, neurological disorder due to the AIDS, cerebral hemorrhage, encephalomyelitis, hydrocephalus, the postoperative incident, brain infects, concussion or the caused disease relevant with brain or spinal cord injuries receptor of intracranial pressure rising.

The pharmaceutically useful acid-addition salts of this chemical compound that the present invention is used is the salt that forms with nontoxic organic or inorganic acid.The salt that the example of this class organic salt has following organic acid to form: maleic acid, fumaric acid, benzoic acid, ascorbic acid, pamoic acid, succinic acid, oxalic acid, two-the methylene salicylic acid, methanesulfonic acid, ethionic acid, acetic acid, propanoic acid, tartaric acid, salicylic acid, citric acid, gluconic acid, lactic acid, malic acid, mandelic acid, cinnamic acid, citraconic acid, chi winter propylhomoserin, stearic acid, Palmic acid, itaconic acid, glycolic, Para-Aminobenzoic, glutamic acid, benzenesulfonic acid, with theophylline acetic acid, and 8-halo theophylline, as 8-bromo theophylline.The example of this class inorganic salt has with hydrochloric acid, hydrobromic acid, sulphuric acid, sulfamic acid, the salt that phosphoric acid and nitric acid form.

Find that now the used chemical compound of the present invention is useful to treatment TBI.For example, it can improve the identification ability after the moderate traumatic brain injury, and the damage degenerative minimizing that can reduce cholinergic neuron.But also find that be considered under the clinical suitable dosage, it does not cause deleterious heartbeat or other side effect.

On the other hand, the invention provides a kind of method that is used to prevent or treat people TBI, comprise that the patient to this treatment of needs treats effective dose 5-(2-ethyl-2H-tetrazolium-5-yl)-1,2,3, the step of 6-tetrahydrochysene-1-picoline or its acid-addition salts.

Chemical compound that the present invention is used and pharmaceutically useful acid-addition salts thereof, can be by any suitable by way of administration, for example oral or parenteral administration, and, this chemical compound can also be with any suitable form administration, for example with tablet, and capsule, the form of powder or syrup, the perhaps injection type of solution or dispersion.

The compounds of this invention or its pharmaceutically useful salt, its effective daily dose are 10 μ g/kg body weight-10mg/kg body weight, preferably 25 μ g/ day/kg body weight-1.0mg/ day/kg body weight.Therefore, the daily dose that is fit to is 500 μ g/ day-600mg/ day, preferably 1.0mg/ day-100mg/ day.

The used chemical compound of the present invention can obtain according to the method described in the EP-A10 296 721, and its acid-addition salts can easily prepare by method well known in the art.The pharmacology

By following generally acknowledged reliable method, the chemical compound that the present invention is used is measured:

Recognition function after the TBI

According to people J.Neurosurgery such as Dixon.C.E, the described method of 67 (1987) 110-119 makes rat form maincenter fluid impact traumatic brain injury.To the animal of damaged from injured back 24 hours begin treatments, with normal saline or 5-(2-ethyl-2H-tetrazolium-5-yl)-1,2,3,6-tetrahydrochysene-1-picoline, with the dosage of 3.6 μ mol/kg or 15 μ mol/kg, every day, the subcutaneous injection administration was 1-15 days.

According to Dixon et al 1987, the righting reflex that the method for the same document is measured behind the animal TBI suppresses.

In 1-5 days record body weight before the animal damaged and after the damaged, and according to people J.Neurotreuma such as Hamm.R.J., the method among 11 (1994) 187-196 was measured the ability of animal swingle in 1-5 days in injured back.Swingle test can be used for measuring the motor capacity behind the TBI.

At last, in injured the last 11-15 days, measure animal in the Morris water maze, find the mean consumption time of target platform (+S.E.M.).By means of ANOVA this result is analyzed.In the water maze test process, all animals are measured preceding 10 minutes at water maze carried out injection for curing.

Also comprise in the test and be used for correlated false damage animal (make the animal damaged, but be not subjected to the maincenter fluid impact).

TBI is afterwards to the neuronic quantitative assay of ChAT:

Make rat form maincenter fluid impact traumatic brain injury as mentioned above, and treat.(n=5) animal is made subcutaneous injection with normal saline (n=5) or testing compound (5-(2-ethyl-2H-tetrazolium-5-yl)-1,2,3,6-tetrahydrochysene-1-picoline, 15 μ mol/kg).False injury rats is also treated with normal saline (n=4) or testing compound 15 μ mol/kg (n=4) injections (subcutaneous).

Measure the inhibition of righting reflex as mentioned above.

By forebrain basilar part choline acetyltransterase (ChAT) immunoreactivity neuron is carried out quantitative counting, can measure the TBI possible loss of cholinergic neuron afterwards.In 15 days (2-4 hour behind the drug administration by injection for the last time) after the damaged, with all animals with pentobarbital anesthesia (90mg/kg peritoneal injection) after, use earlier the 200-250ml isotonic saline solution, subsequently in order to 500ml 4.0% paraformaldehyde/0.2% picric acid of 0.1M phosphate buffer preparation, at room temperature, make the aorta perfusion with 500ml/30 minute speed.After perfusion is finished, choose and be trimmed to two cerebral tissue, and in identical solution, fix 24 hours after 10 ℃.By each nucleus of forebrain, on vibratome, cut the thick crown section of 40 μ m, and the ChAT immunoreactivity is made in the 5th section of each nuclear handled.Parallel section is done the dyeing of Nissi body material with crystal violet, be used for the neuronic any possible loss of quantitative assay forebrain basilar part nucleus cholinergic and non-cholinergic.

Preceding brain section is hatched as free-floating in the final ChAT antibody concentration of preparing with the 0.01M phosphate buffered saline (PBS) (PBS) that contains 0.1%triton, X-100 of 1.0 μ g/ml (1: 50).Then preceding brain section is repaired, and in culture plate, at room temperature hatched 24 hours (4/300 μ l/ hole) with first antibody.

After in PBS, washing 4 times, with these the section second antibody (the anti-mice IgG of horse, VectorLaboratories, Burlingame, CA) in 37 ℃ hatched 1 hour.After in PBS, washing 3 times, with mice affinity element-avidin-biotin complex (ABC) (Vector) technology (Hsu et al, J.Histochem.Cytochem29,1981,577-580) will cut into slices and hatch 2 hours at 37 ℃.Wash 3 times with PBS, wash after 1 time with 0.1M Tris buffer saline (TBI), by means of Shu et al, Neurosci.Lert.85 1988, the described glucoseoxidase-diaminobenzidine of 169-171-nickel method, the section of free-floating handled.By section is transferred to reaction is stopped.Section shop is mounted on the microscope slide of gelatin-chrome alum chromalum coating, and make it dried overnight at room temperature, in the ethanol of progressive concentration and dimethylbenzene, dewater, make the coverslip mounting with Permount then.

With microcomputer image processing system (MCID) (Imaging Research Inc.Ontario.Canada), to septal nucleus (MSN), the ChAT immunoreactivity neuron in diagonal band top nuclear (VDB) and the outside basal nuclei is counted.Interface between MSN and the VDB is defined as anterior commissure.NMB is defined as being arranged in the immune labeled neuron of HUANGBAI(sic) ball and capsula interna adjacency section.From every animal, cell counting is carried out in the section that obtains at interval by each nucleus 0.2mm of forebrain.For nuclei of cranial nerves before each, cell number is with per 10000 μ m ²Class mean represent.

For parallel slices with violet staining, its MSN, the neuron among VDB and the NMB is also counted by MCID.Because in whole cholinergics and non-cholinergic neuron, whole these zones are a lot of by the neuronal quantity of violet staining, so, each nucleus is carried out bilateral sample neuron count.The cell counting that area and netted grid chi are used for each nucleus will be pre-determined.For MSN and VDB nuclear, to 3 2000 μ m of every side ²The zone count (like this, every section in 4 sections that are counted, the gross area of the sampling of each nucleus sampling is 12000 μ m ²).For NMB, one 12000 μ m of every side counting ²Zone (every section in 4 sections that are counted, the gross area of counting=24000 μ m ²).Only to having obvious somatic cell Nissi body, and the maxicell (diameter＞20 μ m) with very typical neuronal cell nuclear is done quantitative assay.The result

Behind the TBI, between any animal of damaged, the inhibition of righting reflex does not have significant difference.This shows that in each experiment, the damage treated animal has been subjected to the damage of equal extent.False damage animal right action significantly faster than any animal of TBI (each relatively in P＜0.0001).The inhibition of false damage treated animal righting reflex is because the preceding used gas anesthesia of damage causes.

Between any animal of injured group, weight loss does not have significant difference behind the TBI, shows that once more injured treated animal has equal degree of injury.

Between any animal of damaged group, the ability of 1-5 days swingles does not have significant difference after damaged.Because be after damage, to give testing compound in 1-15 days, so these tests also show body weight or the swingle ability not influence of this medicine after to damage.

ANOVA the analysis showed that every day, the damaged animal than the pump pickle treatment in the Morris water maze had better performance with the damaged animal of testing compound injection (subcutaneous) treatment.Damaged animal with testing compound 15 μ mol/kg treatment has the ability of remarkable improvement, P＜0.01.

With the rat of saline and testing compound treatment, TBI has caused that ChAT-1R god significantly reduces with first quantity among VDB and the NMB for respectively.But The compounds of this invention has reduced the neuronic damage degenerative of ChAT-1R significantly and has reduced (among the VDB, compare with the damaged group of brine treatment, reduced by 32%, be reduced to 51% among the NMB).The parallel slices of violet staining shows, MSN, and cell number does not reduce among VDB or the NMB, shows that the neuronic loss of ChAT-1R is not because due to the cell death.

Example of formulations

Pharmaceutical preparation of the present invention can be by the method preparation of this area routine.

For example: tablet can be by with active component and common adjuvant and/or mixing diluents, then in the tablet machine of using always with this mixture tabletting.The example of adjuvant or diluent comprises: corn starch, lactose, Pulvis Talci, magnesium stearate, gelatin, natural gum or the like.Any other adjuvant or the pigment of interpolation, spice, antiseptic etc. are as long as they and this active component are compatiblely all can add.

Injection can prepare by following program: earlier active component and possible additive are dissolved in the part excipient, preferred excipient is a sterilized water, and this solution calibrated to required volume, again with this solution sterilization, be filled into then in the suitable ampoule or medicine bottle.Can also add the conventional any suitable additive that uses in this area, as osmotic pressure regulator, antiseptic, antioxidant etc.

The exemplary formulations example of preparation of the present invention is as follows: (amount of active component is to calculate with free alkali) 1) tablet: 5-(2-ethyl-2H-tetrazolium-5-yl) 1,2,3,6-tetrahydrochysene-1 20mg-picoline lactose 60mg corn starch 30mg hydroxypropyl cellulose 2.4mg microcrystalline Cellulose 19.2mgA type Croscarmellose sodium 2.4mg magnesium stearate 0.84mg2) tablet: 5-(2-ethyl-2H-tetrazolium-5-yl) 1,2,3,6-tetrahydrochysene-1 10mg-picoline lactose 46.9mg corn starch 23.5mg polyvinyl pyrrolidone 1.8mg microcrystalline Cellulose 14.4mgA type Croscarmellose sodium 1.8mg magnesium stearate 0.63mg3) syrup: 5-(2-ethyl-2H-tetrazolium-5-yl) 1,2,3,6-tetrahydrochysene-1 5.0mg-picoline sorbitol 500mg hydroxypropyl cellulose 15mg glycerol 50mg nipagin 1mg propyl parabene 0.1mg ethanol 0.005ml aromatic 0.05mg saccharin sodium 0.5mg water adds to 1ml4) solution 5-(2-ethyl-2H-tetrazolium-5-yl) 1,2,3,6-tetrahydrochysene-1 1.0mg-picoline sorbitol 5.1mg acetic acid 0.08mg water for injection adds to 1ml

Claims (4)

1. 5-(2-ethyl-2H-tetrazol-5-yl)-1,2,3,6-tetrahydro-1-picoline or its pharmaceutically acceptable acid addition salt is used in the preparation Application of pharmaceuticals in the treatment of traumatic brain injury.

2. The application according to claim 1, characterized in that the prepared medicament contains 5-(2-ethyl-2H-tetrazol-5-yl)-1,2,3,6-tetrahydro- 1-picoline or a pharmaceutically acceptable acid addition salt thereof. 3. The application according to claim 1, characterized in that the prepared medicament contains 500 μg-600mg/day, preferably 1.0mg-100mg/day dose of 5-(2-ethyl-2H-tetrazol-5-yl )-1,2,3,6-tetrahydro-1-picoline or a pharmaceutically acceptable acid addition salt thereof. 4. According to the application of any one of claims 1-3, it is characterized in that the prepared medicament is used for the treatment of physical force acting on the base of the brain or spine, local ischemia, stroke, respiratory arrest, cardiac arrest, cerebral palsy, etc. Thrombosis or embolism, neurological disorders due to AIDS, cerebral hemorrhage, encephalomyelitis, hydrocephalus, postoperative events, traumatic brain injury due to brain infection, concussion, or increased intracranial pressure, and/or for treatment Sequelae of such a disease.