MXPA00010458A - Methods for increasing levels of acetylcholine - Google Patents

Abstract

The current invention relates to a method for increasing levels of acetylcholine in mammals comprising administering to a mammal in need thereof, an effective amount of a compound of formula (I), and optionally an AChE inhibitor.

Description

COMPOUNDS TO INCREASE THE ACETILCOLINE LEVELS The present invention deals with the disciplines of medical chemistry, neurophysiology and neuropharmacology. Specifically, the present invention relates to the increase of acetylcholine levels by the administration of 2-aryl-3-aroylbenzo [b] thiophenes. Cholinergic neurons constitute a major neuronal system of the central and peripheral nervous systems. Cholinergic neurons are especially associated with the neurotransmitter, acetylcholine. In the central nervous system, acetylcholine is a neurotransmitter and can be found, among other places, in the hippocampus and the frontal cortex of the brain. The hippocampal area of the brain, particularly those areas that are known to comprise cholinergic neurons, is thought to have functions associated with cognition, learning and memory. Degenerative diseases with symptoms such as loss of cognition, learning and memory, have been linked to Ref. 123888 a loss in colmergic neurons. For example, it is known that in patients suffering from Alzheimer's disease, there is a marked decrease in the level of cholinergic neurons in the hippocampus. The progressive loss of these colmérgicas neurons seems to reflect the progressive loss in the memory and the cognitive function in these patients. It is thought that one reason for the decline of these neurons is the loss or decreased function of the neurotransmitter, acetylcholine. Several potential therapies that are designed to increase acetylcholine levels are being clinically evaluated. The level of acetylcholine in a neuron is determined basically by where is the balance between its biosynthesis and bio-degradation. The enzyme colma-acetiltransferase (ChAT) is responsible mainly for its synthesis and acetylcholine esterase (AChE) for its degradation. A therapeutic strategy to increase the level of acetylcholine is based on the blocking of its degradation via the inhibition of AChE, for example, using AChE inhibitors such as physostigmine salicylate, tacrine hydrochloride, donepezil hydrochloride and the like. Although, there are some Encouraging results with the clinical use of AChE inhibitors, especially in the early stages of Alzheimer's disease, these agents generally have undesirable side effects due to their non-specific, systematic action. Currently, tacrine has been tested for the early treatment of Alzheimer's symptoms, (see "Goodman and Gilman's, The Pharmacology Basis of Therapeutics," Ed. Gilman, et al., Pergamon Press, 8th Ed., Chap. , (1990) and references cited therein Another therapeutic strategy to increase acetylcholine levels is based on the accelerated regulation of ChAT in neurons.The hormone, estrogen, has been found to increase the level of acetylcholine by rapidly regulating acetylcholine. ChAT in the hippocampus of rats (see "Immunochemical demonstration of increased choline acetyltransferase concentration in rat preoptic area after estradiol admmistration", Luine, et al., Brain Res., 191: 273-277, 1980, "Estradiol Increases Choline Acetyltransferase Activity "Specific Basal Forebram Nuclei and Projection Areas of Female Rats", Luine, V., Exp. Neurology, 89: 484-490, 1985, "Ovarían steroid deppvation results in reversible learning impairment and compromised cholinergic function in female Sprague-Dawley rats ", Singh, M., et al., Brain Res., 644: 305-312, 1994.) Therefore, it has been speculated, and Preliminary clinical information confirms that post-menopausal women treated with hormone replacement therapy (estrogen with or without progestins) may be less likely to succumb to Alzheimer's disease or have existing symptoms relieved. with estrogen has undesirable side effects, including uterotrophic effects, a possible increase in the incidence of breast cancer, swelling, resumption of menstruation, etc., which limits patient compliance. In this way, there is an opportunity for new and improved therapeutic interventions to increase acetylcholine levels. The present invention relates to a method for accelerating regulation of the colma-acetyl transferase (ChAT) in mammals, which comprises administering to a mammal in need therefor an effective amount of a compound of Formula I: or an acid, pharmaceutical or sorbate addition salt thereof; wherein: R1 and R3 are independently hydrogen, methyl, benzoyl, substituted benzoyl or C (0) - (C? -C6 alkyl); R2 is selected from the group of p? Rol? Dm-1-? Lo, p? Per? Dm-1-? Lo, and hexameth? Len? Mm-1-? Lo; where R2 is optionally the N-oxide; and optionally a colma-esterase inhibitor. In addition, the present invention relates to a method for increasing acetylcolma levels in the frontal cortex and / or hippocampal regions of the brain in mammals which comprises administering to a mammal in need thereof an effective amount of a compound of the Formula I, or an acid addition or solvate salt pharmacist thereof; and optionally a choline esterase inhibitor. Additionally, the present invention relates to a method for inhibiting the detrimental conditions or effects caused by a deficiency of choline-acetyltransferase and / or acetylcholine in the frontal cortex and / or hippocampal regions of the brain, in mammals, which comprises administering to a mammal in need thereof, an effective amount of a compound of Formula I, or a pharmaceutical salt or solvate thereof; and optionally a choline esterase inhibitor. In addition, the present invention relates to a pharmaceutical formulation comprising a compound of Formula I, or an acid addition salt or pharmaceutical solvate thereof, an acetylcholine esterase (AChE) inhibitor; and a pharmaceutical carrier, diluent or excipient. The present invention relates to the discovery that a select group of 2-aryl-3-aroylbenzo [b] thiophenes, ie, compounds of Formula I, are useful for accelerating the regulation of ChAT, and therefore are useful for increase acetylcholine levels in neurons that They contain acetylcholine and ChAT. A preferred embodiment of all the methods of the present invention is where the mammal to be administered with a compound of Formula I is a human, particularly a female human, more particularly when the female human is deficient in estrogen. However, male humans are also considered under the term "mammal" particularly male who are deficient in testosterone. Another preferred embodiment of the present invention is where the condition caused by a decrease in choline acetyltransferase and / or acetylcholine in the frontal cortex and / or hippocampal regions of the brain is Alzheimer's disease. In addition, another preferred embodiment of all methods of the present invention is the use of a pharmaceutical acid addition salt of a compound of Formula I, wherein R1 and R3 are hydrogen and R2 is p? Rol? D? N-1 Most preferably, the salt is the hydrochloride. This most preferred compound is called [2- (4-hydrox? Phen?) -6-h? Drox? Benzo [b] t? En -3? L] [4- [2- (l-p? rol? d? n? l) etox?] 'phen? l] methanone. Yet another more preferred embodiment of all the methods of the present invention is the use of a pharmaceutical acid, acid addition salt of a compound of Formula I, wherein R1 and R3 are hydrogen and R2 is p? Per? Dm-1- what? More preferably, the salt is the hydrochloride. This most preferred compound is called [2- (4-h? Drox? Phen?) -6-h? Drox? Benzo [b] t? En -3? L] [4- [2- (1) hydrochloride. -p? per? d? l) ethoxy] phenyl] methanone or "raloxifene hydrochloride." The present invention contemplates the optional use of commonly known AChE inhibitors such as igmma phosphate salicylate, tacrine hydrochloride, hydrochloride. of donepezil and the like, as well as agents that are ultimately found to be inhibitors of AChE As used herein, the term "effective amount" means an amount of a compound of Formula I that is capable of rapidly regulating ChAT and / or increase the levels of acetylcolma in the hippocampus and regions of the frontal cortex of the brain and / or inhibit the conditions or detrimental effects caused by a decrease in choline acetyltransferase and / or acetyl in mammals. When a compound of Formula I is co-administered with an AChE inhibitor, the term "effective amount" also means an amount of an agent that is capable of inhibiting AChE. The term "estrogen deficient" refers to a condition, whether it occurs naturally or is clinically induced, where a woman can not produce enough estrogen hormones to maintain estrogen-dependent functions, for example, menstruation, homeostasis bone mass, neuronal function, cardiovascular condition, etc. These deprived estrogen situations arise from, but are not limited to, menopause and surgical or chemical ovarectomy, including its functional equivalent, eg, medication with GnRH agonists or antagonists, ICI 182780, and the like. The term "inhibition" in the context of inhibiting the conditions or deleterious effects caused by a deficiency of ChAT and / or acetylcholine in the frontal cortex and / or regions of the brain's hippocampus includes its generally accepted meaning, ie, prohibition, restriction , relief, improvement, delay, stopping or reversing the progress or severity of a decrease in ChAT and acetylcholine and the pathological sequelae, that is, symptoms, that result from that event. The term "accelerated regulating ChAT" refers to the increase of the enzymatic activity of ChAT, that is, to promote the conversion of choline to acetylcholine. This promotion will include an increase in the efficiency and / or reaction rate of ChAT and choline and / or increase in the amount of ChAT present at the site of action. This increase in the amount of enzyme present may be due to the genetic regulation or another synthetic step of the formation of enzymes and / or a decrease in the deactivation and metabolism of the enzyme. The general terms used in the description of the compounds described in present have their usual meanings. For example, "Ci-Cβ alkyl" refers to straight, branched or cyclic aliphatic chains of 1 to 6 carbon atoms including methyl, ethyl, propyl, iso-propyl, cyclopropyl, n-butyl, pentyl, hexyl and the like. The term, "substituted benzoyl" is refers to a benzoyl group having one to five substituents independently selected from the C1-C4 alkyl, C1-C4 alkoxy, hydroxy, nitro, chloro, fluoro, or trifluoro or fluoro) methyl groups. The term "pharmaceutical" when used herein as an adjective means substantially non-toxic and substantially non-detrimental to the recipient. By "pharmaceutical formulation" is meant additionally that the carrier, solvent, excipients and salt must be compatible with the active ingredient of the formulation (a compound of Formula I). The term "acid addition salt" refers to a salt of a compound of Formula I prepared by the reaction of a compound of Formula I with a mineral or organic acid. For the use of pharmaceutical acid addition salts, see for example Berge, S.M., Bighley, L.D., and Monkhouse, D.C., J. Pharm. Sci., 66: 1, 1977. The term "solvate" represents an aggregate comprising one or more molecules of the solute, such as a compound of Formula I, with one or more molecules of a pharmaceutical solvent, such as water, ethanol and the like. The compounds of Formula I, wherein R and / or R3 are hydrogen or methyl can be prepared according to known procedures, such as those detailed in U.S. Patent Nos. 4,133,814; 4,418,068 and 5,731,342, the teachings of each are incorporated herein by reference. The compounds of Formula I which are carboxylic esters (R1 and / or R3 are 'C (O) - (Ci-Cβ alkyl), benzoyl, or substituted benzoyl) can be prepared from the compounds of Formula I where R and / or R3 are hydrogen by methods described in U.S. Patent No. 5,393,763, the teachings of which are incorporated herein by reference. The pharmaceutical acid addition salts of the invention are typically formed by reacting a compound of Formula I with an equimolar amount or an excess of acid. The reagents are generally combined in a mutual solvent such as diethylether, tetrahydrofuran, methanol, ethanol, isopropanol, benzene and the like. The salts normally precipitate from the solution in the space of approximately one hour to about ten days and can be isolated by filtration or other conventional methods. The acids commonly used to form acid addition salts are inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and the like, and organic acids such as p-toluenesulfonic acid, methanesulfonic acid, ethansulonic acid , oxalic acid, p-bromofemlsulphonic acid, carbonic acid, "succic acid, citric acid, tartaric acid, benzoic acid, acetic acid and the like.Fosstigma salicylate, tacpna hydrochloride, donepezil hydrochloride and other AChE inhibitors are commercially available. Pharmaceutical formulations can be prepared by methods known in the art such as, for example, in the published application EP 670162A1, published on September 6, 1995, and in WO 97/35571 published on October 2, 1997, both which are incorporated herein by reference, for example, a compound of Formula I, and optionally In an AChE inhibitor, can be formulated with common excipients, diluents or carriers, and tablets, capsules and the like are formed. In this manner, a compound of Formula I and an AChE inhibitor can be formulated or administered together. A compound of Formula I and an AChE inhibitor can also be administered separately. Examples of excipients, diluents and carriers that are suitable for the formulation include the following fillers and extenders such as starches, sugars, mannitol and salicylic derivatives; binding agents such as caboxymethyl cellulose and other cellulose derivatives, alginates, gelatin and polyvinyl pyrrolidone; wetting agents such as glycerol; disintegrating agents such as agar, calcium carbonate and sodium bicarbonate; agents for delaying dissolution such as paraffin; resorption accelerators such as quaternary ammonium compounds; surface active agents such as cetyl alcohol, glycerol monostearate; absorptive carriers such as caolma and bentonira; and lubricants such as talc, calcium and magnesium stearate and solid polyethylene glycols. The final pharmaceutical forms can be pills, tablets, powders, pills, syrups, aerosols, sachets, bottles, elixirs, suspensions, emulsions, ointments, suppositories, sterile injectable solutions, or sterile packaged powders, depending on the type of excipient used. Additionally, the compounds of Formula I are well suited for formulation as sustained release dosage forms. The formulations may also be constituted so that they release the active ingredient alone or preferentially in a particular part of the intestinal tract, possibly over a period of time. These formulations will comprise coatings, envelopes or protective matrices which can be prepared from polymeric substances or waxes. The particular dose of a compound of Formula I required to accelerate ChAT regulation, and optionally the dose of the AChE inhibitor required to inhibit AChE, according to this invention, will depend on the particular circumstances of the conditions being treated. Considerations such as dosage, route of administration and frequency of dosing are they decide better by the attending physician. In general, a minimum effective aosis for oral or parenteral administration of a compound of Formula I is about 1, 5, 10, 15, or 20 mg. Typically, an effective maximum dose is about 800, 100, 60, 50 or 40 mg. A particularly effective amount is 60 mg of raloxifene hydrochloride (56 mg of free base) per day via an oral route of administration. These doses will be administered to a patient in need of treatment from one to three times each day or as often as needed to effectively speed up ChAT, and / or increasing acetylcholine levels in the frontal cortex and / or hippocampal regions of the brain and / or to inhibit detrimental conditions or effects caused by a deficiency of colma-acetyltransferase and / or acetylcholine in the frontal cortex and / or regions of the brain hippocampus The following formulations are given for purposes of illustration and not to limit in any way. The total active ingredients in these formulations comprise from 0.1% to 99.9% by weight of the formulation. The term "ingredient "active" means a compound of Formula I, or a pharmaceutical salt or solvate thereof (preferably raloxifene hydrochloride) and optionally an AChE inhibitor.An even more preferred formulation of a compound of Formula I will be raloxifene hydrochloride in the form particular crystalline, particular size and composition illustrated in U.S. Patent No. 5,731,327 and PCT application WO 97/35571 (October 2, 1997), the teachings of which are each incorporated by reference.

Formulation 1 Gelatin Capsules Ingredient Quantity (mg / capsule) Active ingredient 50-600 Starch NF 0-500 Fluid powder of starch 0-500 Silicone fluid of 350 centistrokes 0-15 The ingredients are mixed, passed through a No. 45 mesh American sieve, and filled into hard gelatin capsules.

Formulation 2 Tablets Ingredient Quantity (mg / tablet) Active ingredient 50-600 Starch 10-50 Microcrystalline cellulose 10-20 Polyvinylpyrrolidone 5 (as a 10% solution in water) Sodium carboxymethylcellulose 5 Magnesium stearate 1 Talc 1-5 The active ingredient, starch and cellulose are passed through a North American mesh screen number 45 and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resulting powders which are then passed through a North American sieve of mesh number 14. The granules produced in this way are dried at 50-60 ° C and passed through a North American mesh screen. No. 18. The carboxymethylcellulose sodium, magnesium stearate and talc, previously based through a North American mesh sieve number 60, are added to the previous granules and they mix completely. The resulting material is compressed in a tabletting machine to produce the tablets.

Formulation 3 Aerosol Ingredient% by weight Active ingredient 0.50 Ethanol 29.50 Propellant 22 70.00 (Chlorodifluoromethane) The active ingredient is mixed with ethanol and the mixture is added to a portion of the propellant 22, cooled to -30 ° C and transferred to a filling device. The required amount is then fed to a stainless steel container and diluted with the rest of the propellant. The valve units are then equipped to the vessel.

Formulation 4 Suspension Ingredient Weight / volume Active ingredient 100 mg Sodium carboxymethylcellulose 50 mg Syrup 1. . 25 my Benzoic acid solution (0.1 M) 0. . 10 my Taste c. , v. C color . , V. Purified water total 5 mi Suspensions each containing 100 mg of a compound of Formula I per 5 ml of doses are prepared as follows, the active ingredient is passed through a No. 45 mesh American sieve and mixed with the sodium carboxymethyl cellulose and syrup for form a soft paste. The benzoic acid solution, the taste and the color diluted in water are added and the mixture is completely stirred. The additional water is added to bring the complete mixture to the required volume.

The following demonstration of the methods of the present invention is presented for purposes of illustration and is not intended to limit the scope of this invention in any way. Forty female Sprague-Dawley rats (Weight range of 300 to 325 g, six months old) are obtained from Harán. The animals either undergo bilateral ovariectomy (OVX) or are exposed to a Sham surgical procedure, and then shipped after two weeks. "Upon arrival, they are housed in metal hanging cages in groups of 3 or 4. per cage and have ad libitum access to food and water for a week.The ambient temperature is maintained at 22.2 ° ± 1.7 ° C with a minimum relative humidity of 40% The photopepod in the room is 12 hours of light and 12 hours The animals are dosed daily by subcutaneous injection or oral priming with either raloxifene hydrochloride at 3 mg / kg / day in a vehicle containing 10% cyclodextrin, estradiol benzoate at 0.03 or 0.3 mg / kg / day. day, or a vehicle control.The animals were treated for 3 to 10 days.There were twenty animals for each dosing regimen. appropriate time, the animals are sacrificed and the brains are extracted. The particular portions of the brains are homogenized and valued. The homogenates of the hippocampus and frontal cortex are processed and the activity of ChAT activity is determined by a radio-labeled assay of acetylcholine bio-synthesis. This procedure can be found in Schoepp et al., J. Neural Transmiss., 78: 183-193, 1989, the teachings of which are incorporated "as a reference." As expected, in animals with OVX, the levels of ChAT they are reduced to> 50% (p <0.001) compared to controls operated with sham In contrast, animals that received raloxifene hydrochloride or estradiol benzoate have significantly increased levels (p <0.05) of ChAT against OVX controls and a more significant difference of the sham controls In this way, the present invention provides methods for the treatment and prophylaxis of syndromes related to memory loss, learning and cognitive function, often seen in the woman who is deprived of estrogen, especially the post-menopausal woman. An example of this syndrome is senile dementia of the Alzheimer's type. The beneficial effects, such as the decrease in memory loss, associated with the administration of a compound of this invention become apparent after chronic administration. For example, the post-menopausal woman suffering from Alzheimer's disease can expect in demonstrating an improvement of his disease after 2-12 months of administration of raloxifene hydrochloride at 60 mg per day, via the oral route. The methods of the present invention can also be used in a prophylactic modality. For example, a group of peri- or post-menopausal women can have their memory cognitive function assessed by normal tests. After establishment of this baseline, women are administered raloxifene hydrochloride at 60 mg / day, via the oral route, for a period of 1-5 years. At the end of this time, the re-evaluation of the cognitive and memory functions by the normal tests shows a decrease in the loss of these functions in relation to a coupled set of patients. who have been given a placebo during the same period of time.

It is noted that in relation to this date, the best method known by the applicant to carry out the present invention is that which is clear from the present description of the invention. Having described the invention as above, the content of the following is claimed as property:

Claims (18)

1. The use of a compound of Formula I I; or a pharmaceutical acid or solvate addition salt thereof; wherein: R1 and R3 are independently hydrogen, methyl, benzoyl, substituted benzoyl or C (0) - (C? -C6 alkyl); R2 is selected from the group of pyrolidin-1-yl, piper idin-1-yl, and hexamethyleneimin-1-yl; where R2 is optionally the N-oxide; and optionally an acetylcholine esterase inhibitor (AChE); in the preparation of a useful drug to accelerate the regulation of choline-acetyltransferase (ChAT) in mammals.

2. The use of a compound of Formula I: I; or a pharmaceutical acid or solvate addition salt thereof; wherein: R1 and R3 are independently hydrogen, methyl, benzoyl, substituted benzoyl or C (0) - (C? -C6 alkyl); R2 is selected from the group of p? Rol? D? N-1-lio, piper? D? N-1-? Lo, and hexameth? Len? M? N-1-? Lo; where R2 is optionally the N-oxide; and optionally an acetylcholine esterase inhibitor (AChE), in the preparation of a medicament useful for increasing acetylcholine levels in the frontal cortex and / or hippocampal regions of the brain in mammals.

3. The use of a compound of Formula I: I; or a pharmaceutical acid or solvate addition salt thereof; wherein: R1 and R3 are independently hydrogen, methyl, benzoyl, substituted benzoyl or C (0) - (C? -C6 alkyl); R2 is selected from the group of p? Rol? D? N-1-? Lo, p? Per? D? N-1-? Lo, and hexameth? Len? M? N-1-? Lo; where R2 is optionally the N-oxide; and optionally an acetylchol esterase (AChE) inhibitor, in the preparation of a medicament useful for inhibiting the conditions or deleterious effects caused by a deficiency of choline acetyltransferase and / or acetylcholine in the frontal cortex and / or regions of the hippocampus. of the brain in mammals.

4. The use according to any of claims 1-3 wherein the mammal is a female human.

5. The use according to claim 4, wherein the female human is deficient in estrogens.

6. The use according to claim 5, wherein the compound of the Formula I is an acid addition salt, pharmaceutical, R1 and R3 are hydrogen, and R2 is p? Per? D? N-1-? Lo.

7. The use according to claim 6, wherein the compound of Formula I is the hydrochloride salt.

8. The use according to claim 5, wherein the compound of Formula I is a pharmaceutical acid addition salt, R1 and R3 are hydrogen, and R2 is p? Rol? D? N-1-? Lo.

9. The use according to claim 8, wherein the compound of Formula I is the hydrochloride salt.

The use according to claim 3, wherein the mammal is a human and the inhibited condition is Alzheimer's disease.

11. The use according to claim 10, wherein the human is a woman deficient in estrogen.

The use according to claim 11, wherein the compound of the Formula I is a pharmaceutical salt of acid addition, R1 and R3 are hydrogen, R2 is piper idin-1-yl.

13. The use according to claim 12, wherein the compound of Formula I is the hydrochloride salt.

The use according to any of claims 2 to 3, wherein the acetylcholine esterase inhibitor (AChE) is selected from physyme salicylate igmina, tacrine hydrochloride, and donepezil hydrochloride.

15. A pharmaceutical formulation characterized in that it comprises a compound of Formula I: I; or a pharmaceutical acid or solvate addition salt thereof; where: R1 and R3 are independently hydrogen, methyl, benzoyl, substituted benzoyl or C (O) - (Ci-Cβ) I rent); R2 is selected from the group of p? Rol? D? N-1-? Lo, p? Per? Dm-1-? Lo, and hexamet? Len? M? N-1-? Lo; where R2 is optionally the N-oxide; an acetylcholine esterase inhibitor (AChE); and a pharmaceutical carrier, diluent or excipient.

16. The formulation according to claim 15, characterized in that the compound of Formula I is a pharmaceutical acid addition salt, R1 and R3 are hydrogen, and R2 is p? Pepdm-1-? Lo.

17. The formulation according to claim 16, characterized in that the compound of Formula I is the hydrochloride salt.

18. The formulation according to rei indication 17, characterized in that the acetylcholine esterase inhibitor (AChE) is selected from physostigmine salicylate, tacrma hydrochloride and donepezil hydrochloride.