GB2414665A - Aminoketone derivatives of phloroglucinol for the treatment of motor neuron disease - Google Patents

Abstract

Aminoketone derivatives of phloroglucinol are outlined for use in the preparation of medicaments for the treatment of motor neuron disease, including Amyotrophic Materal Sclerosis (AMS), Primary Lateral Sclerosis (PLS) and Progressive Muscular Atrophy (PMA) and the associated motor neuron degeneration and loss. The synergic combination of such an aminoketone derivative of phloroglucinol and magnesium and its pharmaceutically acceptable salts such as magnesium oxide, magnesium aspartate, magnesium sulphate, magnesium citrate, magnesium chelates, and magnesium EPA is also outlined, as well as the use of specific compounds bound to a cation-exchange resin, in the preparation of medicaments for treating motor neuron disease and associated motor neuron loss. The aminoketone derivatives of phloroglucinol outlined all have a 2,4,6-trialkoxyphenyl group bound to an aminoketone chain, with the amine being either pyrrolidinyl, piperazinyl, piperidinyl or a diethylamino group.

Description

24 1 4665

MOTOR NEURON LOSS

FIELD OF THE INVENTION

s Present invention is directed to a novel medicament for medical treatment of a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) spinal-bulbar muscular atrophy (SBMA, or Kennedy's disease). More particularly it involves the use of aminoketones derivatives of fluoroglucinol, to prevent, retard and a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) and the associated loss of the motor neurones. Such aminoketone 0 derivatives are preferably 2',4',6'-triemthoxy-4 (1-pyrrolidinyl) butyrophenone or pharmacologically acceptable salts and/or functional derivatives thereof The medicament to prevent, retard and ameliorate a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) and the associated loss of the motor neurones can furthermore comprise magnesium (Mg), its pharmacologically acceptable salt or its derivatives, such as magnesium oxide, magnesium aspartate, magnesium sulphate, magnesium citrate chelated magnesium, magnesium EAP in a amount sufficient to treat, to prevent, to retard, to ameliorate or to cure a motor neuron disease such as amyotrophic lateral sclerosis and the associated loss of the motor neurones.

BACKGROUND OF THE INVENTION

Present invention involves a medicinal treatment of a motor neuron disease (MND) . Motor Neuron Disease (MND) is a disease affecting the motor neurones in the brain and spinal cord. Motor neurones are the nerve cells along which the brain sends instructions, in the form of electrical impulses, to the muscles. There are two types of motor neuron, the upper motor neurones, which have long, thin nerve trunks connecting the brain to the spine. Within the spine, they connect with the lower motor neurones which, in turn, have nerve trunks connecting to the muscles of the body. Degeneration of the motor neurones leads to weakness and wasting of muscles. This generally occurs in arms or legs initially, some groups of muscles being affected more than others. Some people may develop weakness and wasting in the muscles supplying the Be:: :e:: :. :. :. :.: :: .

face and throat, causing problems with speech and difficulty chewing and swallowing. MND does generally not affect touch, taste, sight, smell or hearing, nor directly bladder, bowel, or sexual function. In the vast majority of cases, the intellect remains unaffected MND is generally a steadily progressive disease, but the rate of progression varies greatly from one person to another. s

In most cases of MND, degeneration of both the upper and lower motor neurones occurs. This condition is called Amyotrophic Lateral Sclerosis (ALS), characterized by muscle weakness, stiffness and fasciculations (muscle twitching) or, when the muscles involved in speech and swallowing are solely affected, Progressive Bulbar Palsy (PBP). There are also less common lo forms in which a more selective degeneration of either the upper motor neurones (such as Primary Lateral Sclerosis, PLS) or lower motor neurones (such as Progressive Muscular Atrophy, PMA) is observed. There is considerable overlap between these forms of MND. People with PMA in time develop upper motor neuron involvement and in both PMA and ALS some people may eventually experience speech and swallowing difficulties in varying degrees So far Motor neuron disease is dangerous an incurable disease. It causes paralysis. Motor neuron disease often begins with weakness of the muscles of the hands or feet. It eventually leads to generalised paralysis. People with motor neuron disease need help with daily activities and have a life expectancy of three to five years after their diagnosis.

ALS patients manifest symptoms associated with the loss of motor neurons, and/or the nerve cells in the spinal cord, brain stem, and motor cortex, which are normally in good control of the body's voluntary muscles. In ALS, as motor neurons die, muscles weaken and shrink, and the body manifests the early-stage symptoms of ALS. Such symptoms include, for example, unusual 2s fatigue, clumsiness, muscle weakness, slurred speech, muscle atrophy, spasticity, spinal function disorders, and convulsions. As ALS progresses, patients gradually lose the use of their hands, arms, legs, and neck muscles, ultimately becoming paralyzed. Speaking and swallowing ability are greatly compromised. Psychiatric manifestations (e. g., depression) may also result.

c. e: :e:: c:. . : :.: :: .e However, while cognitive impairment is generally not observed with ALS, some data suggest that as many as 15% of all ALS patients may experience some memory loss, behavioral changes, and problems with both judgment and simultaneously performing multiple tasks. 12 th International Symposium on ALS/MND, Oakland, Calif., Nov. 18- 20, 2001, Highlights and s Summary of Clinical Sessions, (www. alsa. org/news/newsl20301a.cfm). The usual cause of death from ALS is failure of the diaphragm muscles that control breathing. ALS patients can prolong their lives by using a ventilator, especially since bladder and bowel function, sexual function, and all five senses are unaffected. But living on a ventilator is neither desirable nor free of complications such as pneumonia (resulting from pooling of secretions or aspiration).

Evidence from early research into possible causes of ALS does not support either viral or environmental toxin theories. Also, while several autoimmune theories have been advanced in the past decade, immunosuppressive therapy, including drug (e.g., azathioprine) treatment, is plasmapheresis, or intravenous immunoglobulin injection, has been ineffective in combating ALS. Even the very potent immunosuppression method of total lymph node irradiation proved unsuccessful. Annals of Neurology, Editorial, "Amyotrophic Lateral Sclerosis: Theories and Therapies," Vol. 35, 1994, pp. 129- 130. Currently, one area of ongoing investigation is the use of growth factors, such as Insulin- like growth factor 1 (IGF- 1 or Myotrophin), ciliary neurotrophic factor (CNTF), and, most recently, vascular epithelial growth factor (VEGF) that have shown protection of motor neurons in animal models and cell culture systems. The ALS Association has announced enrollment, beginning in late winter 2002, for a clinical research trial of IGF-1. 12th International Symposium on ALS/MND, Oakland, Calif., Nov. 18-20, 2001, Highlights and Summary of Clinical Sessions, (www.alsa. org/news/ewsl20301a.cfm). The effectiveness of growth factor therapy, however, may be limited by the degradation of the protein in the liver, before it crosses the blood brain barrier (BBB).

Another avenue of therapy is in the regulation of brain glutamate levels, based on abnormally high glutamate concentrations found in cerebrospinal fluid of some ALS patients. An abundance of the glutamate transporter in astrocytes (cells surrounding the neurons) known as EAAT2, c e e I e c e e e involved in the removal of excess glutamate, is decreased in the cortex and spinal cord of patients with ALS and in mouse models. A high glutamate level leads to "excitotoxicity" (which is the activation of glutamate receptors), a flooding of neurons with calcium, and a host of damaging downstream events. Other studies have linked excitotoxicity to high zinc levels as a cause of motor neuron death. Id. From these findings, a promising area of investigation is in the use of glutamate antagonists, which inhibit the release of glutamate in the brain. Preliminary results with the FDA- approved drug riluzole (Rilutek) indicate potential for increasing the life expectancy of ALS patients. However, double blind and controlled studies are needed to confirm these observations. Id. One drawback of riluzole therapy is the need for regular monitoring for lo potentially high levels of liver enzymes, which are selected may require discontinuing riluzole drug therapy. Other glutamate antagonists of interest include dextromethorphan and lamotrigine.

As with all such ALS drugs, their potential to degrade before crossing the BBB remains a significant concern.

Finally, the identification of mutations in the gene encoding for the enzyme superoxide dismutase (SOD) in FALS has been a landmark in ALS research. SOD mutations are present in 15-20% of FALS cases. SOD converts the superoxide free radical anion (O 2-) to hydrogen peroxide (H202), which is further detoxified by enzymes such as catalase. The importance of SOD in handling free radicals during oxidative stress has been recognized for some time, leading some scientists to believe that mutant SOD may result in motor neuron destruction through the action of an excessive amount of free radicals. Studies of SOD treatment alone or in combination with other drugs such as riluzole for the treatment of both sporadic and familial ALS are ongoing.

Under certain conditions of chronic neurodegeneration, neuroinflammation may be observed.

However, the functional consequences of chronic inflammatory processes in the brain are not well understood.

An efficecnt treatment of ALS is currently non existing.

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Present invention and the subject matter herein claimed and disclosed have thus solved the above mentioned long-felt needs in the art since it demonstrates that a medicament comprising buflomedil (2',4',6'-trimethoxy4-(1-pyrrolidinyl) butyrophenone) or a pharmaceutical acceptable salts or a functional derivative thereof and a medicament further comprising magnesium or a pharmaceutically acceptable salt or functional derivatives can be used to decrease discomfort caused by a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) and moreover can be used to prevent, retard and ameliorate a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) 0 We found that a medicament comprising the aminoketone derivative, buflomedil (2',4',6'trimethoxy-4-( 1 -pyrrolidinyl) butyrophenone can ameliorate or cure ALS and that such medicament comprising the aminoketone derivative, buflomedil (2',4',6'- trimethoxy-4-( 1- pyrrolidinyl) butyrophenone) can be combined with magnesium to even treat more efficiently ALS.

The aminoketone derivative such as buflomedil or 2',4',6'-trimethoxy-4-(1pyrrolidinyl) butyrophenone, has currently been used as a vasodilator to treat vasculorpathy and diabetic angiopathy (e.g. arteriosclerosis, diabetes or endoangioitis obliterans). Magnesium on the other hand is involved in more than 300 different enzymatic reactions, including carbohydrate utilisation, ATP metabolism, muscle contraction, transmembrane ion transport, and the synthesis of fat, protein, and nucleic acids.

ILLUSTRATIVE EMBODIMENTS OF THE INVENTION

Amyotrophic lateral sclerosis (ALS) is one of several, clinically defined, motor neuron diseases (MNDs). ALS afflicts 1.5 times more men than women. In about two thirds of cases, the onset of the disease occurs between ages 50 and 70. James T. Caroscio, et al., "Amyotrophic Lateral Sclerosis: Its Natural History," Neurologic Clinics, Vol. 5, No. 1, February 1987, pp. 1-8.

Overall, ALS afflicts 5 to 10 people out of every 100,000 people. The progression of the disease is rapid. Most patients die within 5 years of onset. About 5-10% of ALS cases, known as familial ALS (FALS), are inherited. Although FALS is clinically indistinguishable from the "sporadic" f

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form of ALS, there is no predominance of FALS in men as with sporadic ALS. However, the mean age of ALS onset is comparatively earlier. J. de Belleroche, et al., "Amyotrophic Lateral Sclerosis: Recent Advances in Understanding Disease Mechanisms," J. Neuropathol. and Exp.

Neurol., Vol. 55, No. 7, July 1996, pp. 747-757. No single test can diagnose ALS. Because of the s slow onset of the disease, diagnosis of ALS is usually difficult in its early stages. By the time of positive diagnosis, the disease has generally progressed for 1-2 years.

The invention thus involves a novel medicament and treatment of a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS).

In a preferred embodiment 2', 4', 6'-trimethoxy-4-(1-pyrrolidinyl) butyrophenone or p-desmethyl- 2',4',6'-trimethoxy-4-(1-pyrrolidinyl) butyrophenone, their hydrochloride or any pharmaceutically acceptable salt or derivatives there of may administered orally in a regime of 5 to 5000 mg/patient/day, more preferably 20 to 1000 mg/patient/day, and yet more preferably 150 to 800 mg/patient/day and most preferably 300 to 600 mg/patient/day. A possible daily dose can for instance be 7 to 10 mg/kg body weight. The active compound may be delivered as a solid medicine in pill or tablet form and alternatively as liquid, semi-solid. The parenteral administration form may be an isotonic injection solution. The daily dose parenteral dose could be 3 to 6 mg/kg body weight Oral dosage forms are preferred for those therapeutic agents that are orally active, and include tablets, capsules, caplets, solutions, suspensions and/or syrups, and may also comprise a plurality of granules, beads, powders or pellets that may or may not be encapsulated. Such dosage forms are prepared using conventional methods known to those in the field of pharmaceutical 2s formulation and described in the pertinent texts, e.g., in Remington: The Science and Practice of Pharmacy, 20th Edition, Gennaro, A.R., Ed. (Lippincott, Williams and Wilkins, 2000). Tablets and capsules represent the most convenient oral dosage forms, in which case solid pharmaceutical carriers are employed.

Tablets may be manufactured using standard tablet processing procedures and equipment. One method for forming tablets is by direct compression of a powdered, crystalline or granular at:: :e::: e. ce e e composition containing the active agent(s), alone or in combination with one or more carriers, additives, or the like. As an alternative to direct compression, tablets can be prepared using wet- granulation or dry-granulation processes. Tablets may also be moulded rather than compressed, starting with a moist or otherwise tractable material; however, compression and granulation s techniques are preferred.

In addition to the active agent(s), then, tablets prepared for oral administration using the method of the invention will generally contain other materials such as binders, diluents, lubricants, disintegrants, fillers, stabilisers, surfactants, colouring agents, and the like. Binders are used to lo impart cohesive qualities to a tablet, and thus ensure that the tablet remains intact after compression. Suitable binder materials include, but are not limited to, starch (including corn starch and pregelatinised starch), gelatine, sugars (including sucrose, glucose, dextrose and lactose), polyethylene glycol, waxes, and natural and synthetic gums, e.g., acacia sodium alginate, polyvinylpyrrolidone, cellulosic polymers (including hydroxypropyl cellulose, hydroxypropyl methylcellulose, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, and the like), and Veegum. Diluents are typically necessary to increase bulk so that a practical size tablet is ultimately provided.

Suitable diluents include lactose, cellulose, kaolin, mannitol, sodium chloride, dry starch and powdered sugar. Lubricants are used to facilitate tablet manufacture; examples of suitable lubricants include, for example, magnesium stearate and stearic acid. Stearates, if present, preferably represent at no more than approximately 2 wt. % of the drug- containing core.

2s Disintegrants are used to facilitate disintegration of the tablet, and are generally starches, clays, celluloses, algins, gums or crosslinked polymers. Fillers include, for example, materials such as silicon dioxide, titanium dioxide, alumina, talc, kaolin, powdered cellulose and microcrystalline cellulose, as well as soluble materials such as mannitol, urea, sucrose, lactose, dextrose, sodium chloride and sorbitol. Stabilisers are used to inhibit or retard drug decomposition reactions that include, by way of example, oxidative reactions. Surfactants may be anionic, cationic, amphoteric or non-ionic surface active agents.

I: Id: c : The dosage form may also be a capsule, in which case the active agent-containing composition may be encapsulated in the form of a liquid or solid (including particulates such as granules, beads, powders or pellets). Suitable capsules may be either hard or soft, and are generally made s of gelatine, starch, or a cellulosic material, with gelatin capsules preferred. Two-piece hard gelatine capsules are preferably sealed, such as with gelatine bands or the like. See, for example, Remington: The Science and Practice of Pharmacy, which describes materials and methods for preparing encapsulated pharmaceuticals. If the active agent-containing composition is present within the capsule in liquid form, a liquid carrier is necessary to dissolve the active agent(s) . The lo carrier must be compatible with the capsule material and all components of the pharmaceutical composition, and must be suitable for ingestion.

Solid dosage forms, whether tablets, capsules, caplets, or particulates, may, if desired, be coated so as to provide for delayed release. Dosage forms with delayed release coatings may be Is manufactured using standard coating procedures and equipment. Such procedures are known to those skilled in the art and described in the pertinent texts, e.g., in Remington, supra. Generally, after preparation of the solid dosage form, a delayed release coating composition is applied using a coating pan, an airless spray technique, fluidised bed coating equipment, or the like. Delayed release coating compositions comprise a polymeric material, e.g., cellulose butyrate phthalate, cellulose hydrogen phthalate, cellulose proprionate phthalate, polyvinyl acetate phthalate, cellulose acetate phthalate, cellulose acetate trimellitate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate, dioxypropyl methylcellulose succinate, carboxymethyl ethylcellulose, hydroxypropyl methylcellulose acetate succinate, polymers and copolymers formed from acrylic acid, methacrylic acid, and/or esters thereof. 2s

Sustained release dosage forms provide for drug release over an extended time period, and may or may not be delayed release. Generally, as will be appreciated by those of ordinary skill in the art, sustained release dosage forms are formulated by dispersing a drug within a matrix of a gradually bioerodible (hydrolysable) material such as an, insoluble plastic, a hydrophilic polymer, or a fatty compound, or by coating a solid, drug containing dosage form with such a material. Insoluble plastic matrices may be comprised of, for example, polyvinyl chloride or t. rye; : c:. :e: :: :: ees polyethylene. Hydrophilic polymers useful for providing a sustained release coating or matrix cellulosic polymers include, without limitation: cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, ethyl cellulose, cellulose acetate, cellulose acetate phthalate, cellulose acetate trimellitate, hydroxypropylmethyl cellulose phthalate, hydroxypropylcellulose phthalate, cellulose hexahydrophthalate, cellulose acetate hexahydrophthalate, and carboxymethylcellulose sodium; acrylic acid polymers and copolymers, preferably formed from acrylic acid, methacrylic acid, acrylic acid alkyl esters, methacrylic acid alkyl esters, and the like, e.g. copolymers of acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylate and/or ethyl lo methacrylate, with a terpolymer of ethyl acrylate, methyl methacrylate and trimethylammonioethyl methacrylate chloride (sold under the tradename Eudragit RS) preferred; vinyl polymers and copolymers such as polyvinyl pyrrolidone, polyvinyl acetate, polyvinylacetate phthalate, vinylacetate crotonic acid copolymer, and ethylenevinyl acetate copolymers; zein; and shellac, ammoniated shellac, shellac-acetyl alcohol, and shellac n-butyl stearate. Fatty compounds for use as a sustained release matrix material include, but are not limited to, waxes generally (e.g., camauba wax) and glyceryl tristearate.

Alternatively transepidermal effective amounts of 2',4',6'-trimethoxy-4(1-pyrrolidinyl) butyrophenone or of p-desmethyl-2',4',6'-trimethoxy-4-(1pyrrolidinyl) butyrophenone, their hydrochloride or any pharmaceutically acceptable salt or derivatives thereof may be administered topically on the respective areas of motor neuron loss. The transdermal administration of 2',4',6'- trimethoxy-4-( 1 -pyrrolidinyl) butyrophenone or of p-desmethyl-2',4',6'- trimethoxy-4-( 1- pyrrolidinyl) butyrophenone, their hydrochloride or any pharmaceutically acceptable salt or derivatives thereof can be transdermal electromotive administration, the transdermal absorption being accelerated by use of an electrode-drug receptacle attached to the patients. For such topical treatment the pharmaceutical product can be used as liquid, semi-solid or solid medicine. Liquid medicines are solutions, suspensions, emulsions or dispersions of the above-cited active ingredients or combinations of active ingredients as drops, tinctures and sprays. As semi-solid medicines, for example, gels, ointments, creams and foams are used while, for example, powders, toilet powders, granulates, pellets and microcapsules are used as solid medicines.

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If the pharmaceutical product containing as active ingredient 2',4',6'trimethoxy-4-(1 pyrrolidinyl) butyrophenone, its hydrochloride or any pharmaceutically acceptable salt or derivatives thereof, is used as a liquid, it is recommended to use as far as possible irritation-free diluting agents, as for example water, monovalent alcohols, especially ethanol, polyvalent alcohols, especially glycerine and/or propanediol, polyglycols, especially polyethylene glycols andlor miglyols, glycerine formal, dimethylisosorbide, natural and synthetic oils and/or esters.

For the production of semi-solid products, as for example gels, ointments, creams and foams, in addition to the above-cited diluting agents basic materials, as for example bentonite, veegum, lo guar flour and/or cellulose derivatives, especially methylcellulose and/or caboxymethylcellulose, are suitable. The buflomedil hydrochloride, maleate and/or alkali and/or alkaline earth salts may be in the form of a physico-chemical complex with a phospholipid selected from the group consisting of lecithin, cephalin, phosphatidylserine, phosphoinositide, and phosphatidic acid, or mixtures thereof in the form of a cream, an ointment, a pomade, a gel, or an emulsion to the area is to be treated. The process of manufacture of such complexes has been described by Bertini Curri in US5,280,020.

Furthermore, instead of the above-cited basic materials or in addition to these materials polymers of vinylalcohol and vinylpyrrolidone, alginates, pectines, polyacrylates, solid and/or liquid polyethylenglycols, paraffins, fatty alcohols, vaseline and/or waxes, fatty acids and/or fatty acid esters are used. It is possible to use the above-cited active ingredients without filler for the production of solid products, as for example powders, toilet powder, granulates, pellets and microcapsules. The pharmaceutical product described here is especially suited for the attention of such of the above-described diseases which are in a very progressed stage so that at first an 2s increased concentration of active ingredients is necessary. With less serious disease conditions or with progressive healing of the disease such embodiments of the solid pharmaceutical product are used which contain fillers, as for example colloidal silicic acid, powdered soapstone, milk sugar, starch powder, sugar, cellulose derivatives, gelatin, metal oxides and/or metal salts, wherein the concentration of the active ingredient or of the combination of active ingredients varies between 0.001% by weight and 50% by weight.

i. . .' . 1 ' A suitable kind of pharmaceutical form may be a topical deliver form of the above-described active ingredient, which is made by the application of the solid, liquid or semi-solid pharmaceutical product onto a gauze strip, a compress or a plaster so that such a gauze strip, such a compress or such a plaster then is only locally applied onto the spot which is to be treated. The s pharmaceutical product can be filled into the known receptacles, as for example bottles, tubes, toilet powder boxes and baby powder boxes as well as seal edge bags, which are possibly provided with metering means, as for example droplet forming means, metering valves or metering chambers.

lO Buflomedil hydrochloride is synonym of 4-(l-Pyrrolidinyl)-1-(2,4,6trimethoxyphenyl)-1- butanone has CAS Number 55837-25-7 and molecular Formula: C17H25NO4 HCI and is describable by the structure formula as displayed in Fig 1.

For the manufacture of the pharmaceutical product of present examples the 2', 4',6'-trimethoxy-4- s (l-pyrrolidinyl) butyrophenone indicated above was used. Instead of the 2',4',6'-trimethoxy-4-(1- pyrrolidinyl) butyrophenone or in addition to it salts, especially the corresponding hydrochloride, maleate and/or alkali and/or alkaline earth salts of the 2',4',6'-trimethoxy-4-(1-pyrrolidinyl) butyrophenone, which can be sulfated and/or sulfonated on the aromatic moiety and/or the pyrrolidine moiety, can be used for the manufacture of the pharmaceutical product. The pharmaceutical products which can be used for the treatment of a motor neuron disease such as ALS or the associated motor neuron loss, in addition to the above-cited active ingredients or instead of the above-cited ingredients, can also contain derivatives, preferably pharmacologically active metabolic products (metabolites), of the 2',4', 6'-trimethoxy-4-( l -pyrrolidinyl) butyrophenone, especially the pdesmethyl-2',4',6'-trimethoxy-4-( l -pyrrolidinyl) butyrophenone.

The pharmaceutical compound buflomedil may be administered as the hydrochloride, maleate and/or alkali and/or alkaline earth salts. It may be bound to a cation-exchange resin for instance a buflomedil resinate complex. Buflomedil hydrochloride can for instance be bound to cationexchange resins by soaking the resin in an aqueous solution of the drug, which renders it particularly suitable for oral administration. Process for making a taste-masked buflomedil c c #.

C resinate are described by R. A. Honeysett et al. "Taste- masked Buflomedil preparation" in EP0501763.

2', 4', 6'-triemthoxy-4 (1-pyrrolidinyl) butyrophenone, to which the examples of present invention relates is marketed as Buflomedil hydrochloride. It is a peripheral vasodilator which has been sold under the trade names BUFENE (I.C.I.), LOFTIL (ABBOT), BUFLAN (PIERREL), IRRODAN (BIOMEDICA FOSCANA) and FLOMED (PULITZER). Amongst the other aminoketones which are classified as peripheral vasodilators and which are more widely used in the treatment of functional and organic peripheral vasculopathy are for instance aminoketone derivatives of 0 fluoroglucinol such as 4-(pyrrolidinyl)-1 (2,4,6-trimethoxyphenyl)-1- butanone; 2', 4', 6' trimethoxy-4-(1'-pyrrolidinyl) butyrophenone; (2,4,6- trimethoxyphenyl) (3-pyrrolidinopropyl) ketone, (2,4,6-Trimethoxyphenyl) (3-piperidinopropyl) ketone; (2,4,6-triethoxyphenyl) (3 diethylaminopropyl) ketone; (2,4,6-trimethoxyphenyl) [4-J3-hydroxyethylpiperazino) methyl] ketone dihydrochloride and (2,4,6-triethoxyphenyl) (3pyrrolidinopropyl) ketone of which s preferably their salts thereof with pharmaceutically acceptable acids can be used to treat a motor neuron disease such as ALS or the associated motor neuron loss or to manufacture a medicament to treat a motor neuron disease such as ALS or the associated motor neuron loss. The synthesis of these structures have been described in German Pat. 2,122,144 (1971), in U.S. Pat. No. 3,895,030 (1971) and in Italian patent No. 1,120,968. And their use can be within the scope of present invention. These compounds are known to act as vasodilator agent with calcium antagonistic and alpha-adrenoceptor action.

Buflomedil, for instance, is a competitive inhibitor of alpha-adrenergic receptors and acts as a non-specific calcium antagonist. As a vasodilator,it acts via mechanisms of alpha-adrenoceptor blockade and calciummobilisation-blocking. Buflomedil is a competitive inhibitor of alpha adrenergic receptors, not selective for either the alpha- 1- or alpha-2adrenergic receptor subtype.

Addition of buflomedil can totally block the vasoconstriction elicited by phenylephrine.

Buflomedil hydrochloride improves perfusion in the impaired microcirculation of the peripheral and central vascular beds. The vascular activity of buflomedil is related to the two pharmacological components: an alpha- I effect and alpha-2 adrenolytic and to a direct action on e he c C the microcirculatory structures related to an effect on the calcium uptake at the membrane level of the perivacular myocytes. By its adrenolytic non-specific alpha action, buflomedil is opposed locally to the vasoconstriction effect adrenaline in for instance response to stress or to cold. This action is found primarily in these arteries which have abundant alpha-receptors such as the muscular peripheral arteries of the distribution system. By its calcium-channel-blocking action properties buflomedil opens the contracted pre-capillary sphincters and thus restores a functional microcirculation.

An embodiment of present invention is a method of treating of a motor neuron disease such as lo amyotrophic lateral sclerosis and the associated degeneration of motor neurones, which comprises treating an affected patient with a therapeutically effective amount of a pharmaceutical composition containing as active ingredient a therapeutically effective amount of alpha adrenoceptor antagonist (competitive inhibitor of a alpha-adrenergic receptors) and a calcium antagonist (calcium-channel-blocker) or a compound which comprises both properties.

Calcium calcium blockers are widely used as vasodilator drug. Their mechanism by which they act as vasodilator is based on a calciumantagonistic effect whereby they interfere with the transmembrane transfer of calcium in the smooth arterial muscles or act on platelet sludging and on erythrocyte deformability. Despite the various chemical structures of calcium channel blockers, they have one primary mode of action. The cells e.g. cardiac or myocytes release calcium ions through subcellular structures and thereby activate the contractile proteins. A calcium selective channel allows the influx of calcium ions from the extracellular space into the cell, which also triggers the release of calcium from the cell structure. The calcium channel blockers block the calcium selective channel, and hence act primarily as a vasodilator. Potent calcium channel blockers, other than the aminoketone derivates, with marked vasodilator action are 3,5-pyridinedicarboxylic acid, 1,4dihydro-2,6-dimethyl-4-(2-nitrophenyl)- dimethyl ester; 1,4-Dihydro-2,6-dimethyl-4-(3-nitrophenyl) methyl 2-(methyl (phenylmethyl) amino) -3,5 pyridinecarboxylic acid ethyl ester, brovincamine (Tanaka, Y. , Arzneimittelforschung, 44, 803 8), nifedipine, testosterone (K.M. English et al. J. Endocrinol. Invest. 2002, 25: 455-458), Felodipine. Cohn, J. N. et al. Circulation 96: 856- 863). Various others calcium channel blockers used or under investigation to widen the blood vessels are felodipine, amlodipine, nifedipine, . e C ee. e ece C C . . . verapamil HCI, nicardipine HCI, diltiazem HCI, aranidipine, atosiban, barnidipine, cilnidipine, docosahexaenoic acid, efonidipine HCL, fasudil, isradipine, lacidipine, lercanidipine, lomerizine, manidipine, nifelan, nilvadipine, nimodipine, teczem, verelan, plendil, nisoldipine, bepridil HCI, NS-7, NW- 1015, SB-237376, SL-34. 0829-08, terodiline, R-verapamil, bisaramil, CAI, ipenoxazone, JTV-519, S- 312d, SD-3212, tamolarizine, TA-993, vintoperol, YM-430, CHF 1521, elgodipine, nitrendipine, furnidipine, L-651582, oxodipine, ranolazine, AE-0047, azelnidipine, dotarizine, lemildipine, pranidipine, semotiadil, temiverine HCI, tenosal, vatanidipine HCI, and ziconotide. They can without limitation be used within the scope of this invention.

The adrenoceptor family was first divided into two subtypes, the a- and badrenoceptors, as determined by pharmacological studies in isolated tissue (Ahlquist, 1948). A quarter of a century later, the a- adrenoceptors were further subdivided based on their anatomical location, with the a adrenoceptors located on peripheral sympathetic nerve terminals designated a2 adrenoceptors, and those located post- synaptically designated al-adrenoceptors (Langer, 1974). This anatomical classification rapidly gave way to the identification of pharmacological differences between the a-adrenoceptors, notably the ability of yohimbine and rauwolscine to act as a2-adrenoceptor antagonists. Subsequent studies using pharmacological and molecular biology techniques have further subdivided the a adrenoceptor family; three subtypes within each group have now been cloned and pharmacologically characterised. The al- adrenoceptor subtypes have been classified as the ala-, alB- and alD- adrenoceptors and the a2-adrenoceptors have been classified as the a2A- (a2D- species variation of the human a2A-), a2B- and a2C-adrenoceptors.

Alpha-adrenoceptor antagonist or adrenolytic antogonists with a nonspecific alpha action, which are currently used as a pharmaceutical are for instance: cetiedil (of Innothera), dihydroergocristine (Poli), dihydroergotamine (bioMerieux-Pierre Fabre), dihydroxyacetone (Vines), etoperidone, Angelini (Wyeth), labetalol (GlaxoSmithKline), trimazosin (Pfizer), ciclonicate (Poli), dapiprazole (Angelini), amosulalol (Yamanouchi), arotinolol (Sumitomo), naftopidil (Hoffmann-La Roche), terazosin (Abbott), urapidil (Altana), phentolamine (Zonagen), phentolamine (Novartis), dihydroergotamine SR (Ethypharm), Alphadihydroergocryptine (Poli).

Nonselective alpha-adrenergic antagonists currently underdevelopment are for instance e ë e e ee e e e e e e e ce e e e e c e ace e e e e see e e e e e e e eee e e e e Naftidrofuryl SR (Ethypharm), dihydroergotamine (POZEN), AGN- 195795 (ACADIA), ergotamine (Sheffield), dihydroergotamine, (Sheffield). They can without limitation be used within the scope of this invention.

s Present invention thus involves a method of treating, preventing or ameliorating a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) in a subject comprising inhibiting the alpha receptors in the arteries and calcium-channel-blocking of the myocytes at the neruones.

Furthermore the present invention involves a method of treating, preventing or ameliorating a lo motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) in a subject comprising administering to said subject an effective amount of a composition comprising a calcium channel blockers and an alpha-adrenoceptor antagonist (adrenolytic antagonists) wherein calcium channel blockers and alpha-adrenoceptor antagonist is a aminoketone derivative of fluoroglucinol or the hydrochloride form or any pharmaceutical acceptable salts thereof The invention also involves a method of treating, preventing or ameliorating a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) in a subject comprising administering to said subject an effective amount of a composition comprising a calcium channel blockers and an alphaadrenoceptor antagonist (adrenolytic antagonists) wherein calcium channel blockers and alpha-adrenoceptor antagonist is an aminoketone derivatives of fluoroglucinol selected from the group consisting of 4-(pyrrolidinyl)-1 (2,4,6-trimethoxyphenyl) -1- butanone, 2', 4', 6'- trimethoxy-4-( 1'-pyrrolidinyl) butyrophenone; (2,4,6-trimethoxyphenyl) (3-pyrrolidinopropyl) ketone, (2,4,6-Trimethoxyphenyl) (3-piperidinopropyl) ketone, (2,4,6- triethoxyphenyl) (3diethylaminopropyl) ketone, (2,4,6-trimethoxyphenyl) [4-13-hydroxyethylpiperazino) methyl] Is ketone dihydrochloride and (2,4,6-triethoxyphenyl) (3-pyrrolidinopropyl) ketone of which preferably their salts thereof Yet an other embodiment of present invention is a method of treating, preventing or ameliorating a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) in a subject comprising administering to said subject an effective amount of a composition comprising a calcium channel blockers and an alpha-adrenoceptor antagonist (adrenolytic antagonists) wherein . e e . . . . calcium antagonist (calcium blocker) and alpha-adrenoceptor antagonist is an aminoketone derivatives of fluoroglucinol of the group consisting of 4-(pyrrolidinyl)- 1 (2,4,6 trimethoxyphenyl)- 1 -butanone, 2',4',6'-trimethoxy-4-(1'-pyrrolidinyl) butyrophenone and (2,4,6 trimethoxyphenyl) (3-pyrrolidinopropyl) ketone. s

The invention involves furthermore a method of treating, preventing or a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) in a subject comprising administering to said subject an effective amount of a composition comprising a calcium channel blockers and an alpha-adrenoceptor antagonist (adrenolytic antagonists) wherein calcium channel blockers and o alpha-adrenoceptor antagonist is 2',4',6'-trimethoxy-4-(1-pyrrolidinyl) butyrophenone or p desmethyl-2',4',6'-trimethoxy-4-(1 -pyrrolidinyl) butyrophenone or the addition salts thereof The invention involves furthermore a method of treating, preventing or ameliorating a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) in a subject comprising: (a) administration to said subject an effective amount of a calcium channel blockers and (b) co administration to said subject an effective amount of alpha-adrenoceptor antagonist (adrenolytic antagonists) wherein said calcium channel blockers is selected of the group consisting of felodipine, amlodipine, nifedipine, verapamil HCI, nicardipine HCI, diltiazem HCI, aranidipine, atosiban, barnidipine, cilnidipine, docosahexaenoic acid, efonidipine HCL, fasudil, isradipine, lacidipine, lercanidipine, lomerizine, manidipine, nifelan, nilvadipine, nimodipine, teczem, verelan, plendil, nisoldipine, bepridil HCI, NS-7, NW- 1015, SB-237376, SL-34.0829-08, terodiline, R-verapamil, bisaramil, CAI, ipenoxazone, JTV-519, S-312d, SD-3212, tamolarizine, TA-993, vintoperol, YM-430, CHF- 1521, elgodipine, nitrendipine, furnidipine, L-651582, oxodipine, ranolazine, AE-0047, azelnidipine, dotarizine, lemildipine, pranidipine, semotiadil, temiverine HCI, tenosal, vatanidipine HCI, and ziconotide and said alpha-adrenoceptor antagonist is selected from the group consisting of cetiedil, dihydroergocristine, dihydroergotamine, dihydroxyacetone, etoperidone, Angelini, labetalol, trimazosin, ciclonicate, dapiprazole, amosulalol, arotinolol, naftopidil, terazosin, urapidil, phentolamine, phentolamine, dihydroergotamine SR, Alphadihydroergocryptine, Naftidrofuryl SR, dihydroergotamine, AGN 195795, ergotamine, dihydroergotamine.

* e e c * * # * * * * * * The invention may also concern to the use of the compounds and the combination of said compounds with adrenolytic antagonists and calcium channel blocking properties, mentioned above, to manufacture a medicament for treating, preventing or ameliorating a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS).

Magnesium which in an example of present invention has been combined buflomedil and had been found to enhance the cure of a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS). Magnesium is the fourth most abundant cation in the body, following calcium, sodium, and potassium, and the second most abundant intracellular cation (after potassium). It is lo involved in more than 300 different enzymatic reactions, including carbohydrate utilisation, ATP metabolism, muscle contraction, transmembrane ion transport, and the synthesis of fat, protein, and nucleic acids. Magnesium is thus a physiologic necessity, but can also used as a pharmacological compound. Magnesium is, for instance also considered to be a natural calcium channel blocker and has vasodilator properties and is an inhibitor of vascular smooth muscle cell contraction (Shechter M., et al. Circulation. 2000 Nov 7;102(19):2353-8).

An embodiment of present invention is method of treating a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) and the associated motor neuron degeneration such as spinal motor neuron degeneration, which comprises treating an affected patient with a therapeutically effective amount of a pharmaceutical composition containing as active ingredient a therapeutically effective amount of aminoketones derivatives of fluoroglucinol and preferably a aminoketones derivatives selected from the group consisting of 4-(pyrrolidinyl)- 1 (2, 4,6 trimethoxyphenyl) -l-butanone; 2', 4', 6'-trimethoxy-4- (1'pyrrolidinyl) butyrophenone; (2,4,6 trimethoxyphenyl) (3pyrrolidinopropyl) ketone, (2,4,6-Trimethoxyphenyl) (3-piperidinopropyl) ketone; (2,4,6-triethoxyphenyl) (3-diethylaminopropyl) ketone; (2,4,6trimethoxyphenyl) [4-13 hydroxyethyl-piperazino)methyl] ketone dihydrochloride and (2,4,6-triethoxyphenyl) (3 pyrrolidinopropyl) ketone or their pharmaceutically acceptable salts and most preferably 2', 4', 6' trimethoxy-4-( 1 -pyrrolidinyl) butyrophenone or p-desmethyl-2',4',6'trimethoxy-4-( 1 pyrrolidinyl) butyrophenone, in the form of their hydrochloride or any pharmaceutically acceptable salt of the active ingredient. The active ingredient may be orally or parenterally administered.

te e ce e C ce. e e Pharmaceutical compositions according to the invention can be periodically administered to a mammalian patient (e.g., a human patient), in need of such treatment, to promote motor neuron regeneration and functional recovery and thereby to treat damage to motor neurones caused by a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS). The compounds of s present invention are thus useful to enhance regeneration of the motor neurones. The present invention thus involves a method of promoting motor neuron regeneration in a mammal having a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS), the method comprising: administering to the mammal a pharmaceutical composition comprising a motor neuron regeneration stimulating amount of the compounds of present invention. The compounds lo of present invention can also be used to enhance motor recovery following a motor neuron damage by a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS).

To further enhance motor neuron repair the compounds of present invention may be combined with known neurotrophic factors, which have been shown to play an essential throphic role in the development, maintenance and regulation of neuronal function such as for instance ciliary neurotrophic factor (CNTF), glial growth factors (glial mitogenic factors), schwann cell mitogenic factors, the nerve growth factors (NGF) and other members of the NGF family, brain- derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) or platelet- derived growth factor (PDGF).

To further enhance recovery of a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) the compounds of present invention may be combined with VEGF homologous, which have been shown to play a role in mediated survival signals to motor neurons or motor neuron protection. The compounds of present invention may for instance be combined with for instance VEGFA, VEGFB or PLGF To further enhance recovery of a motor neuron disease (MND) such as amyotrophic lateral sclerosis (ALS) the compounds of present invention may be combined with known neuroprotective compounds such as the compounds of the group consisting of free radical scavenger, alpha-phenyl-tert-butyl nitrone (PBN); non-competitive antagonists of glutamate such as for instance 1-(1(2-thienyl)cyclohexyl)piperidine (TCP) or topiramate, peroxide dismutases, e # adenosine agonists such as cyclohexyladenosine (CHA), Gammaamino butyric acid (GABA) and GABA-mimetic drugs, non-NMDA receptor antagonist such as 2,3- dihydroxy-6-nitro-7- sulfamoyl-benzo(F)quinoxaline (NBQX), the green tea polyphenol (-)- epigallocatechin-3-gallate, NAALADase inhibitors such 2-(phosphonomethyl)- pentanedioic acid (2-PMPA), small s molecule inhibitors of cyclin- dependent kineses (CDKs), cyclooxygenase 2 (COX-2) inhibitors alone or in combination with creative, (IR)-1-benzo [b] thiophen-5-yl-2-[2- (diethylamino) ethoxy] ethan-l-ol hydrochloride (T-588), riluzole, an inhibitor of glutamate transmission and drug used in in amyotrophic lateral sclerosis (ALS), Cardiotrophin-l (CT- 1), nocycline, cyclin- dependent kinase CDK selective inhibitors (CDKIs) and phosphodiesterase (PDE) inhibitors such o as the selective PDE5 inhibitors (dipyridamole, T- 1032, and zaprinast) as well as the nonselective PDE inhibitor (aminophylline).

Examples

t5 The efficacy of the invention can be demonstrated on animal models for studying amyotrophic lateral sclerosis, such as the Wobbler mouse (Leestma J. E., Am. J. Pathol., 100, 821-824), a mouse model of spontaneous MND affected with progressive motor neuron degenerative disease.

Buflomedil has a potent survival-promoting effects on motor neurons in the natural model of motor neuron disease and can attenuate progressive disease in wobbler mice. Buflomedil retardes denervation muscle atrophy and reduces degeneration of the spinal motor neurons. This can be observed by prevented deterioration in paw position and walking pattern abnormalities. Grip strength and running speed are observed to be greater with buflomedil treatment than with vehicle treatment.

2s To demonstrate the invention clinical diagnosis mouse at age 3 to 4 weeks, are randomly assigned to receive Buflomedil HCI (SOmg/ml (Lofty!)) IP injected at 5 mg/kg/day or vehicle daily. 6 times/week for 4 weeks after ë e

Claims (13)

1. -

   MOTOR NEURON LOSS

   What is claimed is: 1. Use of a aminoketone derivative of fluoroglucinol or the hydrochloride form or any 0 pharmaceutical acceptable salts thereof for the preparation of a pharmaceutical composition to treat a motor neuron disease (MND).
2. 2. Use of the aminoketone derivatives according to claim 1, or any pharmaceutical acceptable salts thereof for the preparation of a pharmaceutical composition to treat MND and the associated motor neuron loss
3. 3. Use of the aminoketone derivatives according to claim 1, wherein the MND is Amyotrophic Materal Sclerosis (ALS).
4. 4. Use of the aminoketone derivatives according to claim 1, wherein the MND is Primary Lateral Sclerosis, (PLS).
5. 5. Use of the aminoketone derivatives according to claim 1, wherein the MND is Progressive Muscular Atrophy (PMA).
6. 6. Use of the aminoketone derivatives according to claim 1, for the preparation of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.
7. 7. Use of the aminoketone derivatives of claim I with magnesium, its pharmaceutically acceptable salt or its derivatives for the preparation of a pharmaceutical composition to treat or ameliorate damage to motor neurons caused by MND.
8. 8. The use of claim 7, wherein the magnesium salt or derivative is a compound selected from the group consisting of magnesium oxide, magnesium aspartate, magnesium sulphate, magnesium citrate, chelated magnesium and magnesium EAP.
9. 9. The use of any of the claims 1 to 8, wherein the aminoketone derivatives of fluoroglucinol is a compound selected from the group consisting of 4-(pyrrolidinyl)- 1 (2,4,6 trimethoxyphenyl) -1 -butanone, 2', 4', 6'-trimethoxy-4-( 1'-pyrrolidinyl) butyrophenone; r e' .

   C C

   (2,4,6-trimethoxyphenyl) (3-pyrrolidinopropyl) ketone, (2,4,6Trimethoxyphenyl) (3 piperidinopropyl) ketone, (2,476-triethoxyphenyl) (3diethylaminopropyl) ketone, (2,4,6 trimethoxyphenyl) [4-13-hydroxyethylpiperazino) methyl] ketone dihydrochloride and (2,4,6 triethoxyphenyl) (3pyrrolidinopropyl) ketone of which preferably their salts thereof
10. 10. The use of any of the claims l to 8, wherein the aminoketone derivatives of fluoroglucinol is a compound selected from the group consisting of 4-(pyrrolidinyl)-1 (2,4,6 trimethoxyphenyl)-l-butanone, 2', 4',6'-trimethoxy-4-(1'-pyrrolidinyl) butyrophenone and (2,4,6-trimethoxyphenyl) (3-pyrrolidinopropyl) ketone.
11. 11. The use of any of the claims 1 to 8, wherein the aminoketone derivatives of fluoroglucinol is to 2',4',6'-trimethoxy-4-(1-pyrrolidinyl) butyrophenone or p-desmethyl-2',4',6'-trimethoxy-4-(1 pyrrolidinyl) butyrophenone or the addition salts thereof
12. 12. The use of 2',4',6'-trimethoxy-4-(1-pyrrolidinyl) butyrophenone or pdesmethyl-2',4',6' trimethoxy-4-(1-pyrrolidinyl) butyrophenone of claim 11, bound to a cation-exchange resin for the preparation of a pharmaceutical composition for the treatment of damage to motor neurones caused by MND.
13. 13. A method of treating or ameliorating damage to the motor neurones caused by MND in a subject comprising administering to said subject an effective amount of a composition comprising a aminoketone derivatives of fluoroglucinol of any of the claims 1 to 12 or an hydrochloride form or any pharmaceutical acceptable salts thereof. 2l