HU226376B1 - Pharmaceutical compositions containing nitrogen containing heterocyclic compounds for treating inflammatory disorders of the respiratory tract. - Google Patents

Abstract

N-containing heterocylic cpds, as AMPA receptor, modulator cpds. (as benzo-diazepines, and quinoxaline derivs.) are used in treatment of mammalian respiratory tract inflammation and are esp. useful in treating asthma and asthma related diseases.

Description

BACKGROUND OF THE INVENTION

FIELD OF THE INVENTION

The present invention relates to the use of nitrogen-containing heterocyclic compounds for the manufacture of a medicament for the prevention, treatment or amelioration of acute and chronic inflammatory diseases of the mammal, including allergic inflammations such as asthma and asthma-related diseases.

Summary of related literature

Inflammation is a multi-step cascade process, any part of which may be the subject of potential therapeutic intervention. In short, inflammation involves the infiltration of immunologically competent cells (e.g., eosinophils, mast cells, activated T-lymphocytes) at the site of injury, where they, together with resident cells, release bioactive mediators (e.g., histamine, proteases, cytokines, and chemokines). , which increase permeability of adjacent blood vessels, attract and stimulate waiting cells. The altered permeability of blood vessels results in fluid excretion at the site of injury, followed by further influx of reactive leukocytes and their final outflow to the injured area. (For review, see Trowbridge and Emling, Inflammation: A Review of the Process Quintessence Pub. Co., 1997). Collagen and mucus secreted by resident cells (smooth muscle and epithelial cells or connective tissue cells stimulated by released mediators) and the proliferation of these cells trigger pathological changes (such as airway obstruction) and contribute to their development.

Inflammation is associated with a number of lung diseases, such as non-allergenic and allergen-induced bronchial asthma, any inflammatory lung disease, acute or chronic bronchitis, chronic bronchitis-related secondary inflammatory reactions, may play a role, including, but not limited to, idiopathic pulmonary fibrosis and any other autoimmune lung disease. Asthma is probably one of the most common forms of inflammatory disease of the lungs, affecting the large and small lungs of the lungs. It affects 5-10% of the human population, estimated to result in 27 million medical visits, 6 million lost working days, and 90.5 million days of restricted activity per year. Induced asthma disease and mortality in women worldwide [Plaut and Zimmerman, "Allergy and Mechanisms of Hypersensitivity" in Fundamental Immunology, 3 ^rd Ed., Paul (ed.), Raven Press, New York, NY, at 1399 (1993)] .

Conventional anti-asthma treatments have so far been based on strict avoidance of allergy-causing substances, which are fundamentally difficult to achieve, and have been based on therapeutic methods that employ pharmaceuticals with adverse side effects and sub-optimal pharmacokinetic properties. Β ₂ -adrenergic agonists used to treat bronchospasm have no effect on inflammatory airways or bronchial hyperreactivity [Palmer et al., New Engl. J. Med. 331: 1314 (1994)]. In addition, regular or prolonged use of β ₂ -adrenergic agonists has little effect on asthma, increases airway sensitivity to allergens, and provides little protection against bronchial contractility (Bhagate et al., Chest 108: 1235 (1995)). In addition, chronic administration of β ₂ -adrenergic preparations alone, by inducing reduced regulation of β ₂ -adrenergic receptors, is likely to exacerbate bronchial hyperreactivity. Theophyllin (an anti-asthma methylxanthine) exhibits markedly variable absorption and elimination characteristics. While corticosteroids are relatively safe in adult patients but toxic in children, they cause adrenal degeneration and reduced bone density and growth [Woolock et al., Am. Respir. Crit. Care Med. 153: 1481 (1996)]. Cromolyn, which is used to prevent asthmatic episodes, is only effective in preventing an asthmatic reaction when administered prior to a seizure [Volcheck et al., Postgrad. Med. 104 (3): 127 (1998)]. Antihistamines sometimes prevent or stop allergic asthmatic episodes, especially in children, but are often only partially effective because histamines are only one class of mediators associated with many inflammations. [Cuss, Asthma as an Inflammatory Disease , O'Byrne, Ed., Dekker, Inc., New York, 199 (1990) and O'Byrne, "Airway Inflammation and Asthma, in Asthma as an Inflammatory Disease,"O'Byrne, Ed., Dekker, Inc. , New York, NY, 143 (1990)].

So, current drug modifications have a number of disadvantages. Conventional agents generally have a relatively short duration of action and may be partially or completely ineffective when administered after the antigen reaction has occurred. In addition, since the use of agents such as? ₂ -adrenergic agonists and corticosteroids has serious side effects, the therapeutic margin for these types of agents is relatively narrow and patients using them should be closely monitored (see, e.g., WO 94/06783 and WO 94/06783). 99/06025; and U.S. Patent Nos. 5,690,910 and 5,980,865). Another clinical study conducted with inhaled corticosteroids showed only a temporary improvement in respiratory function in asthmatic children aged 5-11 years after the first year of treatment and a decrease in placebo over the next 3 years [The Childhood Asthma Management Program Research Group, N. Engl. J. Med., 343: 1054 (2000)]. This observation can best be explained by the development of abnormal airways (a characteristic feature of asthma) that is difficult to detect with corticosteroids.

HU 226 376 zen1 can be cured [Davies, Curr. Opin. Allergy Clin. Immunol., 1:67 (2001)].

Recently, glutamate receptors have been found in cells outside the central nervous system (Skerry and Genever, Trends in Pharmacol. Sci., 22: 174 (2001)] and NMDA receptors in the lungs have been identified (Said et al., Proc. Natl. Acad. Sci. (USA) 93: 4688 (1996)]. Here, activation of the NMDA receptor results in an acute edema lung injury, similar to that observed in "acute respiratory distress syndrome" [Said et al., Proc. Natl. Acad. Sci. USA 93: 4688 (1996)]. It has also been suggested that inflammatory injury may result from activation of NMDA receptors in the asthmatic airways [Said, Trends in Pharmacol. Sci. 20: 132 (1999)]. Glutamate antagonists also showed activity outside the nervous system. Rzeski et al., Proc. Natl. Acad. Sci. USA, 98: 6372 (2001)] demonstrated in vitro antiproliferative activity of both AMPA and NMDA antagonists on a panel of tumor cell types.

The identification of moieties capable of inhibiting AMPA receptor activation, thus facilitating the identification of both competitive and non-competitive AMPA receptor antagonists, has been of great interest. The utility of AMPA receptor modulators as potential therapeutic agents for the treatment of a number of central nervous system disorders, such as ischemia, epilepsy, schizophrenia, memory impairment and drug dependence, is currently being investigated [Id. for example, Lees, Drugs, 59:33 (2000) as a comprehensive publication].

AMPA receptor antagonists are reported in numerous patents and patent applications, including the following U.S. Patents 4,812,458; 5,192,792; 5,525,584; 5,268,378; 5,270,306; 5,342,946; 5,356,902; 5,364,876; 5,376,784; 5,395,827; 5,399,696; 5,407,935; 5,420,155; 5,426,106; 5,446,051; 5,475,008; 5,504,085; 5,510,338; 5,631,373; 5,514,680; 5,519,019; 5,521,174; 5,527,810; 5,532,236; 5,559,106; 5,559,125; 5,580,877; 5,606,062; 5,614,508; 5,614,532; 5,672,705; 5,721,234; 5,639,751; 5,654,303; 5,807,851; 6,060,479; 6,075,018; 6,096,743; and 6,200,999 and International Patent Applications (PCT): WO 92/07847; 93/05772; 94/25469; 95/311443; 95/35289; 96/11922; 96/14318; 96/28445; 96/37500; 97/10246; 97/19066; 97/25326; 97/25327; 97/25329; 97/28163; 97/34878; 99/07708 and European Patent Applications EP 0 315 959 A2; 0 348 872 A1; 0 374 534 A1; 0 377 112 A1;

459,561 A2; 0 503 349 A1; 0 511 152 A2;

676 397 A1; and 0 699 676 A1.

GYKI 52466 [1- (4-Aminophenyl) -4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine] is a non-competitive AMPA antagonist disclosed in U.S. Patent No. 4,614,740. patent. GYKI 52466 has been shown to be a highly selective AMPA receptor antagonist with a non-competitive mode of action, i.e., allosteric action (Donevan and Rogawski, Neuron 10:51 (1993); ibid., 10:61 (1993)]. The non-competitive AMPA antagonist activity of GYKI 52466 as well as its antispasmodic, muscle relaxant and neuroprotective properties have been fully supported by detailed pharmacological studies [Upton, Trends in Pharmacol. Sci., 15: 456 (1994)]. Similarly, the non-competitive antagonist LY 300164 [(-) 1- (4-aminophenyl) -3-acetyl-4-methyl-7,8-methylenedioxy-3,4-dihydro-5H-2,3-benzo diazepine (GYKI-53773, also known as Talampanel)] has been shown to increase the anticonvulsant effect of diazepam and clonazepam and other antiepileptics (Borowitz et al., Eur. J. Pharmacol. 380: 67 (1999), Czuczwar et al., Metab. Brain Dis., 11: 143 (1996)]. The magnitude of the protective effect observed depended on the epileptic model examined. Furthermore, Talampanel has also been shown to improve levodopa-induced dyskinesia in MRTP monkeys (Konitsiotis et al., Neurology 54: 1589 (2000)).

European Patent Application 0548 370 A1 discloses a 2,3-benzodiazepine derivative [1- (3,4-dimethoxyphenyl) -5-ethyl-7,8-dimethoxy-4-methyl-5H-2,3 benzodiazepine], also known as Tofizopam, and inhibits IgE production. Tofizopam has been reported to exert an anxiolytic effect by binding to opioid receptors (Horváth, Pharmazie 56 (1): S56 (2001)). Tofizopam has so far not been shown to modify the AMPA receptor.

To date, effects of AMPA receptor-related effects on topical administration of antagonists and / or modulators, or the physical presence of AMPA receptors in the respiratory tract have not been described in the literature.

Therefore, there is a need to identify and develop beneficial formulations and methods for treating inflammatory diseases of the respiratory tract, particularly in the case of allergic inflammations such as asthma and asthma-related diseases. Identification and development of beneficial formulations and methods for the treatment of inflammatory airways are particularly important in light of the limitations of current causal therapies. In asthma, the most common inflammatory disease of the respiratory tract, currently available treatments are almost exclusively aimed at either eliminating airway inflammation or dilating obstructed airways, while other features of human asthma such as airway hyperreactivity and structural changes (e.g. hypertrophy and / or hyperplasia of smooth muscles and connective tissues; accumulation of collagen and other matrix components in the subepithelial region of the airway wall; Therapeut, 91: 93 (2001)], are largely neglected, as shown by studies of the efficacy of treatments.

Summary of the Invention

The present invention relates to the use of nitrogen-containing heterocyclic compounds known as AMPA receptor antagonists in inflammatory diseases of the respiratory tract3

EN 226 376 Β1, including asthma and asthma-related diseases. Asthma-related diseases may include allergic reactions or inflammatory diseases of the respiratory tract that are no longer treatable by conventional methods, including pathological changes in the respiratory tract.

Thus, the present invention provides methods for treating inflammatory diseases of the airways in a mammal using a therapeutically effective amount of a nitrogen-containing heterocyclic compound of the invention. In embodiments of the present invention, the AMPA receptor antagonist is either a non-competitive or a competitive AMPA receptor antagonist. The pharmaceutical compositions of the present invention are useful for the treatment of allergic inflammatory diseases of the respiratory tract. Some of the typical inflammatory diseases of the respiratory tract include, but are not limited to, non-allergic asthma and allergen-induced bronchial asthma, chronic inflammatory reactions secondary to chronic bronchitis, chronic obstructive pulmonary disease, and pulmonary fibrosis. The treatments of the present invention are also useful for treating other non-allergic inflammatory diseases of the respiratory tract including, but not limited to, idiopathic pulmonary fibrosis and autoimmune lung disease.

Specification

Surprisingly, it has been found that certain nitrogen-containing heterocyclic compounds known as AMPA receptor antagonists are useful in the treatment of inflammatory diseases of the respiratory tract. These AMPA receptor inhibitors reduce bronchial airway hypersensitivity, which is usually associated with airway inflammation. Such compounds of the present invention are also useful for suppressing growth factor-induced proliferation of smooth muscle cells and inhibit allergen-induced mucus secretion by airway epithelial cells, thus providing novel methods for the treatment of airway obstruction. These observations have led to effective and versatile methods of treating inflammatory diseases of the respiratory tract and related pathological conditions.

The present invention provides pharmaceutical compositions and methods for treating the symptoms of inflammatory diseases of the respiratory tract of a mammal. More particularly, the present invention provides compositions and methods for treating allergic airway inflammation, including asthma or asthma-related disorders such as allergic reactions or an inflammatory disease that can no longer be treated with conventional therapies, including, for example, airway disease.

The compositions and methods of the present invention also provide tools for examining the role of compounds having AMPA receptor antagonist properties in the etiology and development of various airway inflammations, including allergic airway inflammation such as asthma and asthma-related diseases, in various experimental models. pathological changes in the respiratory tract.

abbreviations

AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazole propionate. Clonazepam, 7-nitro-5- (2-chlorophenyl) -3H-1,4-benzodiazepin-2 (1H) -one.

Diazepam, 7-chloro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one.

EG1S 8332, 5- (4-Aminophenyl) -7-acetyl-7,8-dihydro-8-cyano-8-methyl-9H-1,3-dioxo [4,5-h] [2,3] benzo benzodiazepine. GYKI 52466, 1- (4-Aminophenyl) -4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine.

GYKI 53405, 1- (4-Aminophenyl) -3-acetyl-4-methyl-3,4-dihydro-7,8-methylenedioxy-5H-2,3-benzodiazepine.

GYKI 53655, 1- (4-Aminophenyl) -3-methylcarbamoyl-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine.

GYKI 53773, (Talampanel. LY 300164), (-) - 1- (4-Aminophenyl) -3-acetyl-4-methyl-3,4-dihydro-7,8-methylenedioxy-5H-2,3- benzodiazepine.

GYKI 53784, (LY 3030701 (-) - 1- (4-Aminophenyl) -3-methylcarbamoyl-4-methyl-7,8-methylenedioxy-3,4-dihydro-5H-2,3-benzo benzodiazepine.

GYKI 47261, 6- (4-Aminophenyl) -8-chloro-2-methyl-11H-imidazo [1,2-c] [2,3] benzodiazepine.

NBQX, 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo (f) -quinoxaline.

SYM 2267, 1- (4-Aminophenyl) -3,5-dihydro-4-methyl-3-acetyl-7-methoxy-5H-2,3-benzodiazepine.

Technical and scientific terms used herein are to be understood in the sense used by one of ordinary skill in the art to whom this invention is written, unless otherwise defined. Various methods and materials known to one of ordinary skill in the art will be incorporated herein by reference. Literary governing labor laws, the laws on general pharmacology for example, Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10 ^th Ed., McGraw Hill Companies Inc., New York (2001). Any suitable material and / or method known to one of ordinary skill in the art may be used in the practice of the present invention. However, preferred materials and methods are provided. Materials, reagents, and the like, which are referred to in the following description and examples, are commercially available, unless otherwise indicated.

The inventors have uncovered the presence of AMPA receptors in the lungs and for the first time demonstrated their unknown role in lung-related diseases. Thus, the present invention provides methods for treating inflammatory airways disease in a mammal using a therapeutically effective amount of a nitrogen-containing heterocyclic compound of the invention known to inhibit the AMPA receptor.

As used herein, the terms "treat" or "treat" mean to reduce, alleviate, prevent, and / or reverse the symptoms of a disease. A disease treatable by the methods and compositions of the invention is any disease

EN 226 376 Β1 weeks characterized by or including acute or chronic respiratory tract inflammation. Thus, the terms "inflammatory disease" or "airway inflammatory disease" include any inflammatory pulmonary disease including asthma, non-allergenic or allergen-induced bronchial asthma, acute or chronic bronchitis, hay fever, chronic bronchitis, and lung disease, pulmonary fibrosis. The present invention is also useful in any pulmonary disease in which white blood cell and airway abnormalities may play a role, including but not limited to idiopathic pulmonary fibrosis and any other autoimmune disease, wherein "asthma" is a disease having a continuous or heavy breathing in seizures, accompanied by wheezing, tightness in the chest and often coughing or stuck breathing attacks. By "asthma-related diseases" is meant a condition whose symptoms are predominantly inflammatory with associated bronchospasm. Thus, both asthma and asthma-related diseases are characterized by symptoms that include airway narrowing due to varying degrees of smooth muscle contraction (spasm), mucosal edema, including mucus of the upper respiratory tract, bronchi, and bronchi. Non-limiting examples of "asthma-related diseases" include non-asthmatic diseases characterized by respiratory hypersensitivity (e.g., chronic bronchitis, emphysema, cystic fibrosis and respiratory pain).

The formulations and methods disclosed herein are examples of use in asthma. However, the invention is not to be construed as limited to this lung disease. Asthma has the advantage of being studied in detail and has many accepted models to support the invention. Sensitization and allergen test are known to lead to airway hypersensitivity in various agonists. Thus, acetylcholine, a known anticonvulsant, is able to induce greater contraction of muscle cells of tissues obtained from the trachea of killed animals (previously sensitized to induce respiratory hypersensitivity) than those isolated from control animals following an allergen challenge. for example, Tokuoka et al., Br. J. Pharmacol., 134: 1580 (2001); Nakata et al., Int. Immunol. 73: 329 (2001); Emala and Hirshman, Monogr. Allergy 33:35 (1996)].

The most prominent feature of asthma is bronchospasm or narrowing of the airways. Asthmatic patients show marked contraction of small and large airways, increased mucus formation and inflammation (Plaut and Zimmerman, supra). In asthma, the inflammatory response is characterized by mucus-coated tissues and by vasodilation, plasma secretion, by the influx of inflammatory cells such as neutrophils, monocytes, macrophages, lymphocytes and eosinophils into the site of inflammation, and by the release of inflammatory mediated respiratory epithelial cells) or inflammatory cell migration (Hogg, "Pathology of Asthma, in Asthma as an Inflammatory Disease, O'Byrne (ed.), Marcel Dekker, Inc., New York, NY, at 1 (1990)). . Asthma can be triggered by a variety of causes such as allergic reactions, secondary responses to infections, industrial or occupational exposure, ingestion of certain chemicals or drugs, and physical exertion [Hargreave et al., J. Allergy Clin. Immunol. 83: 1013 (1986)].

The compounds of the invention are effective in reducing mucus production of bronchial epithelial cells and inhibiting growth factor-mediated smooth muscle cell proliferation.

Bronchial hypersensitivity (AHR), a more severe form of asthma, can be triggered by both respiratory antigen and non-antigenic stimuli. Late response and persistent hypersensitivity in allergic-induced asthma have been linked to the assembly of leukocytes, and in particular eosinophils, in inflamed lung tissue [Abraham et al., Am. Respir. Dis. 138: 1565 (1988)]. Eosinophils release a number of inflammatory mediators, including 15-HETE, leukotriene C4, PAF, cationic proteins, and eosinophil peroxidase.

Endothelins represent another class of anticonvulsants among the most active known vasoconstrictor hormones. A typical endothelin is endothelin-1 (ET-1), a 21 amino acid peptide that is produced in a variety of tissues including endothelial and vascular smooth muscle cells, bronchial epithelial cells, neuronocban and central nervous system astrocytes, and endometrial cells. Bronchial epithelial cells respond to IgE-induced activation by endothelin release, suggesting a direct role of the epithelial layer in the allergic response. Endothelin-1 has many effects including smooth muscle contraction, mitogenesis, microvascular leakage and edema, mucosal hyperproliferation and neuromodulation suggesting its possible role in the pathophysiology of various lung diseases [Id. for example, Sokolovsky, Pharmacol. Ther. 68: 435 (1995); Rubanyi and Polokoff, Pharmacol. Port. 46: 325 (1994); Goldie and Femandes, Monaldi Arch. Chest Dis., 55: 162 (2000)]. ET-1 has been shown to be produced predominantly in the lungs and that lung tissue has a very high receptor density for this peptide (e.g., Hemsen et al., Eur. J. Pharmacol. 191: 319, 1990). Endothelins cause contraction of the lungs' smooth muscles, similar to the effect seen during an asthma attack. Asthma patients are more sensitive to ET-1 bronchoconstriction than non-asthmatics [Chalmers et al., Am. J. Respir. Crit. Care Med. 156: 382 (1997)] and it has also been shown that the ET-1 content of arterial blood and bronchoalveolar lavage fluid is higher in asthma attacks and

HU 226,376 Β1 during the course of the process compared to healthy controls [Trakada et al., Respir. Med. 94: 992 (2000)]. Therefore, inhibition of ET-1 production or activity reduces asthmatic symptoms. Administration of corticosteroids has been associated with a significant decrease in ET-1 levels in bronchial biopsy specimens and bronchoalveolar lavage fluid, and a decrease in ET-1 release from bronchial epithelial cells from asthmatic patients [Mattoli et al., J. Allergy Clin. Immunol. 88: 376 (1991); Redington et al., J. Allergy Clin. Immunol. 100: 544 (1997); Vittori et al., Am. Rev. Resp. Dis. 146: 1320 (1992)]. It has been discovered that the compounds of the invention reduce the contraction of ET-1-induced lung smooth muscle tissue in animals.

The terms "antigen" and "allergen" are used interchangeably to describe molecules, such as dust or pollen, that may trigger an allergic reaction and / or asthmatic symptoms in an individual suffering from asthma, such as an asthmatic subject who is allergic or antigenic. Being treated with a sufficient amount of an allergen or antigen to induce an asthmatic response. The compounds of the invention are effective in the treatment of AHR after ovalbumin sensitization and antigen treatment.

An "AMPA receptor antagonist" means any unit that inhibits AMPA receptor activity. Such an AMPA receptor antagonist may be competitive or non-competitive.

The use of the present invention includes nitrogen-containing heterocyclic compounds and their pharmaceutically acceptable salts, known as AMPA receptor antagonists in Table 1 below:

Table 1

Non-competitive AMPA receptor antagonist compounds

Structure Name Class Literature CH ₃ <XX f HjN GYKI-52466 2,3-benzodiazepine Tamawa. et al., Eur. J. Pharmacol. 167: 193 (1989). / H <XCH HjN racem: GYKI-53405 (-) enant: GYKI-53773 (LY-300164) Talampanel Dihydro-2,3-benzodiazepine Tamawa. et al., Bioorg. Med. Chem. Lett. 3:99 (1993) U.S. Patent No. 5,536,832. patent ch ₃ Ο ^ N- ^CH 3 h ₂ n racem: GYKI-53655 (-) enant: GYKI-53784 Dihydro-2,3-benzodiazepine U.S. Patent No. 5,536,832. patent

HU 226 376 Β1

Table 1 (continued)

Structure Name Class Literature H, C 0? < ex P θ ' ^CH 3 Egis 8332 U.S. Patent No. 5,807,851. patent Dihydro-2,3-benzodiazepine U.S. Patent No. 6,075,018. patent V ) h ₂ n N- Cl · ^ V HjN ! -N GYKI-47 261 Annulled 2,3-benzodiazepine Abraham et al., Bioorg. Med. Chem. 8: 2127-2000 (2000). W099 / 06408 PH ₃ ch ₃ c \ She 0 N - y = N ^CH 3 SYM-2267 3,5-Dihydro-5H-2,3-benzodiazepine W001 / 98280 A2 X H ₂ N

Nitrogen-containing heterocyclic compounds useful in the present invention which are basic in nature may form salts with a variety of inorganic or organic acids. Although these salts must be pharmaceutically acceptable salts for administration to animals, in practice it is often advantageous to first isolate the compound as a pharmaceutically unacceptable salt from the reaction mixture and then simply convert the latter to the free base using a basic reagent. followed by conversion of the free base to the pharmaceutically acceptable acid addition salt. According to Method 60 of the present invention, the base is formed into the acid addition salt by treating the basic compound with a sufficient amount of the selected mineral or organic acid in an aqueous solvent medium or in a suitable organic solvent such as methanol or ethanol. Careful evaporation of the solvent affords the desired solid salt.

The acids that can be used to prepare the pharmaceutically acceptable salt of a nitrogen-containing heterocyclic compound useful in the present invention are non-toxic acid addition salts, such as those containing pharmaceutically acceptable anions (see Remington, pp. 7

Such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate or bisulfate, phosphate or acid phosphate, acetate, lactate, citrate or acid citrate, tartrate or salts of bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate and pamoate (i.e. 1,1'-methylene-bis (2-hydroxy-3-naphthoate)).

It is intended that the methods of the present invention be applicable to any mammal that may experience the benefits of the methods of the present invention. These mammals are primarily human, but the invention is not limited in this sense, that is, it can be used in veterinary medicine. Thus, in accordance with the present invention, the terms "mammal" or "mammal in need of treatment" include both human and non-human mammals, preferably domestic animals including, without limitation, cats, dogs and horses.

The term "therapeutically effective amount" refers to the effective dose regimens at which the desired therapeutic result is achieved. Further, one skilled in the art will recognize that a therapeutically effective amount of any compound of the invention may be reduced or increased by fine-tuning and / or by administering more than one of the compounds of the invention or by co-administering the compound of the invention with another anti-asthma agent. Thus, the present invention provides a method of adjusting dosing / treatment to the individual needs of a particular mammal. As illustrated in the following examples, the therapeutically effective amount can be readily determined, for example, based on experience, using relatively low initial amounts, and gradually increasing, while establishing a beneficial effect. Clinical changes that are important for determining the therapeutic effect of the treatment of the present invention include, but are not limited to, reducing symptoms and signs of asthma and related conditions (e.g., difficulty breathing, wheezing, coughing, bronchial hypersensitivity, pathological changes in the airways) and improving lung functional tests. These are based on the patient's symptoms and the doctor's observation.

As used herein, specifying a range of numbers for a variable expresses that the variable can be any value within this range in the practice of the invention, so that for a variable that is not substantially continuous, the variable can be any integer within this range, including the extreme values of the range. Similarly, for a variable that is essentially continuous, the variable can be any real value within this number range, including the extreme values of the range. For example, a variable that can have a value between 0 and 2, which is essentially non-continuous, can be 0, 1, or 2, or, if it is substantially continuous, can be 0.0, 0.1, 0.01, 0.001, or any other real value.

Therefore, therapeutically effective amounts may range from about 0.01 mg / kg / day to about 300 mg / kg / day when administered systemically (e.g., orally). In a preferred embodiment of the invention, the therapeutically effective amounts when administered systemically are about 0.1 mg / kg / day and

They may range from 150 mg / kg / day.

For topical administration, such as inhalation, therapeutically effective amounts may be in the range of about 0.01 pg / kg / day to 300 pg / kg / day when administered systemically (e.g., by inhalation to the respiratory tract). In a preferred embodiment of the invention, when administered systemically, the therapeutically effective amounts may range from about 0.1 pg / kg / day to about 150 pg / kg / day.

The dosage forms and the frequency with which they are administered will depend on the usual factors considered by one of ordinary skill in the art in determining therapeutically effective amounts for a particular mammal, as discussed above. Thus, the practitioner will consider the condition of the patient being treated, the particular compound of the invention that is being administered, the route of administration, and other clinical factors such as age, weight, and condition of the mammal, as well as comfort and patient agreement.

It will be apparent to one of ordinary skill in the art that the number of administrations of the compounds of the invention will vary from patient to patient at any given time depending on the particular medical condition of the patient.

When applicable (such as in the treatment of asthma), the compound of the present invention may be administered prior to, simultaneously with, or after exposure to the antigen of the mammal. Further, the time of administration of the compound of the present invention will vary from mammal to mammal in relation to the antigenic effect depending on the particular situation. An expert practitioner will optimize dosing by carefully monitoring the patient while changing the time and / or sequence of administration of the compound of the invention. Thus, it is clear that the present invention does not cause the mammal to suffer from certain inflammatory lesions of the lungs. The compounds of the invention may be administered prophylactically to individuals predisposed to asthma and / or asthma-related diseases. For example, a person who is allergic to pollen may administer a compound of the invention (e.g. orally) daily and / or prior to entering a pollen-rich area (e.g., a garden). Similarly, a person with a family history of asthma attack may be given prophylactic compounds of the invention to prevent such an asthma attack from occurring.

The compounds of the invention are preferably formulated with a pharmaceutically acceptable carrier, any known pharmaceutically acceptable carriers, including diluents and excipients (see Remington's Pharmaceutical Sciences, 18 ^th Ed., Gennaro, Mack Publishing Co., Easton, PA 1990 and Remington. The Science and Practice of Pharmacy, Lippincott, Williams & Wilkins, 1995). While the type of pharmaceutically acceptable carrier / carrier used in the preparation of the composition of the invention will depend upon the use of the composition in a mammal.

In general, the pharmaceutically acceptable carriers are physiologically inert and non-toxic. The compositions of the invention may contain several types of compounds of the invention as well as any other active ingredient which is effective in treating the specific pneumonia to be treated. Such compounds include, but are not limited to, β-andrenoceptor antagonists: albuterol, metaproteranol, levealbuterol, pirbuterol, salmeterol, bitolterol; glucocorticoids: beclamethasone, triamcinolone, flunisolide, budesonide, fluticasone; leukotriene receptor antagonists and leukotriene synthesis inhibitors: zafiriukast, montelukast, zileutin; other anti-asthma agents: cromolyn, nedocromil, theophylline; anticholinergics: ipatropium, oxitropium, tiotropium;

H ₁ receptor antagonists antihistamines: diphenhydramine, pyrilamine, promethazine, loratidine, chlorocyclizine, chlorophemiramine, fexofenadine; adrenocorticosteroids; and glucocorticoids.

The compositions of the invention may be administered by conventional routes (e. G., Oral, inhalation, rectal, nasal, topical, including buccal and sublingual routes, or parenterally, including subcutaneous, intramuscular, intravenous, intradermal, transdermal, and intratracheal routes).

In addition, polymers may be blended according to conventional methods known in the art to provide sustained release of a particular compound.

Formulations suitable for inhalation administration may be any which may be practiced on the basis of inhalation inventions known in the art. These formulations may contain carriers such as powders and aerosols. The present invention encompasses liquid and powder formulations suitable for nebulization and intrabronchial use, or aerosol formulations which may be administered by a metered dose aerosol formulation ("MDI"). Particularly preferred embodiments are described in U.S. Patent No. 5,447,150. patent.

The active ingredient may be formulated in an aqueous inhalable pharmaceutically acceptable carrier such as isotonic saline or bacteriostatic water and other carriers of a similar type well known in the art. These solutions may be administered by pump or pressure-driven nebulizer spray 45 or by any other conventional means that will induce or permit inhalation of the required amount of a liquid formulation into the lungs of a patient.

Powder formulations containing the anti-inflammatory compounds of the present invention may be in the form of, for example, lactose or other inert powder which may be a pharmaceutically acceptable powder formulation of a thoroughly mixed active ingredient for intrabronchial administration. The powder formulations may be administered by the aid of an automatic dispenser, including but not limited to an aerosol dispenser, or enclosed in a breakable capsule, which the patient may fit into a device which breaks the capsule and blows the powder with a constant current.

Aerosol formulations for use in the present method typically contain propellants, surfactants, and solubilizers, and can be filled into conventional aerosol dispensers, which are closed with a different metering valve.

Anti-inflammatory compositions for oral administration of the present invention may be formulated as discrete units such as capsules, lozenges, gelcaps, cachets, pills, or tablets, each containing a predetermined amount of the active ingredient in powder or granules; as solutions or suspensions in aqueous or non-aqueous solution; or an oil-in-water liquid emulsion or a water-in-oil emulsion and a large pill, etc. However, administration of the composition of any aspect of the present invention may be accomplished with liquids, suspensions or elixirs, powders, tablets, micronized particles, or osmotic delivery systems.

Formulations of the compositions of the present invention suitable for nasal administration wherein the carrier comprises a solid, coarse-grained powder having a particle size of, for example, 20 to 500 microns, are administered as is customary for snuffing, i.e., by rapid absorption through the nasal cavity. from a container containing a powder that we keep close to the nose. Formulations suitable for nasal administration wherein the carrier is a liquid, such as a nasal spray, aerosol, or nasal drops, comprise an aqueous or oily solution of the compound of the present invention.

Formulations for parenteral administration may be in the form of aqueous or non-aqueous sterile injectable solutions, which may contain antioxidants, stabilizers, buffers, antimicrobial agents, and solubilizing agents, and may be made isotonic with the blood of the patient to be treated; and aqueous or non-aqueous sterile suspensions which may contain suspending and thickening agents.

The following examples are intended to illustrate preferred embodiments of the invention without limiting the scope thereof. It will be recognized or determined by those of ordinary skill in the art without going beyond conventional experimentation, without many equivalents of the materials and procedures disclosed herein.

Example 1

Examination of the presence / role of AMPA receptors in rat lungs The presence / role of AMPA receptors in the lungs of experimental animals was studied in the same model (perfused ventilated rat lung) as used in the study of NMDA receptors by Said et al. Natl. Acad. Sci. (USA) 93: 4688 (1996)]. The lungs of anaesthetized SPRD rats were ventilated and a continuous flow of Krebs solution was passed through it in situ. L-arginine, AMPA and an AMPA antagonist were added directly to the fluid as described in Said et al., Supra. As shown in Table 2, AMPA9

Activation of the Β1 receptor resulted in an acute edema of the lungs, which is respiratory pressure (Paw). ^the pulmonary artery pressure (P _P) and post-treatment the ratio of dry and wet lung weight in prolonging manifested significant. Administration of GYKI-53784 reduced both airway and pulmonary artery pressure.

Table 2

Effect of treatment with AMPA and a non-competitive AMPA receptor antagonist, GYKI 53784, on various parameters of lung function in perfused lungs of SPRD rats

Parameters Management (n) control AMPA (200 μΜ) AMPA (200 μΜ) + 53784 (200 μΜ) ^r AW 99.3 ± 4.9 (8) 183.4 ± 15.6 (7) 110.3 ± 19.1 (4) P <0,001® <0.02 & ^P AP ** 96.0 ± 8.2 (9) 147.9 ± 8.6 (7) 114.0 ± 23.0 (4) P <0.001 ° DIG* W / D ratio * 6.6 ± 0.4 (3) 14.8 ± 3.9 (3) 14.3 ± 5.6 (4) P NS ° DIG*

* ^p aw ^: respiratory pressure after 60 minutes of perfusion as a percentage of pressure at 0 minutes;

P _AP : pulmonary artery pressure after 60 minutes of perfusion as a percentage of pressure at 0 minutes; W / D: ratio of wet to dry lung weight after 60 minutes of perfusion;

a: relative to control;

b; compared to AMPA treatment.

Example 2

Study of the effect of an orally administered non-competitive AMPA receptor antagonist on airway hypersensitivity ex viv

Airway inflammation can lead to bronchial hypersensitivity, which is a common symptom of asthma.

Rats were sensitized to ovalbumin (OA) by subcutaneous injection on day 1 with 0.5 ml OA / Al (OH ₃ ) gel mixture (2 mg OA + 10 g 35 AI (OH ₃ ) / 100 ml saline), followed by on days 14 and 21, another subcutaneous injection [10 mg OA + 10 g Al (OH ₃ ) / 100 ml saline]. On day 28, the animals received GYKI-52466, GYKI-47261 or GYKI-53773 orally (0.3 mg / kg or 3.0 mg / kg) 2 hours prior to 40 antigen treatment. Antigen treatment was performed by inhalation of nebulized ovalbumin (1% antigen solution administered for 1 hour in a TSE inhalation system). The animals were sacrificed 48 hours after antigen treatment, and the tracheas were removed and placed in an organ bath. The excised tracheas were allowed to equilibrate for 30 minutes before measuring the tracheal response curves induced by acetylcholine (Ach).

As shown in Table 3, ovalbumin treatment of sensitized animals in this model caused significant tracheal hypersensitivity when the response to the anticonvulsant was determined 48 hours after antigen treatment. Administration of GYKI-52466, GYKI-47261 and GYKI-53773 at a dose of 3.0 mg / kg restored this increase to control levels.

Table 3

Effect of antigen treatment and oral pretreatment with various GYKI compounds on acetylcholine-induced tracheal contraction in BN rats

Parameters control Handled GYKI-52466 3.0 mg / kg GYKI-47 261 3.0 mg / kg GYKI-53773 3.0 mg / kg ed ₅₀ ' 5.63 ± 0.11 6.74 ± 0.32 5.60 ± 0.46 P 0,002 0,028 MAX 100 ± 0 276 ± 47 135 ± 32 P <0.001 0.037 ed ₅₀ * 5.22 ± 0.14 5.89 ± 0.18 5.26 ± 0.30 4.64 ± 0.34 P 0,003 0,047 <0.001 MAX 100 ± 0 163 ± 18 98 ± 13 85 ± 16 P <0.001 0,020 0,007

'-Log M acetylcholine (Ach), which causes 50% contraction compared to control (mean ± SEM). Contraction at maximum Ach concentration relative to control (mean ± SEM).

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Example 3

Examination of the effect of a topically applied non-competitive AMPA receptor antagonist ex vivo on airway hypersensitivity The procedures of Example 2 were followed to compare the effect of antigen treatment and topical (intratracheal) treatment with GYKI 53773 on Ach-induced tracheal contraction and allergen-sensitive rats. The results presented in Table 4 with GYKI-53773 (Talampanel) clearly show that the test compound strongly reduced allergen-induced airway hypersensitivity at very low (1 pg / kg) dose after a single dose. .

Table 4

Effect of antigen treatment and topical (intratracheal) treatment with a non-competitive AMPA receptor antagonist (Talampanel) on acetylcholine-induced tracheal contraction in BN rats

Parameters control Handled GYKI-53773 0.001 mg / kg GYKI-53773 0.01 mg / kg GYKI-53773 0.1 mg / kg ed ₅₀ ' 5,8910,12 7,2210,24 5.80 ± 0.47 5.73 ± 0.39 5,6810,55 P <0.001 0.011 0,004 0,013 MAX ** 10010 312 199 202 166 113H9 129 130 P 0.011 NST 0,036 0.053

* -logged M acetylcholine (Ach), which causes 50% contraction compared to control (mean ± SEM). ** Contraction at maximum Ach concentration relative to control (mean ± SEM). t Not significant (p> 0.05).

Example 4

Examination of the effect of systemic (intraperitoneal) administration of a competitive AMPA receptor antagonist on airway hypersensitivity ex vivo The procedures of Example 2 were followed to compare the effect of antigen treatment and systemic (intraperitoneal) treatment of NBQX on Achinduced Tracheal Sensitivity and Tracheal Sensitivity rats.

The data in Table 5 show that the competitive AMPA antagonist NBQX strongly reduces both parameters (ED ₅₀ and maximal evoked response) of allergen-induced respiratory hypersensitivity in experimental animals.

Table 5

Antigen treatment and the effect of systemic (intraperitoneal) competitive AMPA receptor antagonist (NBQX) on acetylcholine-induced tracheal contraction in BN rats

Parameters control Handled NBQX 0.15 mg / kg NBQX 1.5 mg / kg NBQX 15 mg / kg ed ₅₀ * 6.16 ± 0.31 7.31 ± 0.37 6.29 ± 0.51 4,9810,80 5,1110,55 P 0,015 NST 0.011 <0.001 MAX ** 100 ± 0 308 134 178 165 115 130 125 132 P <0.001 NST <0.001 <0.001

* -logged M acetylcholine (Ach), which causes 50% contraction compared to control (mean ± SEM). ** Contraction at maximum Ach concentration relative to control (mean ± SEM). t Not significant (p> 0.05).

Example 5

Competitive applied locally (intratracheally) 55

Examination of the Effect of the AMPA Receptor Antagonist on Airway Hypersensitivity Ex vivo The procedures of Example 2 were followed to compare the effect of antigen treatment and systemic (intraperitoneal) NBQX treatment on Ach-60-induced tracheal contractions in allergen-treated and treated rats. The data in Table 6 show that the competitive AMPA antagonist NBQX inhibits allergen-induced airway hypersensitivity in a dose-dependent manner, but it should be noted that it has a significant effect at higher doses only.

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Table 6

Antigen treatment and the effect of a topical competitive AMPA receptor antagonist (NBQX) on acetylcholine-induced tracheal contraction in BN rats

Parameters control Handled NBQX 0.001 mg / kg NBQX 0.01 mg / kg NBQX 0.1 mg / kg ed ₅₀ * 5.68 ± 0.12 6,7010,55 5,8510,59 5,2010,72 3,4410,36 P 0,010 NST NST <0.001 MAX ** 100 ± 0 3271133 3081128 108 128 60 114 P 0,006 NST NST 0,025

* -logged M acetylcholine (Ach), which causes 50% contraction compared to control (mean ± SEM). ** Contraction at maximum Ach concentration relative to control (mean ± SEM). t Not significant (p> 0.05).

Example 6

Investigation of the effect of AMPA antagonists on ET-1-induced tracheal contraction in vitro In order to determine

The effects of 52466 and 53773 (talampanel) on endothelin-1-induced contractions in isolated guinea pig marrow were as follows. The trachea of male Dunkin-Hartley guinea-pigs (350-450 g) was removed, helical strips were made and placed in an organ bath. After equilibration, the strips were contracted with endothelin-1 and, when the contraction reached the plateau, various concentrations of GYKI-52466, -53773 or the appropriate vehicle were added to the chambers. Changes in voltage were expressed as a percentage of the maximal response to 4.5 μΜ histamine at the start of the experiment.

The results obtained, which are shown in Table 7, demonstrate that both GYKI-52466 and the talampanel dose-dependently inhibit ET-1-induced contraction in isolated guinea pig trachea.

Table 7

Effect of AMPA receptor modulators on endothelin-induced tracheal contractions in vitro

Experiment Treatment Contraction% * Mean ± SD (n) significance ** First Excipients 84 ± 8 (5) <0.05 52466 (10 μΜ) 5317 (5) Second Excipients 6618 (5) <0.01 52466 (30 μΜ) 2319 (5) Third Excipients 8418 (5) <0.05 53773 (10 μΜ) 4319 (8) 4th Excipients 6618 (5) <0.01 53773 (30 μΜ) 714 (4)

* Contraction expressed as a percentage of the maximum response elicited by 4.5 μΜ histamine 25 minutes after administration of the AMPA receptor antagonist.

** ANOVA followed by Dunnett's test compared to vehicle treatment.

Example 7

Study of the effect of AMPA receptor antagonists on allergen-induced production of 50 airway epithelial cells

Antigen-sensitized BN rats were orally pretreated with 0.3, 3 and 30 mg / kg GYKI-52466 or GYKI-53773 (Talampanel) 2 hours prior to antigen treatment using a protocol similar to that used for described in Example. Lungs were isolated 48 hours after antigen treatment and samples were fixed in phosphate-buffered 8% formalin. The samples were then routinely treated for histochemistry. Periodic acid-Schiff stained 60 (PÁS) and haematoxylin-eosin repainted, 5 µm thick, epithelial cells of all airways were counted in the whole preparation at 400X magnification. PAS (+) (mucus producing) epithelial cells were expressed as a proportion of total epithelial cells.

As shown in Table 8, allergen treatment stimulates airway epithelial cell mucus production (control vs. treated). Both compounds significantly reduced the number of mucus-producing epithelial cells (PAS (+)), even at the lowest dose used, and showed a dose-dependent effect, particularly on the talampanel.

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Table 8

Effect of oral administration of non-competitive AMPA receptor antagonists on allergen-induced mucosal production of airway epithelial cells in BN rats

Manage% * p-value control 0.55 - Handled 16.04 ± 1.83 <0.001 53773 0.3 mg / kg 12.34 ± 1.39 0.03 3 mg / kg 9.49 ± 1.08 0.03 30 mg / kg 5.02 ± 0.75 <0.001 control 2.36 ± 0.49 - Handled 26.04 ± 2.50 <0.001 52466 0.3 mg / kg 16.09 ± 1.82 0,006 3 mg / kg 15.18 ± 1.68 0,007 30 mg / kg 14.17 ± 1.59 <0.001

* -PAS (+) cells as% of total epithelial cells (mean ± SEM).

Example 8

Effect of AMPA receptor antagonists on growth hormone-induced smooth muscle cell proliferation

Pretreated and continuously cultured rat aortic smooth muscle cells were artificially aged with serum deficiency (0.1% FCS, 48 hours). Proliferation was induced by re-administration of serum (10%) containing the required growth factors and the effect of different concentrations of AMPA receptor antagonists on cell proliferation by measuring the incorporation of ³ H-thymidine.

As shown in Table 9, the tested AMPA receptor modulators significantly inhibited growth factor-induced rat smooth muscle cell proliferation at micromolar concentrations.

Table 9

Effect of AMPA receptor antagonists on growth factor-induced smooth muscle cell proliferation in vitro

Compound Concentration (gg / ml) Inhibition (% of control) RASMC * A7R5 ** 53773 0.03 26 21 0.3 59 43 3.0 70 87 NBQX 0.03 1 NOT*** 0.3 5 NOT 3.0 94 NOT

Rat aorta smooth muscle cells (primarily). ** Aligned smooth muscle cell line (ATCC).

*** Not tested.

similarities

Although the present invention has been described in detail and its typical embodiments are referred to, it will be apparent to one of ordinary skill in the art that various changes and modifications may be made to the present invention without departing from its spirit and scope. Therefore, for example, those skilled in the art will recognize or be able to employ standard experiments to determine that there may be many equivalents of the typical materials and procedures described herein. These identities are intended to be within the scope of the present invention and are protected by the following claims.

Claims (4)

PATIENT INDIVIDUAL POINTS Use of a nitrogen-containing heterocyclic compound selected from the group consisting of: 1- (4-aminophenyl) -4-methyl-7,8-methylenedioxy-5H-2,3-benzoic acid for the treatment of respiratory diseases of mammals. benzodiazepine; 1- (4-aminophenyl) -3-acetyl-4-methyl-3,4-dihydro-7,8-methylenedioxy-5H-2,3-benzodiazepine; 6- (4-Amino-phenyl) -8-chloro-2-methyl-11H-imidazo [1,2-c] [2,3] benzodiazepine; (-) - 1- (4-Aminophenyl) -3-acetyl-4-methyl-7,8-methylenedioxy-3,4-dihydro-5H-2,3-benzodiazepine (talampanel); (-) - 1- (4-aminophenyl) -3-methylcarbamoyl-4-methyl-7,8-methylenedioxy-3,4-dihydro-5H-2,3-benzodiazepine; 1- (4-aminophenyl) -3,5-dihydro-4-methyl-3-acetyl-7-metoxi5H-2,3-benzodiazepine; 5- (4-aminophenyl) -7-acetyl-7,8-dihydro-8-cyano-8-methyl-9H-1,3-dioxo [4,5-h] [2,3] benzodiazepine; 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo (f) -quinoxaline and pharmaceutically acceptable salts thereof. Use according to claim 1 for the manufacture of a medicament for the treatment of allergic airway inflammation. Use according to claim 2, wherein the allergic inflammatory disease of the respiratory tract is one of the following: asthma, allergen-dependent or independent bronchial asthma, acute or chronic bonchitis, chronic bronchitis-related secondary pneumonia reactions, chronic obstructive pulmonary disease and pulmonary fibrosis. The use of claim 1, wherein the respiratory inflammatory disease is idiopathic pulmonary fibrosis or autoimmune lung disease.