MXPA96005153A - Pharmaceutical agents for the treatment of chronic and agu inflammatory diseases - Google Patents

Abstract

This invention relates to a method for the treatment of anti-inflammatory diseases in mammals, particularly in humans, consisting in the administration to said mammals of an anti-inflammatory amount of a compound of the formula (I) or a pharmaceutically acceptable addition salt. acceptable of the same in which R3 and R4 are as defined above

Description

PHARMACEUTICAL AGENTS FOR THE TREATMENT OF CHRONIC AND ACUTE INFLAMMATORY DISEASES This invention relates to the use of certain N-substituted mcotmarnide compounds and their pharmaceutically acceptable acid addition salts for the treatment of chronic and acute inflammatory diseases in mammals, including humans. 01 N-3- -rnetox? Benzyl) -2- C4-fluorofenox? ] mcotin-a ida that +? ene l structural formula and other N-substituted N-nicotine compounds are referred to in U.S. Patent 4,861,891 which was issued on August 29, 1989. Eeta patent refers to the use of such compounds as selective inhibitors of phosphodiesterase +? IV (PÜE IV). This patent is incorporated herein by reference in its entirety. The N-substituted nicotmapuda compounds referred to below as "compounds of formula T", which include N-3-methoxy-benzyl-2-phenoxy-cyanide, are also useful for enhancing the CAMP-induced elevation of prostaglandin. aEi in U-937 cells, inhibition of leukotriene synthesis and release mediator in human eosinophils. These compounds also show a reduced emetic response in ferrets. This invention relates to a method of inhibiting type IV phosphodiesterase or to the production of tumor necrosis factor in mammals, particularly in humans, which comprises administering to said mammal an anti-inflating amount of a compound of formula or a pharmaceutically acceptable acid addition salt camen + e thereof in which R3 is 1-pipepdyl, 1- (3-? ndol? 1) et lo, (C1-C4) alkyl, phenyl, benzyl, 1- (1 - femle + ilo) or benc the unsubstituted ring in which the substrate is chlorine, fluorine, methyl or rnetoxy and said substituent is in the aromatic ring; R * is bicyclo C2.2.1] hept ~ 2-? Lo or a group of formula II wherein Y is hydrogen, fluorine or chlorine; and X is hydrogen, fluorine, chlorine, rnetoxy, trifluoromethyl, cyano, carboxy, methylcarbarnoyl, dimethylcarbamoyl or carboalkoxy (C1-C4). A preferred embodiment of this invention relates to the aforementioned process for the treatment of inflammatory diseases in which the compound administered is one in which R3 is 1-Δ-urea or 1- (3-indolyl) ethyl. Another preferred embodiment of this invention relates to the previously preferred embodiment wherein, in the compound being administered, R3 is l- (3-? Ndol? L) ethyl and R * is b cycle [2.2.1.] hept-2-? Another preferred embodiment of this invention relates to the preferred embodiment above wherein, in the compound to be administered, R * wherein Y is hydrogen and X is carboalkoxy (C1-C4). Another preferred embodiment of this invention < It refers to the preferred embodiment above in which, in the compound being administered, R3 is l-p? per? d? lo and R4 is Another preferred embodiment of this invention relates to the above method for the treatment or prevention of a disease selected from the group consisting of asthma, chronic bronchitis, atopic dermatitis, urticaria, allergic rhinitis, allergic conjunctivitis, vernal conjunctivitis, cosionophilic granuloma, psoriasis, rheumatoid arthritis, septic shock, ulcerative colitis, Crohn's disease with myocardial and brain reperfusion damage, chronic glomerulonephritis, endotoxic shock, respiratory distress syndrome in adults, diabetes insipidus, multiple sclerosis and central nervous system disorders, as well as other diseases that involve the production of TNF, consisting in the administration to the patient of an effective amount of a compound of formula or a pharmaceutically acceptable acid addition salt thereof in which R3 is 1-piperidyl, 1- (3-? ndol? Det? lo, alkyl (Ci-C?., phenyl, benzyl, L- (1 -femlethyl) or unsubstituted benzyl in which the substituent is chloro, fluoro, methyl or rnetox and said substituent is in the aromatic ring; R *, is bc clo [2.2.1] hept-yl or? n group of formula wherein Y is hydrogen, fluorine or chlorine; and is hydrogen, fluorine, chlorine, ethoxy, trifluoromethyl, cyano, carboxy, methylcarbamolium, dimethylcarbamoyl or carboalkoxy (Ci-C *). The compounds of formula I and their pharmaceutically acceptable acid addition salts can be prepared as described in the aforementioned U.S. Patent 4,851,891. Inhibitors of type IV phosphodiesterase are useful for the treatment of a variety of inflammatory and allergic diseases including: asthma, chronic bronchitis, atopic dermatitis, urticaria, allergic rhinitis, allergic conjunctivitis, vernal conjunctivitis, cosynophilic granuloma, psoriasis, reurnatoid arthritis, septic shock, ulcerative colitis, Crohn's disease, damage by reperfusion of myocardium and brain, chronic erulonephritis, endotoxic shock, respiratory distress syndrome in adults. In addition, PDE IV inhibitors are useful for the treatment of diabetes insipidus, multiple sclerosis, and central nervous system disorders such as depression and multi-infarct dementia. It has been shown that inhibitors of TV type phosphodiesterase inhibit the production of tumor necrosis factor. The compounds of formula I are also useful for the treatment of viral infections, in which the viruses are sensitive to upregulation by TNF or cause the production of TNF m alive. The viruses contemplated here for treatment are those that produce TNF as a result of an infection, or those that are sensitive to inhibition, as such by decreasing the replication, directly or indirectly, by TNF inhibitors of the T-formula. These viruses include, but are not limited to, HIV-1, HIV-2 and HIV-3, cytomegalovirus (CMV), influenza, adenovirus and the Herpes group of viruses, such as, although not lipidated, Herpes zoster and herpes sirnplex. . This invention relates more specifically to a method for the treatment of mammals, affected by the human immunodeficiency virus (HIV), comprising the administration to said mammals of an effective TNF inhibiting amount of a compound of formula i. The compounds of formula I can also be used in association with the veterinary treatment of animals, in addition to humans, when inhibition of TNF production is necessary. The treatment of TNF-mediated diseases, therapeutically or prophylactically, in animals includes disease states such as those mentioned above, but in particular viral infections. Examples of such viruses include, but are not limited to, the virus of feline immunodeficiency. (IVF) or other retroviral infections such as equine infectious anemia virus, caprine arthritis virus, visna virus, aedi virus and other lentiviruses. The viruses of formula I are also useful for the treatment of infections caused by yeasts and fungi, in which such yeasts and fungi are sensitive to upregulation by TNF or will cause the production of live TNFm. A preferred disease state for treatment is fungal meningitis. In addition, the compounds of formula T can be administered together with other drugs of choice for systemic infections caußadaß by yeasts and fungi. The drugs of choice for physical infections include, but are not limited to, compounds called polymyxins, such as Polymycin B, to the class of compounds called irnid zoles, such as clot rirnazole, econazole, miconazole, and ketoconazole; to the class of compounds called tpazoles, such as co-fluconazole, and itranazole, and to the class of compounds called Arnfotericmas, in particular-Ainfotericin B and ßmfotericin B lLósórnica. The co-administration of the anti-fungal agent with a compound of formula I can be in any preferred composition in which the compound as such is well known to all those skilled in the art., for example various formulations of Arnfotericina B. The coadrninisiation of an antifungal agent with a compound of formula T can be carried out by simultaneous administration or in practice, by separate administration of the agents to the mammals but consecutively. In particular, the compounds of the formula I can be coad administered with an Amphotericin B formulation, especially for systemic fungal infections. The preferred organism for treatment is the Candida organism. The compounds of formula I can be coadministered in a similar manner with antibacterial or antiviral agents. The compounds of formula I can also be used for the inhibition and / or reduction of the toxicity of an anti-fungal, antibactepane or antiviral agent, by administering an effective amount of a compound of formula I to a mammal in need of such treatment. Preferably, a compound of formula I is administered for the inhibition or reduction of the toxicity of the class of compounds of the Amphotericin, in particular of the Amphotericin B. The methods of this invention comprise the administration of compounds of the formula I and their pharmaceutically acceptable acid addition salts and solvates of said compounds and their salts (hereinafter collectively referred to as "or" therapeutics can be administered to said mammals either alone or, preferably, in combination with pharmaceutically acceptable carriers. or diluents in a pharmaceutical composition, in accordance with standard pharmaceutical practice This administration can be carried out by means of various routes, including oral, parenteral and topical, parenteral administration, as used herein, includes but is not is limited to intravenous, intramuscular, intraperitoneal, subcutaneous, and transdermal administration. Generally, administration of the therapeutic agents orally is preferred. For use in the treatment of chronic and acute anti-inflammatory diseases, the most desirable administration of the therapeutic agents, according to this invention, ranges from about 5 mg to about 250 mg per day, preferably about 20 mg up to approximately 120 g per day, in a single dose or in divided doses. For intranasal or inhalant administration, the dose is usually formulated as a 0.1 to 1% solution (p >; / v). In practice, the doctor will determine the actual dose that will be the most appropriate for the patient in particular, which will vary depending on the patient's age, weight and specific response. The above doses are examples of average cases but, of course, there may be individualized cases that merit higher or lower dose ranges, and that these doses are within the scope of this invention. In some cases, it may happen that dosage levels below the lower limit of the above dosage ranges are more than adequate, while in other cases even higher dosages may be used without causing any damage or may be used. produce harmful side effects, provided that these higher dosage levels are first divided into smaller doses that will be administered throughout the day. For purposes of oral administration, tablets containing excipients such as sodium citrate, calcium carbonate and dicalcium phosphate can be used with various disintegrants such as starch and preferably tapioca or potato starch, alginic acid and certain complex silicates, together with binding agents. such co or pol vi ilpyrrolidone, sucrose, gelatin and gum arabic. Furthermore, lubricating agents such as, but not limited to, magnesium etherate, sodium laupl sulfate and talc are often very useful in the production of tablets. Solid compositions of a similar type can also be used as filling of hard and soft elastic gelatin capsules. Preferred materials in this respect may include, by way of example and not limitation, lacrose or milk sugar and also high molecular weight polyethylene glycols. If an oral administration by elixirs and / or suspensions is desired, the essential active ingredient may be combined with various sweetening or flavoring agents, coloring or coloring matters and, if desired, with ernulsifying and / or dispersing agents, together with such diluents such as water, ethanol, propylene glycol, glycepine and various combinations thereof.

For the purposes of parenteral administration, solutions of a therapeutic agent in sesame or peanut oil or in aqueous propylene glycol, as well as in sterile aqueous solutions of the corresponding water-soluble basic salts enumerated above may be employed. These aqueous solutions should be adequately buffered if necessary, and the liquid diluent provided isotonic, with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular and subcutaneous injection. In this relationship, the sterile aqueous medium employed is easily obtained by standard techniques well known to all those skilled in the art. For example, distilled water is commonly used as a diluent and the final preparation is passed through a suitable bacterial filter such as a synthesized glass filter or diatomaceous earths or an unvarnished porcelain filter. Preferred filters of this type include the Berkefeld, the Charnberland and the Asbestos Disfc-Metal Seitz, in which the fluid is aspirated onto a sterile container with the aid of a suction pump. All necessary steps must be taken throughout the preparation of these injectable solutions in order to ensure that the final products obtained are sterile. For transdermal administration, the dosage form of a particular therapeutic agent may include, by way of example, solutions, lotions, ointments, creams, gels, suppositories, formulations and sustained release devices. These dosage forms comprise the particular compound and can include ethanol, water, penetration enhancers and inert carriers such as gels-producing materials, mineral oil, emulsifying agents, benzyl alcohol and the like. In the publications of European Patent Applications 271,983 and 331,382 published on June 22, 1988 and September 6, 1989, respectively, compositions that improve the specific transdermal flow are disclosed. These applications are incorporated herein by reference in their entirety. The ability of the compounds of formula I to inhibit PDE IV and exhibit reduced emetic activity can be determined, respectively, as described in the following protocols.

Inhibition of human type IV phosphodieserase (PDEIV) RECEIVe reconfmani es have been expressed in a Baculov? Rus /? F-9 cell system (Pollok B, Fuog E, Robbins M, Fisher D, Uinland 3, Pillar J, and Cheng 3: "Baculovirus expression of human phosphodiesterase. (PDE) TV isoenzymes: cornparison Uith rPDE-IV produced in mammalian lan and bacterial expression system. Baculovirus and Insect Cell Gene Expression Conference", Pmehurst, NC, March 26-30, nineteen ninety five). The used cell SF-9 was able to hydrolyze cyclic AMP (cAMP) and had no effect on cyclic GMP (cGMP). Kinetic analyzes of these enzymes indicated that the Michaelis constant, Km, to hydrolyze the cAMP was in the range of 0.5 to 4 uM. The enzyme was senessable to nagneeium chloride, and was inhibited by the standard PDEIV inhibitor, rolipra. These preparations were used to evaluate the ability of the test compounds to inhibit PDEIV. Preparation of the test compounds: The compounds were dissolved in methyl sulfoxide at a concentration of 10-2 M, then diluted in water 1:25 (compound 4 x 10-M, 4% rnethyl sulfoxide). Subsequently, serial dilutions were made in methyl. Sulfoxide 4% until reaching the desired concentrations. The final concentration of methyl sulfoxide in the test tubes was 1%. It was added, in triplicate, to 12 x glass tubes 74 rnm, and in this order, and at 4 ° C the following (all concentrations are expressed as final concentrations in the test tube): 25 μl of compound or methyl sulfoxide (1%, for control and blank). 25 ul assay buffer (50 rnM Tris, 10 mM magnesium chloride, pH 7.5) 25 μl [3 H] -cAMP (1 μM) 25 μl of PDEIV enzyme (for the blank, the enzyme is preincubated in a boiling water bath for 10 minutes). The reaction tubes were shaken and placed in a water bath (37 ° C) for 10 to 30 minutes, at which time the reaction was stopped by placing the tubes in a boiling water bath for 2 minutes. Wash buffer (0.5 mL, 0.1 M HEPES / 0.1 M sodium chloride, pH 8.5) was added to each tube in an ice bath. The contents of each tube were applied to an Affi-Gel 601 column (boronate affinity gel, 1.2 rnL bed volume) previously equilibrated with the wash buffer. The [3H] cAMP was washed with 2 x 6 mL of wash buffer and the [3H] 5'cAMP was eluted with 6 mL of 0.25M acetic acid. After stirring, 1 mL of the elution was added to the elution. 3 L of atomic light scintillation fluid in a suitable vial, agitated, and equivalent to [3H].

Average of the compound (test compound) Average of the compound (white (enzyme boiled)% Inh? B? Cion = 1- [] X 100% Compound average (control (without compound) - Compound average (white (boiled enzyme) IC50 is defined as the concentration of compound that inhibits 502 hydrolysis of [3H] cAMP to [3H] 5'AMP. In some experiment, the test compounds were evaluated for PDEIV inhibition by the use of a one-step enzyme assay that was similar to the method disclosed by Thompson et. to the. (Thompson 3U, Bro ker G., and Applernan MM: "Assay of cyclic nucleotide phosphodiesterases with radioactive substrates." Methods m Enzymology 38: 205-212, 1974).

Evaluation of cyclic AMP compound elevation (cAMP) Continuous culture growth of human U937 cells was obtained and they were spun at 1400 epm in a Sorvall RT6000B at 2 ° C for 5 minutes. The supernatant was decanted, and the cell pellet was resuspended in the RPMI 1640 cell culture medium plus 2% fetal bovine kidney (hereinafter FBS). The cells were counted and their viability was checked using a hernocytometer. An appropriate volume of RPMI + 2% FBS was added to the cell suspension to obtain a cellular concentration of 10 * cells per rnL. 500 μL (5 x 10 5 cells) of the suspension was added to 12 x 75 mm glass tubes containing +/- the test compound, in duplicate or in triplicate. This mixture was allowed to incubate at 37 ° C for 1.5 minutes. Added n? L of PGE1 and allowed the incubation to proceed for another 15 minutes, at which time the tubes were placed in a boiling water bath for 10 minutes. The tubes were rotated for 10 minutes at 3700 rpm in the Sorvall RT6000B. A sample of 10 μL of supernatant was carefully made to be subjected to a radioimmunoassay (hereinafter RIA). An RIA cAMP test kit (new England Nuclear cat. No. NEK-033) was used to quantify the amount of cAMP of each tube. The instructions were followed according to the procedure for a non-acetylated enema as described in the RIA manual provided with the kit. The values of cAMP were corrected according to the dilutions and the cell concentration was expressed in piconols (pmol) of cAMP produced by 1 x 10 * cells. The average value of each sample +/- the standard error (if applicable) was also calculated. Any of the two measures of compound potency (EC50 or PC50) were calculated ~ EC50 is the concentration of the compound q? E produces 50% of the arbitrary maximum response (ie, what concentration of cAMP is produced by 10 μM of roliprarn) after subtracting the response from the baseline. PC50 is the concentration of the compound that produces 50% of the maximum response of this compound after subtracting the response from the baseline. The maximum response of a test compound must be at least 150% of the response of the baseline in order to be calculated, the PC50.

Inhibition of EDN / LTE compound production * 100 mL of healthy donor blood was obtained in Vacutamer tubes # 6480 (143 units USE of sodium heparin, 10 mL). The heparinized whole blood was pooled in a silicolated glass beaker or in 50 mL centrifuge tubes at room temperature. A complete blood smear was performed using the Diff-Guik method to determine the percentage of eosirophils relative to total rnonon-clears leukocytes. 1 rnL of whole blood was placed in siliconized or propylene 12 x 75 μm glass tubes containing 1 μL of methyl sulfoxide or 1 μL of the 1000X test compound in triplicate. After stirring, the tubes were placed in a shaking water bath at 37 ° C for 15 minutes. L μL of P6E1 in meth l-sulfoxide was added to all tubes to obtain a final concentration of 1 μM or 0.1 μM of PGE1. After the turmoil, LOO μL of PBS (negative control) or Sephadex G-15 beds in PBS (final concentration of 8.25 to 15.5 rng / rnL) were added to the tubes. After shaking all the tubes were incubated in a shaking water bath at 37 ° C for 1 to 2 hours. At the end of the incubation, 20 μL of 15% ethylene diamine raacetic acid in PBS was added to each test tube to obtain a final concentration of 0.3%. After stirring, all the test tubes are centrifuged at 2000 rpm at 22 ° C for 5 minutes. All plasma supernatants were analyzed for EDN levels. An RIA EPX test was carried out following the instructions of the case manufacturer (Kabi Pharmacia Diagnostic) with all volumes 5 times lower than recommended (ie, 10 μL volume of the unknown sample). It has been shown that EDN and EPX are proteins (Dahl et al., "Asthma: Basic Mechanisms and Clinical Management 2nd ed., Page 112, 1992.) EDN levels are calculated by comparison with a standard curve using Microsoft Excel. or other suitable computer programs and are expressed as ng / mL. The percentage of EDN control release is calculated using the following equation:% EDN control- (EDN sample - white EDN) / (total EDN - white EDN) in which EDN White is the release of EDN in the absence of Sephadex 6-15 and total EDN is the release of EDN in the presence of Sephadex G-15. When appropriate, the IC30 will be determined for the compounds of use using Microsoft Excel - other suitable computer programs.

Immunoassay of LTE4 enzyme (ETA) All plasma supernatants were analyzed for LTE4 levels. An EIA LTE4 assay was carried out following the instructions of the case manufacturer (Cayman Chemical) using a dilution of the 1:10 plasma sample in EIA buffer as unknown. Other LTE4 patterns different from those provided with the case (for example, Biomol) may be used. The LTE4 levels were calculated by comparison with a standard curve using Microsoft Excel or other suitable software and expressed as ng / mL. The percentage of LTE4 release is calculated using the following equation:% LTE4 control =) LTE4 sample - white LTE4) / (total LTE4 - white LTE4) where white LTE4 is the release of LTE4 in the absence of Sephadex G-15 and Total LTE4 is the release of I.TE4 in the presence of Sephadex G-15. Where appropriate, the IC50 for the test compounds will be determined using Microsoft Excel or other suitable software.

Evaluation of the emetic activity of the compounds The test compound was dosed to 5 male ferrets (either oral or i.p.) and placed in individual plexiglass cages. The vehicle for oral dosing was 2% Tween 80 in 3 mL of distilled water. The vehicle for intraperitoneal administration was 3 mL of distilled water. The animals were observed continuously throughout the study period (usually 60 minutes). Behaviors were recorded according to the following markers: (1) vomiting production, (2) arcades (non-productive rhythmic concentrations versus closed glottis), (3) nausea, in which the animal becomes fatigued in front of a closed glottis with mouth open (this does not include contraction of rhythmic abs). The number of each behavior and the time in which the emesis took place was recorded for each animal. Other behaviors can also be noted. The huronee were not re-studied for 7 days. The study with the PDEIV inhibitor, rolipra, indicated that the bacteria could be exposed weekly to this emetic stimulus 3 times without showing emesis to the saline in week 4. After 4 weekly expoeicionee to the rolipram, the ferrets showed an anticipated emesis saline in week 5. Therefore, the animals used in this trial were only exposed to the emetic stimulus 3 times and then removed from the colony. If the plasma drug levels were required, the animals were anesthetized with Ro purn and Ceta ina and 1 L of blood was extracted from the heart using conventional cardiac puncture techniques. The data were expressed as number of vomits, retching and nausea by groups of 5 animals. The ratio of animals that showed vomiting, retching or nausea to the total number of animals was calculated as [Number of emesis samples / Total number in the eneay group) x 1.00].

Claims (6)

NOVELTY OF THE INVENTION CLAIMS

1. - The use of a compound of formula or a pharmaceutically acceptable acid addition salt of the mole in which R3 is 1 -? pepd? lo, l- (3-mdol? l) et? lo, (C 1 -C 4) alkyl, phenyl, benzyl, 1- (1-phenylethyl) or monosubstituted benzyl in which the substituent is chloro, fluoro-methyl or r-methoxy and said constituent is in the aromatic ring; R * is bicyclo [2.2.1] he? T-2-? Or o? N group of formula wherein Y is hydrogen, fluorine or chlorine; and X is hydrogen, fluorine, chlorine, methoxy, trifluoromethio, cyano, carhoxy, netilcarbamoyl, dimethicarcarnoyl or carboalkoxyl (Ci-C4), in the preparation of compositions for the inhibition of type IV phosphodiesterase or the production of the factor of tumor necrosis in mammals, particularly in humans.

2. The use of a compound according to claim V 1, further characterized in that the compound that is used is one in which R 3 is 1-piperidyl, 1- (3-indolyl) ethyl.

3. The use of a compound according to claim 1, further characterized in that the compound used is one in which R3 is l- (3-indolyl) ethyl and R * is bicyclo [2.2.1. ] hept-2-yl.

4. The use of a compound according to claim 1, further characterized in that the compound that is used is one in which R * ee wherein Y is hydrogen and X is carboalkoxy (C? -C ").

5. The use of a compound according to claim 1, further characterized in that the compound q? E is used is one in which R3 is 1-? I? Eridyl and R? Is

6. The use of a compound according to claim 1, further characterized in that the composition obtained is used for the treatment or prevention of a disease selected from the group consisting of asthma, chronic bronchitis, atopic dermatitis, urticaria, allergic rhinitis, Allergic conjunctivitis, vernal conjunctivitis, cosinophilic granuloma, psoriasis, rheumatoid arthritis, septic shock, ulcerative colitis, Crohn's disease with myocardial and brain reperfusion damage, chronic glomerelonephritis, endotoxic shock, respiratory distress syndrome in adults, diabetes insipidus , multiple sclerosis and alterations of the central nervous system, as well as other diseases that involve the production of TNF, consisting of the adninietration to the patient of an effective amount of a compound of formula or a pharmaceutically acceptable acid addition addition of the residue in which R3 is 1-piperidyl, 1- (3-indolyl) etiio, (C1-C4) alkyl, phenyl, benzyl, 1- (1-phenylethyl) or non-substituted benzyl wherein the substituent is chloro, fluoro, methyl or methoxy and said solvent is in the aromatic ring; R * is bicyclo [2.2.1] hept-2-yl or a group of the formula II wherein Y is hydrogen, fluorine or chlorine; and X is hydrogen, chlorine fluorine, methoxy, trifluoromethyl, cyano, carboxy, methyl carbamoyl, di-ethylcarbamoyl or carbo-alkoxy (Ci-C ").