SYNERGIC PHARMACEUTICAL COMBINATION OF A LEUKOTRIENE-RECEPTOR ANTAGONIST AND AN INVERSE AGONIST OF HISTAMINE HI

Abstract

The present invention refers to a pharmaceutical combination comprising the synergic combination of a leukotriene-receptor antagonist such as the active principle montelukast and pharmaceutically acceptable salts thereof, and an histamine H1 inverse agonist such as the active principle ketotifen and pharmaceutically acceptable salts thereof, formulated with pharmaceutically acceptable excipients and/or carriers and/or additives, in a single dosing unit to be administered orally, transdermally or in devices for oral or nasal inhalation, indicated for the treatment of an atopic disease such as asthma, allergic rhinitis and atopic dermatitis.

Claims

1. A synergic pharmaceutical combination comprising: i. a leukotriene-receptor antagonist agent and/or pharmaceutically acceptable salts thereof, ii. a histamine H1 inverse agonist agent and/or pharmaceutically acceptable salts thereof, iii. a pharmaceutically acceptable carrier and/or excipient and/or additive, formulated as a single dosing unit for oral, nasal, intravenous, or intramuscular administration, for treatment and control of an atopic disease in a mammal comprising asthma, allergic rhinitis and atopic dermatitis.

2. The combination of claim 1 wherein the leukotriene-receptor antagonist agent is montelukast or pharmaceutically acceptable salts thereof in a concentration of approximately 0.01 mg/kg to approximately 1000 mg/kg.

3. The combination of claim 1 wherein the histamine H1 inverse agonist agent is ketotifen or pharmaceutically acceptable salts thereof in a concentration of approximately 0.01 mg/kg to approximately 1000 mg/kg.

4. The combination of claim 1 wherein the leukotriene-receptor antagonist agent is montelukast or pharmaceutically acceptable salts thereof in a concentration of approximately 0.01 mg/kg to approximately 1000 mg/kg, and the histamine H1 inverse agonist agent is agent ketotifen or pharmaceutically acceptable salts thereof in a concentration of approximately 0.01 mg/kg to approximately 1000 mg/kg.

5. The combination of claim 1 wherein the single dosing unit is administered orally as a capsule, tablet, lozenge, sublingual tablet, granules, caplet, suspension or solution; or wherein the single dosing unit is an intramuscularly or intravenously injectable formulation; or wherein the single dosing unit is formulated for use in oral or nasal inhalation devices.

6. The combination of claim 1 wherein the mammal is a human.

7. A method of treating an atopic disease comprising administering to mammals suffering the disease an effective amount of a combination of at least one leukotriene-receptor antagonist agents and at least one histamine H1 inverse agonist agents.

8. The method of claim 7 wherein the combination is administered orally, nasally, intramuscularly, intravenously, or a combination thereof.

9. The method according to claim 8 wherein the leukotriene-receptor antagonist agents is montelukast or pharmaceutically acceptable salts thereof administered in an amount of approximately 0.01 mg to approximately 1000 mg/day, and the histamine H1 inverse agonist agents is ketotifen or pharmaceutically acceptable salts thereof administered in an amount of approximately 0.01 mg to approximately 1000 mg/day.

10. The method of claim 7 wherein the leukotriene-receptor antagonist agent is montelukast or pharmaceutically acceptable salts thereof, and the histamine H1 inverse agonist agent is agent ketotifen or pharmaceutically acceptable salts thereof.

11. The method of claim 10 wherein the montelukast or pharmaceutically acceptable salts thereof is in a concentration of approximately 0.01 mg/kg to approximately 1000 mg/kg, and ketotifen or pharmaceutically acceptable salts thereof is in a concentration of approximately 0.01 mg/kg to approximately 1000 mg/kg.

12. The method of claim 7 wherein the atopic disease is asthma, allergic rhinitis, or atopic dermatitis.

13. A method of treating asthma comprising administering to mammals suffering from asthma an effective amount of a combination of at least one leukotriene-receptor antagonist agents and at least one histamine H1 inverse agonist agents.

14. The method of claim 13 wherein the combination is administered orally, nasally, intramuscularly, intravenously, or a combination thereof.

15. The method of claim 14 wherein the leukotriene-receptor antagonist agent is montelukast or pharmaceutically acceptable salts thereof, and the histamine H1 inverse agonist agent is agent ketotifen or pharmaceutically acceptable salts thereof.

16. The method of claim 15 wherein the montelukast or pharmaceutically acceptable salts is administered in a concentration of approximately 0.01 mg/kg to approximately 1000 mg/kg, and the histamine H1 inverse agonist agent is agent ketotifen or pharmaceutically acceptable salts administered in a concentration of approximately 0.01 mg/kg to approximately 1000 mg/kg.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0034] FIG. 1. Dose-response curve of LTD4-induced bronchoconstriction.

[0035] FIG. 2. Dose-response curve of histamine-induced bronchoconstriction.

[0036] FIG. 3. Antagonistic effect of LTD4-induced bronchoconstriction response in the presence of different cumulative concentrations of montelukast (0.003 to 30 IM). The control response is a vehicle.

[0037] FIG. 3. Antagonistic effect of histamine-induced bronchoconstriction response in the presence of different cumulative concentrations of ketotifen (0.1 to 100 μM). The control response is a vehicle.

[0038] FIG. 5. Dose-response relation of Montelukast combined with ketotifen and the relaxation of the bronchoconstriction effect induced by a histamine and LTD4 challenge.

[0039] FIG. 6. Isobologram analysis of the bronchodilation effect of the montelukast-ketotifen interaction with different pharmacokinetic actions. The result is an interaction index of 0.35.

DETAILED DESCRIPTION

[0040] Presently the treatment of asthma is complex, since it involves a specific process including: regular clinical follow-up, education on self-control, measures to avoid triggering factors and of course pharmacologic treatment, which is insufficient, its efficacy is unpredictable, dosing can be cumbersome and adverse effects are common. In the present invention, preclinical tests have proved that particular doses of the novel montelukast and ketotifen combination exhibit an unexpected and vigorous synergic therapeutic effect in the treatment of atopic diseases such as asthma, allergic rhinitis and atopic dermatitis. For this reason, the main object of the present invention is developing a pharmaceutic composition comprising the combination of a leukotriene receptor antagonist. i.e. the active ingredient montelukast and pharmaceutically acceptable salts thereof, and a histamine H1 inverse agonist, i.e. the active ingredient ketotifen and a pharmaceutically acceptable salt thereof, administered with pharmaceutically acceptable additives and/or excipients and/or adjuvants, formulated in a single dosing unit to be orally administered as a solid or liquid, indicated for the treatment and control of atopic diseases such as asthma, allergic rhinitis and atopic dermatitis.

[0041] Presently, an alternative to increase the efficacy of an analgesic treatment and to significantly reduce the secondary effects is through the combined administration of two or more active agents, such as the synergic pharmaceutical combination intended to be protected by the present invention. The montelukast and ketotifen combination has been designed for controlling symptoms, preventing the risk of future exacerbations and achieving the optimal function of the lung as the main organ, with minimal adverse or secondary effects caused by the pharmacological treatment.

[0042] The present invention intends to offer a new therapeutic option for the treatment and control of atopic diseases such as asthma, allergic rhinitis and atopic dermatitis, capable of reducing the symptomatology and improving the patient quality of life. The combination of the aforementioned active principles results in a greater pharmacological power.

[0043] The treatment for asthma and acute respiratory attack must be individualized, stepped-up or -down and continually adjusted in order to allow the patient in need of treatment to obtain a better control of the disease while minimizing adverse effects. This invention is characterized by the fact that the administration of the combination can be stepped-up for a period of time along with clinical visits and self-control (an important responsibility of the patient) in order to avoid a poor evolution (an incorrect evolution can be a consequence of inadequate compliance with the treatment, triggering factors or the presence of other diseases).

[0044] Representatively, at least one leukotriene-receptor antagonist or at least one histamine H1 inverse agonist are given to the patient for this treatment. The leukotriene-receptor antagonist is selected from the group consisting of albuterol sulphate, aminophylline, amoxicillin, ampicillin, astemizole, attenuated tubercle bacilli, azithromycin, bacampicillin, beclomethasone dipropionate, budesonide, bupropion hydrochloride, cefaclor, cefadroxil, cefixime, cefprozil, cefuroxime axetil, cephalexin, ciprofloxacin hydrochloride, clarithromycin, clindamycin, cloxacillin, doxycycline, erthromycin, ethambutol, fenotherol hydrobromide, fluconazole, flunisolide, fluticasone propionate, formoterol fumarate, gatifloxacin, flu virus vaccine, ipratropium bromide, isoniazid, isoproterenol hydrochloride, itraconazole, ketoconazole, ketotifen, levofloxacin, minocycline, montelukast, moxifloxacin, nedocromil sodium, nicotine, nystatin, ofloxacin, orciprenaline, oseltamivir, oseltamivir sulphate, oxtrifilin, penicillin, pirbuterol acetate, pivampicillin, pneumococcal conjugate vaccine, pneumococcal polysaccharide vaccine, prednisone, pirazinamide, rifampin, salbutamol, salmeterol xinafoate, cromoglycate sodium, terbutaline sulphate, terfenadine, theophylline, triamcinolone acetonide, zafiriukast, and zanamivir, while the histamine H1 inverse agonist can be selected from the group consistintg of azelastine, acrivastine, cyclizine, carebastine, cyproheptadine, carbinoxamine, doxylamine, dimetindene, ebastine, epinastine, efletirizine, ketotifen, levocabastine, mizolastine, mequitazine, mianserine, noberastine, meclizine, norastemizole, olopatadine, picumast, tripelennamine, temelastine, trimeprazine, triprolidine, brompheniramine, chlorpheniramine, dexchlorpheniramine, triprolidine, clemastine, diphenhydramine, diphenylpiraline, tripelennamine, hydroxyzine, methdilazine, promethazine, trimeprazine, azatadine, cyproheptadine, antazoline, pheniramine, pyrilamine, astemizole, terfenadine, loratadine, cetirizine, levocetirizine, fexofenadine, descarboethoxyloratadine, desloratadine, dimenhydrinate, hydroxyzine, and pharmaceutically acceptable salts, esters and derivates thereof.

[0045] The pharmaceutical combination in its base form or its pharmaceutically acceptable salts can be administered by any adequate route, by inhalation or intravenous, although more preferably orally as a solid or liquid.

[0046] Presently the effects that could be caused by the montelukast and ketotifen combination in the treatment of asthma have not been determined. Therefore, the object of this invention is to assess the efficacy of these pharmaceutical agents' combination in an in vitro model of rat tracheal rings constriction.

[0047] This combination improves the therapeutics and offers benefits such as: the administration of the active principles in lesser concentrations than when administered separately; more effectiveness, greater therapeutic power and significantly less probabilities of secondary effects when administered in a combination than when administered separately. Moreover, there is a reduction of secondary effects caused by the separate administration of the compounds, since the doses are lower than those commercially available. Accordingly, the behavior of this combination was proved preclinically, and the interaction and synergism of both compounds were determined along with the optimal proportions of the combination, which had a high degree of efficacy and therapeutic potentiation.

Experimental Model

[0048] Experiment Animals and Operational Description

[0049] Male Wistar rats 300-350 g were sacrificed with a phenobarbital sodium overdose given by intraperitoneal route. The thoracic cavity was exposed, the neck was dissected, and the trachea was removed 1 mm above the tracheal bifurcation and 1 mm below the thyroid cartilage. The sample was submerged in cold Krebs-Henseleit solution bubbled with 95% 02 and 5% CO2, and 3 mm-thick rings were cut. Each ring was placed in an isolated organ chamber, with its bottom anchored by means of a stainless-steel hook and another hook connected to a tension transducer. The chamber was then filled with Krebs-Henseleit solution at 36° C. bubbled with 95% 02 and 5% CO2, pH 7.4±0.5. Two washes were performed, and a 1.5 g load was applied to each ring and were left to rest for 60 minutes. The solution was replaced every 15 minutes and the rings were adjusted until a 1.5 g load was maintained on each ring.

[0050] Once concluded the rest period, the constricting agonists were administered in a cumulative concentration, with a 5 minutes stabilization period for each assay. Once the maximum concentration was achieved, the preparation was washed by replacing three times the solution in the chamber, and the preparation was left to rest for another 15 minutes. Acetylcholine was added to each ring at a concentration of 0.1 mM in the chamber in order to normalize the constricting effect induced by the agonists; the acetylcholine effect accounted for a 100 percent of the tracheal ring constriction.

[0051] Agonist Compounds Used in the Experiments

[0052] The following compounds were employed:

TABLE-US-00001 TABLE 1 Leukotriene D4 final concentrations in the chamber 0.1 nM 0.0496 ng/mL 1 nM 0.4967 ng/mL 10 nM 4.967 ng/mL 100 nM 49.670 ng/mL 1 μM 0.4967 μM 10 μM 4.967 μM

TABLE-US-00002 TABLE 2 Histamine final concentrations in the chamber 1 nM 0.1112 ng/mL 10 nM 1.112 ng/mL 100 nM 11.115 ng/mL 1 μM 111.15 ng/mL 5 μM 555.75 ng/mL 10 μM 1.112 μg/mL 100 μM 11.115 μg/mL

[0053] The antagonist's effects to their own agonists were analyzed using dose-response curves of the agonists in tracheal rings incubated with different antagonists' concentrations. The agonists were added at the end of the resting period, and 5 minutes later the agonists were added in cumulative concentrations. Once reached the maximum concentration, the preparation was washed replacing three times the solution in the chamber, and 15 minutes later acetylcholine was added to each ring at a 0.1 mM concentration in the chamber.

[0054] The analyzed concentrations of the antagonists were as follows:

TABLE-US-00003 Montelukast final concentrations in the chamber 0.003 μM 1.759 ng/mL 0.03 μM 17.59 ng/mL 0.3 μM 175.9 μg/mL 3.0 μM 1.759 μg/mL 30.0 μM 17.59 μg/mL

TABLE-US-00004 Ketotifen final concentrations in the chamber 0.01 μM 3.094 ng/mL 0.1 μM 30.94 ng/mL 1.0 μM 309.43 μg/mL 10.0 μM 3.094 μg/mL 100.0 μM 30.94 μg/mL

[0055] The obtained data were graphed as a percentage of the final concentration compared with that of acetylcholine 0.1 mM, which was 100%.

[0056] The half maximal effective concentration (EC.sub.50) of each dose-response curve of the analyzed agonists was obtained using the sigmoid model, wherein: y=a+(b−a)/(1+10.sup.(logx0-x)*x p)). The antagonists effect percentage curve was constructed with the displacement obtained in the concentration-response curves of an agonist in the presence of an antagonist.

[0057] Using 40 percent of the contraction inhibition effect induced by each agonist, the theoretical and experimental concentrations to be assessed for the 1:1 montelukast and ketotifen combination were calculated. These concentrations were evaluated in isolated rat tracheal rings contracted with the leukotriene D4 and histamine combination at a 1:1 proportion inducing 40 percent of the contraction.

[0058] Data are presented as mean±standard deviation of 2 rings obtained from each of the tracheas of 5 rats. Data were analyzed with the one-way ANOVA test with a Dunett's post-test. EC50's were determined by means of the sigmoid model and the 40 percent of effects was obtained using a linear regression. The t and T values were determined based on a t-values table.

[0059] As a result, the cumulative leukotriene D4 (LTD4) administration induces a concentration-dependent contraction of the rat tracheal rings, this effect has a sigmoid behavior, and the highest concentrations caused a greater contraction than the one induced by acetylcholine 0.1 mM. In this experimental model, the LTD4 EC50 is 42.6 nM. (FIG. 1).

[0060] The cumulative histamine administration induces a concentration-dependent contraction of the rat tracheal rings, this effect has a sigmoid behavior, and this effect has a sigmoid behavior. In this experimental model, the histamine EC50 is 3.54 μM. (FIG. 2).

[0061] The presence of montelukast, a cysLT1 receptors antagonist, induces a concentration-dependent blocking of the contraction induced by the cumulative administration of LTD4. The assayed 30 μM montelukast concentration blocks 75% of the contraction induced by the highest LTD4 concentration. (FIG. 3).

[0062] The presence of ketotifen, a H1 receptors antagonist, induces a concentration-dependent blocking of the contraction induced by the cumulative administration of histamine. The assayed 100 μM ketotifen concentration blocks 65% of the contraction induced by the highest histamine concentration. (FIG. 4).

[0063] FIG. 5 depicts the blocking percentage of the contraction induced by the combined administration of LTD4 and histamine (1:1) causing 40% of the maximum contraction, in the presence of the experimental concentrations of the montelukast and ketotifen combination (1:1).

[0064] Based on these results, an isobologram analysis was performed of the 1:1 montelukast-ketotifen combination blocking the contraction induced by LTD4 and histamine on rat tracheal rings. The obtained experimental data lie under the experimental calculation line, indicating a statistically significant synergic effect of the antagonist combination. (FIG. 6).

[0065] According to this analysis, we can substantiate the fact that the administration of the 1:1 montelukast and ketotifen combination significantly blocks the contraction induced by LTD4 and histamine on rat tracheal rings. Hence, we can determine that this combination could be assessed in models of allergen-induced asthma, and to consider clinical studies of its potential introduction to the treatment of asthma in human beings. The synergic interaction of the montelukast-ketotifen combination in a model of bronchoconstriction induced by a histamine-LTD4 challenge offers several possibilities: its potential use in the treatment of patients with bronchial asthma as a prophylaxis of relapses in the control of asthma. Also, it offers the possibility of reducing the doses or requirements of the active agents and nevertheless achieving the same therapeutic effect, thus reducing the frequency and/or severity of adverse events. This requires corresponding evidences derived from controlled clinical trials of the therapeutic efficacy in asthmatic patients.

[0066] In the current state of the art there are several pharmacological treatments of asthma and other derived diseases; however, there is no other treatment characterized by the combination of the active agents montelukast and ketotifen. The development of this invention provides a real and safe alternative for the control and treatment of asthma, and can reduce treatment durations, therapeutic effects and secondary reactions. The separate administration of these compounds requires approximately 0.01 mg to approximately 1000 mg/day for montelukast and approximately 0.01 mg to approximately 1000 mg/day for ketotifen.

[0067] This invention was developed to be administered by oral, nasal, intramuscular and/or intravenous route, either as immediate release of both drugs, or modified release of one or both drugs, with lower doses, higher therapeutic power and less risk of adverse events.

EXAMPLES

[0068] The following pharmaceutic compositions are described below as non-limiting examples:

Example 1: Oral Administration Composition

[0069]

TABLE-US-00005 Montelukast or pharmaceutically acceptable salt Ketotifen or pharmaceutically acceptable salt Pharmaceutically acceptable excipient and/or carrier and/or additive

Example 2: Intramuscular and Intravenous Administration Composition

[0070]

TABLE-US-00006 Montelukast or pharmaceutically acceptable salt Ketotifen or pharmaceutically acceptable salt Pharmaceutically acceptable excipient and/or carrier and/or additive

Example 3: Nasal Administration Composition

[0071]

TABLE-US-00007 Montelukast or pharmaceutically acceptable salt Ketotifen or pharmaceutically acceptable salt Pharmaceutically acceptable excipient and/or carrier and/or additive

[0072] This invention can be represented in other specific ways without departing from its spirit or essential characteristics, and with any required excipient and/or carrier and/or additive to obtain the desired pharmaceutical form.

[0073] The described embodiments will be in any aspect considered as illustrative and not as limitations. Accordingly, the scope of this invention is defined by the appended claims and not by the above description. Its scope encompasses all modifications within the meaning and range of equivalence of the claims.

[0074] In an integral way, this invention provides the following advantages:

[0075] 1. The combined administration of montelukast and ketotifen in a 1:1 ratio induces a significant blocking of the contraction induced by LTD4 and histamine in rat tracheal rings.

[0076] 2. The potential therapeutic use for patients with atopic diseases such as bronchial asthma, allergic rhinitis and atopic dermatitis, as a prophylaxis of relapses in the control of asthma.

[0077] 3. Offers the possibility of reducing the doses or requirements of the active agents and nevertheless achieving the same therapeutic effect, thus reducing the frequency and/or severity of adverse events.