COMBINATION THERAPY FOR INHALATION ADMINISTRATION

Abstract

The present invention is related to a pharmaceutical composition for nebulization administration comprising an inhaled corticosteroid (ICS), a long-acting β2-agonist (LABA), and optionally a long-acting muscarinic antagonist (LAMA) to be used in the treatment of respiratory diseases, especially in asthma and chronic obstructive pulmonary disease (COPD), and process for the preparation 5 thereof. More particularly, the pharmaceutical compositions herein include beclometasone dipropionate (BPD), formoterol fumarate (FF) and optionally glycopyrronium bromide (GB). The invention also relates to the use of said pharmaceutical formulation in a soft mist inhaler.

Claims

1. A pharmaceutical solution composition for delivery to the pulmonary system via a soft mist inhaler comprising: (a) an inhaled corticosteroid (ICS), (b) a long-acting beta-agonist (LABA), and (c) optionally a long-acting muscarinic antagonist (LAMA), dissolved in a pharmaceutically acceptable solvent, wherein the solvent comprises ethanol in an amount of 75 to 100% v/v.

2-16. (canceled)

17. The pharmaceutical solution composition according to claim 1, wherein said solvent comprises ethanol in an amount of 80-99% v/v.

18. The pharmaceutical solution composition according to claim 1, wherein said solvent comprises ethanol in an amount of 90-97% v/v.

19. The pharmaceutical solution composition according to claim 1, wherein the inhaled corticosteroid is selected from the group consisting of beclomethasone dipropionate, budesonide, ciclesonide, fluticasone propionate; fluticasone furoate, and mometasone; the long-acting beta-agonist is selected from the group consisting of formoterol fumarate, salmeterol, indacaterol, vilanterol, and odolaterol; and the long-acting muscarinic antagonist is selected from the group consisting of glycopyrronium bromide, umeclidinium, aclidinium, ipratropium, tiotropium, and oxitropium.

20. The pharmaceutical solution composition according to claim 19, wherein the inhaled corticosteroid is beclomethasone dipropionate, the long-acting beta-agonist is formoterol fumarate, and the long-acting muscarinic antagonist is glycopyrronium bromide.

21. The pharmaceutical solution composition according to claim 1, wherein the weight ratio by percentage (% w/w) of ICS to LABA is 30-100% to 0.5-50%.

22. The pharmaceutical solution composition according to claim 1, wherein the weight ratio by percentage (% w/w/w) of the ICS, LABA and LAMA is 30-100%, 0.5-50% and 0.5-50%, respectively.

23. The pharmaceutical solution composition according to claim 1, wherein the solution further comprises one or more pharmacologically acceptable acids and/or buffers for adjusting the pH.

24. The pharmaceutical solution composition according to claim 1, wherein said composition does not contain any propellant.

25. The pharmaceutical solution composition according to claim 1, wherein said composition does not contain any preservative.

26. A container comprising a pharmaceutical solution composition according to claim 1, wherein said container is in a form suitable for use with a soft mist inhaler.

27. A kit comprising a container according to claim 26 and a soft mist inhaler.

28. A method of treating or inhibiting an inflammatory and/or obstructive airways disease comprising: administration of the pharmaceutical composition according to claim 1 to a subject having an inflammatory and/or obstructive airways disease.

29. The method according to claim 28, wherein said inflammatory and/or obstructive airways disease is asthma or COPD.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0042] FIG. 1: Aerodynamic size distribution of Formoterol. NGI, Formulation 1 (70% Ethanol) vs Formulation 2 (96% Ethanol).

[0043] FIG. 2: Aerodynamic particle size distribution of A) Beclometasone dipropionate, B) Glycopyrronium bromide and C) Formoterol Fumarate by NGI, Trimbow® pMDI (2 actuations) vs Soft Mist Inhaler (1 actuation).

[0044] FIG. 3: Aerodynamic particle size distribution of A) Beclometasone dipropionate and B) Formoterol Fumarate. NGI, Foster® pMDI (2 actuations) vs Soft Mist Inhaler (1 actuation).

[0045] FIG. 4: Aerodynamic size distribution of Budesonide. NGI, Formulation 1 (70% Ethanol) vs Formulation 2 (96% Ethanol).

[0046] FIG. 5: Aerodynamic size distribution of Formoterol. NGI, Formulation 1 (70% Ethanol) vs Formulation 2 (96% Ethanol).

[0047] FIG. 6: Aerodynamic size distribution of Budesonide. NGI, Symbicort® pMDI (1 actuation) vs Soft Mist Inhaler (1 actuation).

[0048] FIG. 7: Aerodynamic size distribution of Formoterol. NGI, Symbicort® pMDI (1 actuation) vs Soft Mist Inhaler (1 actuation).

DETAILED DESCRIPTION OF THE INVENTION

Definitions

[0049] All terms as used herein in this application, unless otherwise stated, shall be understood in their ordinary meaning as known in the art. Other more specific terms as used in the present application are as set forth below and are intended to apply uniformly throughout the specification and claims unless an otherwise expressly set out definition provides a broader definition.

[0050] Throughout the description and claims the word “comprise” and variations of the word, are not intended to exclude other technical features, additives, components, or steps. Furthermore, the word “comprise” encompasses the case of “consisting of”. The following examples and drawings are provided by way of illustration, and they are not intended to be limiting of the present invention. Furthermore, the present invention covers all possible combinations of particular and preferred embodiments described herein.

[0051] The term “weight ratio by percentage” or “% w/w” refers to the percentage by weight of each active ingredient compared to the total weight of all active ingredients present in the pharmaceutical composition.

[0052] In the context of the present invention, the term “% v/v” refers to the percentage by volume of the solvent used in the formulation.

[0053] When referring to the term “preservative” in the context of the present invention, it refers to a substance effective in inhibiting microbial growth, such as growth of bacteria and fungi, in solutions, such as aqueous solutions. Examples include benzalkonium chloride (BAC) and various forms of edetate (ethylenediaminetetraacetate), such as disodium edetate.

[0054] In the context of the present invention, the term “propellant” refers to substances used to propel the active ingredients in metered dose inhalers, such as pMDIs. Typical propellants are hydrofluorocarbons, such as norflurane.

[0055] The expression “therapeutically effective amount” as used herein, refers to the amount of a compound that, when administered, is sufficient to prevent the development of, or alleviate to some extent, one or more of the symptoms of the disorder or condition being treated. The particular dose of compound administered according to this invention will of course be determined by the particular circumstances surrounding the case, including the particular condition being treated, the duration of the treatment, the nature of any concurrent treatment, and any other factor known to the expert.

[0056] The term “Nominal dose,” as used herein, refers to the loaded dose, which is the amount of active pharmaceutical ingredient (“API”) in an inhalation device prior to administration to the patient. The volume of solution containing the nominal dose is referred to as the “fill volume.”

[0057] The “Fine Particle Fraction” (FPF) is defined as the percentage (%) of particles below 5 μm with respects to the delivered dose. It is also known as the “respirable fraction”. This is defined in European Pharmacopoeia chapter 2.9.18 (Preparations for inhalation: Aerodynamic Assessment of Fine particles) and the equivalent United States Pharmacopoeial chapter is USP <601> Inhalation and Nasal Drug Products: Aerosols, Sprays and Powders—Performance Quality Tests.

[0058] In these chapters, the Fine Particle Dose (FPD) is defined by the mass of active substance less than 5 μm collected when inhalation products are actuated as per Pharmacopoeial chapters through any multistage impactor enabling aerodynamic particle size fractioning of the powder. The “aerodynamic particle size distribution” (APSD) is the product deposition in the multistage impactor obtained from the product actuation. The multiusage apparatus design defined in USP <601> consists of a connector between the impactor and the product (Induction Port), seven stages and a micro-orifice collector (MOC) or Internal Filter holder (IFH) for very fine formulations The Stages consist of removable cups and jets with multiple nozzles particle sizes for all stages except Stage 1, in decreasing order as the powder travels through the impactor. The sizes of these nozzles (μm) determine the aerodynamic deposition of the product powder in each of the stages and therefore enables the FPD calculation previously described.

[0059] The “delivered dose” (DD) is the amount delivered from the inhaler. This term is defined in European Pharmacopoeia chapter “Inhalanda: Preparations for inhalation” and for the USP in the previously mentioned chapter USP<601>. The delivered dose is obtained from the dose collection in a delivered dose apparatus when actuated at standard conditions defined in these chapters.

Solution Formulations

[0060] In one aspect, the present invention concerns a pharmaceutical solution composition for delivery to the pulmonary system via a nebulizer comprising: [0061] (a) an inhaled corticosteroid (ICS), [0062] (b) a long-acting beta-agonist (LABA), and [0063] (c) optionally a long-acting muscarinic antagonist (LAMA), dissolved in a pharmaceutically acceptable solvent, wherein the solvent comprises ethanol in an amount of 75 to 100% v/v. In one embodiment, the solvent comprises ethanol in an amount of 80-99% v/v. In another embodiment, the solvent comprises ethanol in an amount of 85-98% v/v. In a further embodiment, the solvent comprises ethanol in an amount of 90-97% v/v. In still another embodiment, the solvent comprises ethanol in an amount of 92-96% v/v. In yet another embodiment, the solvent comprises ethanol in an amount of 94-96% v/v. In still a further embodiment, the solvent comprises ethanol in an amount of approximately 96% v/v.

[0064] In one embodiment, the pharmaceutical solution composition comprises two active ingredients. In a further embodiment, said two active ingredients are an inhaled corticosteroid, such as beclomethasone dipropionate, and a long-acting beta-agonist, such as formoterol fumarate.

[0065] In one embodiment, the formulations according to the invention contain one or more pharmacologically acceptable acids and/or one or more buffers for adjusting the pH. In another embodiment, said acid is hydrochloric acid. In another embodiment, said buffer is citrate buffer.

[0066] In one embodiment, the formulations according to the invention do not contain a propellant. In a further embodiment, said propellant is a hydrofluorocarbon. In another embodiment, said propellant is norflurane.

[0067] The formulations of the present invention are prepared according to procedures well known in the art that comprise the mixing for some specific time the active ingredients with the solvent, such as ethanol, adjusting of the pH and further mixing for a specific time.

Active Ingredients

[0068] In one embodiment, the inhaled corticosteroid is selected from the group consisting of beclomethasone dipropionate, budesonide, ciclesonide, fluticasone propionate; fluticasone furoate, and mometasone; the long-acting beta-agonist is selected from the group consisting of formoterol fumarate, salmeterol, indacaterol, vilanterol, and odolaterol; and the long-acting muscarinic antagonist is selected from the group consisting of glycopyrronium bromide, umeclidinium, aclidinium, ipratropium, tiotropium, and oxitropium. In a further embodiment, the active substances that may be used in the formulations according to the invention are preferably selected from Beclomethasone dipropionate (BDP), Formoterol fumarate (FF), and optionally Glycopyrronium bromide (GB).

[0069] As used herein, Beclomethasone dipropionate (BDP) is a diester of beclomethasone (also referred to as beclometasone), a synthetic corticosteroid developed for the prophylactic management of mild, moderate, or severe asthma in adults or children and for the prophylactic treatment of chronic reversible obstructive airways disease. The chemical name is 9-chloro-11β,17,21-trihydroxy-16β-methylpregna-1,4-diene-3, 20-dione 17,21-dipropionate.

[0070] Formoterol is a selective β2-adrenergic receptor agonist. It is commercialized as formoterol fumarate (FF) and administered by oral inhalation. Formoterol acts locally in the lung as a bronchodilator. The chemical name of formoterol fumarate dihydrate is N-[2-Hydroxy-5-[(1RS)-1-hydroxy-2-[[(1RS-2-(4-methoxy-phenyl)-1-methylethyl]amino]ethyl]phenyl]formamide (E)-butenedioate dihydrate. Formoterol presents two asymmetric carbons, so four possible isomers exist, forming two racemates. The commercial formoterol is a racemic mixture of R, R (−) and S, S (+) enantiomers which is routinely ensured by a test for specific optical rotation. The content of diastereoisomers R*S* is controlled in the active substance.

[0071] Glycopyrrolate is a long-acting muscarinic antagonist. It is a synthetic quaternary amine known also as glycopyrronium. It is available in oral, intravenous and inhalation forms. Glycopyrrolate is a quaternary ammonium salt with the following chemical name: 3-(2-cyclopentyl-2-hydroxy-2-phenylacetoxy)-1,1-dimethylpyrrolidinium. The molecular formula is C.sub.19H.sub.28NO.sub.3. There are two asymmetric carbons in the molecule. The product is a 50/50% mixture of the enantiomers. Thus, the product is not optically active. The counterion is typically bromide, in which case the long-acting muscarinic antagonist is glycopyrronium bromide (GB).

[0072] In one embodiment, the weight ratio by percentage (% w/w) of beclometasone dipropionate (BDP) to formoterol fumarate (FF) is generally 30-100% to 0.5-50%. In a further embodiment, the weight ratio by percentage is 50-100% to 0.5-25%, respectively. In another embodiment, the weight ratio by percentage is 70-100% to 0.5-10%, respectively. In still another embodiment, the weight ratio by percentage is 80-100% to 1-10%, respectively. In yet another embodiment, the weight ratio by percentage is 90-100% to 2-8%, respectively. In a further embodiment, the weight ratio by percentage is 94% to 6%.

[0073] In one embodiment, the weight ratio by percentage (% w/w/w) of the three active ingredients, beclometasone dipropionate (BDP), formoterol fumarate (FF), and glycopyrronium bromide (GB) is generally 30-100%, 0.5-50% and 0.5-50%, respectively. In a further embodiment the weight ratio by percentage is 50-100%, 0.5-25% and 1-25%, respectively. In another embodiment, the weight ratio by percentage is 70-100%, 0.5-10% and 2-20%, respectively. In still another embodiment, the weight ratio by percentage is 80-100%, 1-10% and 5-17%, respectively. In yet another embodiment, the weight ratio by percentage is 90-100%, 2-8% and 7-15%, respectively. In a further embodiment, the weight ratio by percentage is 84%, 11% and 5%, respectively.

Nebulizers and Soft Mist Inhalers

[0074] The nebulized formulations according to the invention have to meet high quality standards. The formulations according to the invention may be inhaled by oral or nasal route. Those inhalers which are capable of nebulizing a small amount of a liquid formulation in the dosage needed for therapeutic purposes within a few seconds into an aerosol suitable for therapeutic inhalation are particularly suitable. In one embodiment, the nebulizers are those in which an amount of less than 25 microlitres, preferably less than 20 microlitres, most preferably less than 15 microlitres of active substance solution can be nebulized preferably in one puff to form an aerosol having an average particle size (or particle diameter) of less than 10 microns, preferably less than 5 microns, so that the inhalable part of the aerosol already corresponds to the therapeutically effective quantity.

[0075] As used herein, a “nebulized formulation” refers to a solution that is dispersed in the air to form an aerosol. Thus, a nebulized solution is a particular form of an aerosol. A nebulizer is an instrument that is capable of generating very fine liquid droplets for inhalation into the lung. Within this instrument, the nebulizing liquid or solution is atomized into a mist of droplets with a broad size distribution by methods known to those of skill in the art, including, but not limited to, compressed air, ultrasonic waves, or a vibrating orifice. Nebulizers may further contain, e.g., a baffle which, along with the housing of the instrument, selectively removes large droplets from the mist by impact. Soft mist inhalers are a specific type of nebuliser where the mist is produced by high pressure generated during device actuation, such as by releasing a coiled spring. These nebulizers do not need the aid of any external power source. Thus, the mist inhaled into the lung contains fine aerosol droplets. Nebulizers for use herein include, but are not limited to soft mist inhalers.

[0076] In inhalers of this kind, the formulations in solutions are stored in a reservoir. It is important that the active substance formulations used are sufficiently stable when stored and at the same time are such that they can be administered directly, if possible, without any further handling, in accordance with their medical purpose. Moreover, they must not contain any ingredients which might interact with the inhaler in such a way as to damage the inhaler or the pharmaceutical quality of the solution or of the aerosol produced.

[0077] In one embodiment, the nebulizer is a soft mist inhaler. If the formulation according to the invention is nebulized using the technology of a soft mist inhaler, the mass expelled, in at least 97%, preferably at least 98% of one actuation (puff), should correspond to a defined quantity with a range of tolerance of not more than 25%, preferably 20% of this quantity. Preferably, between 5-25 mg, more preferably between 10-15 mg of formulation are delivered as a defined mass per puff.

[0078] Preferably, the medicament combinations according to the invention are used as specified above for preparing a pharmaceutical composition for the treatment of obstructive pulmonary diseases selected from among bronchial asthma, severe asthma, acute asthma attacks, chronic bronchitis and chronic obstructive pulmonary disease (COPD), while it is particularly preferable according to the invention to use them for preparing a medicament for the treatment of bronchial asthma and COPD.

[0079] In view of the above description and the examples below, one of ordinary skill in the art will be able to practice the invention as claimed without undue experimentation. The foregoing will be better understood with reference to the following examples that detail certain procedures for the preparation of formulations according to the present invention.

[0080] The following examples should not be considered exhaustive, but merely illustrative of only a few of the many embodiments contemplated by the present invention.

EXAMPLES

Example 1—Impact of Ethanol in the Solubilization and Aerodynamic Particle Size Distribution

[0081] A study was performed to evaluate the impact of ethanol in the solubilization of the components and the aerodynamic performance of an aerosol solution formulation of beclomethasone dipropionate (BDP), glycopyrronium bromide (GB) and formoterol fumarate (FF).

Formulation

[0082] Beclometasone dipropionate (BDP), glycopyrronium bromide (GLB), formoterol fumarate (FF) and HCl 1N were mixed in 70% ethanol (Formulation 1) and 96% ethanol (Formulation 2) at 25° C. for two hours protected from evaporation. Both formulations were filtrated, and individual assay of all active ingredients were tested by Reverse Phase HPLC.

TABLE-US-00001 TABLE 1 Formulations investigated in this study. Solution API/Excipients Components composition Formulation 1 Beclometasone 87.4 μg Dipropionate Glycopyrronium 10.9 μg Bromide Formoterol 5.2 μg Fumarate HCl 1N 2.5 μg Water for 3.32 μL Irrigation Absolute 7.74 μL Ethanol Formulation 2 Beclometasone 87.4 μg Dipropionate Glycopyrronium 10.9 μg Bromide Formoterol 5.2 μg Fumarate HCl 1N 2.5 μg Ethanol 96% 11.05 μL

HPLC Analytical Methods

[0083] Three independent Reverse Phase HPLC analytical methods were used for the determination of each active ingredient in all the analyses.

[0084] Beclometasone Dipropionate: An Isocratic (1.4 mL/min), HPLC-UV (254 nm) detection was employed in a 60/40 v/v organic (THF:AcN:MeOH 94:434:472 v/v/v)/Aqueous (phosphate buffer pH 2.35) mobile phase elution in a C18 column (Kromasil 100 Å C18; 250×4.6 mm; 5 μm). External Beclometasone Dipropionate standards prepared in the method diluent (56/44 v/v THF:AcN:MeOH 94:434:472 v/v/v/phosphate buffer pH 2.35) were used to quantify the amount of active ingredient present in the sample prepared in the same diluent using the response factor of Standard and Sample solutions. Typical System Suitability Criteria applies as per USP <621> requirements.

[0085] Glycopyrronium Bromide: An Isocratic (1.0 mL/min), HPLC-UV (225 nm) detection was employed with a Mobile Phase composed of Sulphate (heptanosulphonate) buffer pH 5.9/MeOH:AcN:Sulphuric Acid 0.05M in a 61.5:15:23.5:0.3 v/v proportion with a C18 column (Zorbax Extend RR C18, 50×4.6 mm, 3.5 μm). External Glycopirronium Bromide standards prepared in the method diluent (35/65 v/v MeOH/Water) were used to quantify the amount of active ingredient present in the sample prepared in the same diluent using the response factor of Standard and Sample solutions. Typical System Suitability Criteria applies as per USP <621> requirements.

[0086] Formoterol Fumarate: An Isocratic (1.5 mL/min), HPLC-Electrochemical detection was employed with a Mobile Phase composed of 76/24 v/v Phosphate Buffer pH 5.6: AcN v/v with a C18 column (Supelcosil LC-ABZ, 250×4.6 mm, 5 μm). External Formoterol Fumarate standards prepared in the method diluent (100% MeOH) were used to quantify the amount of active ingredient present in the sample prepared in the same diluent using the response factor of Standard and Sample solutions. Typical System Suitability Criteria applies as per USP <621> requirements.

[0087] Data analysis. The Aerodynamic size distribution was plotted as a function of the stage cut-off diameter as the percentage of mass recovered from the induction port filter to the filter.

Aerodynamic Particle Size Distribution (APSD)

[0088] For the APSD evaluation Formulation 1 and 2 were filled into cartridges compatible with a soft mist inhaler (Respimat®). The in vitro aerodynamic particle size distribution of both formulations was evaluated using an impactor, the next generation impactor, NGI (Copley Scientific Ltd) equipped with a mouthpiece adapter for the insertion of the inhaler, an induction port and an internal filter holder (IFH) to capture the smaller particles. The product was tested as per procedure detailed in the European pharmacopoeia chapter 2.9.18 (apparatus E) and US Pharmacopeia chapter <601> (apparatus 6) for Soft Mist Inhalers.

[0089] The NGI was cooled to 5° C. for at least 75 min. The cooling chamber was then opened and, after 30 minutes, the NGI was connected to a HCP5 vacuum pump (Copley Scientific Ltd). For each experiment, ten doses were discharged per NGI actuated at 30 L/min for 5 seconds each.

[0090] After the required actuations, the particles deposited on the different surfaces of the impactor were extracted using a suitable diluent (methanol) and with the help of a Gentle Rocker (Copley Scientific Ltd). Active ingredients were then quantified by HPLC, with the same methods used for the assay testing described above This establishes the distribution of particles according to their aerodynamic particle size.

[0091] Data analysis. The Aerodynamic size distribution was plotted as a function of the stage cut-off diameter as the percentage of mass recovered from the induction port filter to the filter.

Results

[0092] The assay of each active ingredient, summarized in the following Table 2, showed that the formulation providing the best results in terms of higher active ingredient content (in particular beclometasone dipropionate and formoterol fumarate) was Formulation 2—formulation containing 96% ethanol. Without being bound by a particular theory, these results could be ascribed to the poor solubility of these compounds in water.

TABLE-US-00002 TABLE 2 Active ingredient content in Formulation 1 and 2 after filtration. Formulation 1: 70% Ethanol; Formulation 2: 96% Ethanol. Theoretical Active Pharmaceutical concentration Formulation 1 Formulation 2 Ingredient (mg mL.sup.−1) Assay (%).sup.(1) Assay (%).sup.(1) Beclometasone 7.9 40.1 99.7 Dipropionate Glycopyrronium 1.0 99.9 101.0 Bromide Formoterol 0.5 93.8 100.3 Fumarate .sup.(1)Active ingredient content compared to the theoretical amount expressed in percentage.

[0093] Also when comparing Formoterol in formulation 1 & 2, with an equivalent assay, FIG. 1 shows that ethanol has an impact on the aerodynamic particle size distribution. In fact, ethanol allows for the particle size to be modified within the respirable range (extrafine particles or coarser ones).

[0094] The data shows that the higher the ethanol content, the lower is the particle size generated. Extrafine formulations are very desirable as they lead to deeper and more uniform distribution of the inhaled treatment in the lungs.

Conclusions

[0095] In this study, it was demonstrated that beclometasone and formoterol solubilization are highly dependent on ethanol content. Beclometasone and Formoterol were not completely dissolved in 70% ethanol (Formulation 1) while all the active ingredients solubilized in 96% ethanol (Formulation 2).

[0096] In the NGI testing, Formulation 2 showed better aerodynamic properties than Formulation 1.

[0097] These results suggest therefore that a higher ethanol content could lead to more efficient formulations for inhaled therapies.

Example 2—Comparative of Trimbow® Formulation pMDI Versus a Soft Mist Inhaler

[0098] A study was performed to compare the aerodynamic particle size distribution and the emitted dose of a triple combination solution delivered with a soft mist inhaler device (Respimat®) versus the commercialized product Trimbow® pMDI (Triple combination product commercially available). The triple combination consists of the following active ingredients: beclomethasone dipropionate (BDP), glycopyrronium bromide (GB) and formoterol fumarate (FF) at the concentrations shown in Table 3.

[0099] Trimbow® pMDI, a marketed product, was evaluated in this study as a product representative of triple combination products.

Formulation and Inhaler System

[0100] Beclometasone dipropionate (BDP), glycopyrronium bromide (GB), formoterol fumarate (FF) and HCl 1N were mixed in 96% ethanol at 25° C. for two hours. The solution was then filtrated and filled into cartridges compatible with a soft mist inhaler (Respimat®).

[0101] Table 3 summarizes the products investigated in this study.

TABLE-US-00003 TABLE 3 Formulations and devices investigated in this study. Product Formula* Device Name API/Excipients (μg/actuation) Trimbow ® Trimbow ® Beclometasone 87.4 μg pMDI Dipropionate Glycopyrronium 10.9 μg Bromide Formoterol 5.2 μg Fumarate HCl 1N 12.2 μg HFA 56645 μg Ethanol 9.6 μL Soft Mist Triple Beclometasone 87.4 μg Inhaler combination Dipropionate with Soft Glycopyrronium 10.9 μg Mist Bromide Formoterol 5.2 μg Fumarate HCl 1N 2.5 μg Ethanol 96% 11.1 μL *Theoretical mass in μg per delivered dose.

Aerodynamic Particle Size Distribution (APSD)

[0102] The in vitro aerodynamic assessment was carried using a next generation impactor, NGI (Copley Scientific Ltd), equipped with a mouthpiece adapter for the insertion of the inhaler, an induction port and an internal filter holder (IFH) to capture the smaller particles. The product was tested as per procedure detailed in the European pharmacopoeia chapter 2.9.18 (apparatus E) and US Pharmacopeia chapter <601> (apparatus 6) for Soft Mist Inhalers.

[0103] The NGI was cooled to 5° C. for at least 75 min. The cooling chamber was then opened and, after 30 minutes, the NGI was connected to a HCP5 vacuum pump (Copley Scientific Ltd). For each experiment, ten doses were discharged into the NGI at 30 L/min for 5 seconds each.

[0104] After the required actuations, the particles deposited on the different surfaces of the impactor were extracted using a suitable diluent (methanol) and with the help of a Gentle Rocker (Copley Scientific Ltd). Active ingredients were then quantified by HPLC, with the same methods used for the Example 1.

[0105] Data analysis. The in vitro aerodynamic assessment was carried using a next generation impactor, NGI (Copley Scientific Ltd), equipped with a mouthpiece adapter for the insertion of the inhaler, an induction port and an internal filter holder (IFH) to capture the smaller particles. The product was tested as per procedure detailed in the European pharmacopoeia chapter 2.9.18 (apparatus E) and US Pharmacopeia chapter <601> (apparatus 6) for Soft Mist Inhalers.

[0106] The NGI was cooled to 5° C. for at least 75 min. The cooling chamber was then opened and, after 30 minutes, the NGI was connected to a HCP5 vacuum pump (Copley Scientific Ltd). For each experiment, ten doses were discharged into the NGI at 30 L/min for 5 seconds each.

[0107] After the required actuations, the particles deposited on the different surfaces of the impactor were extracted using a suitable diluent (methanol) and with the help of a Gentle Rocker (Copley Scientific Ltd). Active ingredients were then quantified by HPLC, with the same methods used for the Example 1.

[0108] Data analysis. The NGI results were plotted against the stage cut-off diameter as the mass recovered from the induction port to the filter. Fine particle fraction (FPF %), mass median aerodynamic diameter (MMAD), fine particle dose (FPD) were determined from the analysis of the NGI data.

[0109] Results are expressed as the mean value of two NGIs analysis.

Delivered Dose

[0110] The delivered dose was tested according to USP <601> using a dose unit sampling apparatus (DUSA) operating at 28.3 L/min for 4 seconds—the inhalation volume did not exceed 2.0 L. Five doses per device were performed. Active pharmaceutical ingredients (APIs) deposited in the sampling apparatus were quantitatively recovered with a DUSA shaker (Copley Scientific Ltd) and methanol. APIs were, then, quantified by HPLC with the same methods described in Example 1.

[0111] Results are expressed as the average of 5 measurements.

Results

[0112] Generally, drug particles from an inhaler will deposit in different parts of the lungs according to their size; coarser particles will deposit in the mouth and throat, middle size particles will deposit in the primary bronchi central airways while smaller particles will deposit in the terminal bronchi and alveoli. In this study, to determine the delivered dose and the detailed deposition rate of Trimbow® pMDI and a soft mist inhaler, a dose unit sampling apparatus and a next generation impactor (NGI) were used, respectively.

[0113] Table 4 shows that one actuation of both devices released the same dose.

TABLE-US-00004 TABLE 4 Triple combination Beclomethasone, Fomoterol and Glycopyrroniuum in a Soft Mist Inhaler (Respimat ®) and Trimbow ® pMDI. DUSA, Trimbow ® pMDI (1 actuation), Soft Mist Inhaler (1 actuation). Soft Mist Inhaler Trimbow ® pMDI (1 actuation) (1 actuation) API DUSA (μg) RSD* (%) DUSA (μg) RSD (%) Beclometasone 97.1 2.9 98.4 3.2 dipropionate Glycopyrronium 12.3 4.5 11.1 2.8 Bromide Formoterol 6.1 2.8 5.7 2.3 Fumarate *RSD: Relative Standard Deviation.

[0114] However, results from the NGI assay show that one single dose of the soft mist inhaler could simulate the same aerodynamic particle size distribution in terms of respirable particles—particles from stage 1 to internal filter holder (IFH)—as two doses of Trimbow® pMDI. Also, the soft mist inhaler showed lower deposition in the induction port (IP) than Trimbow® pMDI, which means a lower amount of coarser particles that are retained in the mouth and the throat (FIG. 2).

[0115] MMAD values were around 1.2 μm, independently of the device used.

[0116] Taken together, these APSD results show that a soft mist inhaler offers, with 1 actuation containing the same quantity of drug per actuation, the same lung deposition and lower oropharyngeal deposition as Trimbow® pMDI with 2 actuations, that are equivalent to 1 dose, as required per treatment. Thus, translating into a lower number of required actuations to achieve similar therapeutic effects and less drug being delivered to the oropharyngeal region. Due to possible side effects, deposition to the oropharyngeal area is not desired

TABLE-US-00005 TABLE 5 APSD performance of the triple combination Beclomethasone, Fomoterol and Glycopyrronium in a Soft Mist Inhaler (Respimat ®) and the Trimbow ® pMDI. by NGI, Trimbow pMDI (2 actuations) vs Soft Mist Inhaler (1 actuation). Soft Mist Inhaler Trimbow ® pMDI (1 actuation) (2 actuations) MMAD FPF FPD IP MMAD FPF FPD IP API (μm) (%) (μg) (%) (μm) (%) (μg) (%) Bec- 1.2 87.9 91.7 8.3 1.2 42.9 86.0 52.9 lometasone dipropionate Glyco- 1.3 86.7 12.0 9.0 1.2 43.4 10.6 54.2 pyrronium Bromide Formoterol 1.2 87.8 5.3 8.3 1.2 44.6 5.2 53.4 Fumarate

Conclusions

[0117] Soft mist inhalers represent a novel approach to the delivery of inhaled drugs and overcome some of the limitations of DP's and especially pMDIs.

[0118] In this study, though a soft mist inhaler and Trimbow® pMDI showed similar delivered doses, the same lung deposition was observed with the soft mist inhaler with 1 actuation compared to 2 actuations of Trimbow® pMDI—one single actuation with the soft mist inhaler simulates the lung deposition profile (and potentially same therapeutic effect) of two pMDI actuations. It is furthermore shown that similar distributions of particle sizes are achieved for each of the three active ingredients, despite their different physico-chemical properties. Hence, the idea underlying the present invention is applicable to different inhalable drug molecules.

Example 3—Comparative of Foster® pMDI Versus an Equivalent Formulation with a Soft Mist Inhaler (Respimat®)

[0119] A study was performed to compare the aerodynamic particle size distribution of a double combination solution delivered with a soft mist inhaler device versus the commercially available product Foster® pMDI. The double combination consists of the following active ingredients: beclomethasone dipropionate (BDP) and formoterol fumarate (FF) at the concentrations shown in Table 6.

[0120] Foster® 100/6 μg pMDI, a marketed product, was evaluated in this study as product representative of double combination products.

Formulation and Inhaler System

[0121] Beclometasone dipropionate (BDP), formoterol fumarate (FF) and HCl 1N were mixed in 96% ethanol at 25° C. for two hours. The solution was then filtrated and filled into cartridges compatible with a Soft Mist Inhaler.

[0122] Table 6 summarizes the products investigated in this study.

TABLE-US-00006 TABLE 6 Formulations and devices investigated in this study. Formula* Device Product Name API/Excipients (μg/actuation) Foster ® Foster ® Beclometasone 84.6 μg pMDI Dipropionate Formoterol 5.0 μg Fumarate HCl 1N 11.9 μg HFA 55034 μg Ethanol 9.3 μL Soft Mist Double Beclometasone 84.6 μg Inhaler combination Dipropionate in Soft Mist Formoterol 5.0 μg Inhaler Fumarate HCl 1N 2 μg Etanol 96% 11.1 μL *Theoretical mass in μg per delivered dose.

Aerodynamic Particle Size Distribution (APSD)

[0123] The in vitro aerodynamic assessment was carried using a next generation impactor, NGI (Copley Scientific Ltd), equipped with a mouthpiece adapter for the insertion of the inhaler, an induction port and an internal filter holder (IFH) to capture the smaller particles. The product was tested as per procedure detailed in the European and US Pharmacopeia for Soft Mist Inhalers and pMDIs.

[0124] The NGI was cooled to 5° C. for at least 75 min. The cooling chamber was then opened and, after 30 minutes, the NGI was connected to a HCP5 vacuum pump (Copley Scientific Ltd). For each experiment, ten doses were discharged into the NGI at 30 L/min for 5 seconds each.

[0125] After the required actuations, the particles deposited on the different surfaces of the impactor were extracted using a suitable diluent (methanol) and with the help of a Gentle Rocker (Copley Scientific Ltd). Active ingredients were then quantified by HPLC, with the same methods used for the Example 1.

[0126] Data analysis. The in vitro aerodynamic assessment was carried using a next generation impactor, NGI (Copley Scientific Ltd), equipped with a mouthpiece adapter for the insertion of the inhaler, an induction port and an internal filter holder (IFH) to capture the smaller particles. The product was tested as per procedure detailed in the European pharmacopoeia chapter 2.9.18 (apparatus E) and US Pharmacopeia chapter <601> (apparatus 6) for Soft Mist Inhalers.

[0127] The NGI was cooled to 5° C. for at least 75 min. The cooling chamber was then opened and, after 30 minutes, the NGI was connected to a HCP5 vacuum pump (Copley Scientific Ltd). For each experiment, ten doses were discharged into the NGI at 30 L/min for 5 seconds each.

[0128] After the required actuations, the particles deposited on the different surfaces of the impactor were extracted using a suitable diluent (methanol) and with the help of a Gentle Rocker (Copley Scientific Ltd). Active ingredients were then quantified by HPLC, with the same methods used for the Example 1.

[0129] Data analysis. The NGI was plotted against the stage cut-off diameter as the mass recovered from the induction port to the filter. Fine particle fraction (FPF %) and mass median aerodynamic diameter (MMAD) were determined from the analysis of the NGI data.

[0130] Results are expressed as the mean value of two NGIs analysis.

Results

[0131] The deposition rate of Foster® pMDI and Soft Mist Inhaler was determined by a next generation impactor. Results showed that one single dose of Soft Mist Inhaler double the FPF values of one actuation of Foster® pMDI. MMAD values were around 1.2 μm, independently the device used (Table 7 and FIG. 3).

TABLE-US-00007 TABLE 7 APSD performance of the Soft Mist Inhaler and the Foster ® pMDI. NGI, Foster pMDI (2 actuation) vs Soft Mist Inhaler (1 actuation). Soft Mist Inhaler Trimbow ® pMDI (1 actuation) (2 actuations) AP MMAD (μm) FPF (%) MMAD (μm) FPF (%) Beclometasone 1.1 91.3 1.2 49.4 dipropionate Formoterol 1.2 90.4 1.2 45.8 Fumarate

Conclusions

[0132] Results from this study show that one single actuation with Soft Mist Inhaler could simulate the lung deposition rate (and potentially same therapeutic effect) of two pMDI actuations.

[0133] Taken together, these APSD results were in agreement with those obtained in example 2 and show the potential of Soft Mist Inhaler not only for triple combination therapies but also for double combination treatments.

Example 4—Impact of Ethanol in the Aerodynamic Particle Size Distribution of a Double Combination Therapy

[0134] A study was performed to evaluate the impact of ethanol in the aerodynamic particle size distribution of a double combination solution delivered with a soft mist inhaler device (Respimat®). The double combination consists of the following active ingredients: budesonide (BU) and formoterol fumarate (FF) at the concentrations shown in Table 8.

Formulation and Inhaler System

[0135] Budesonide (BU) and formoterol fumarate (FF) were mixed in 70% ethanol (Formulation 1) and 96% ethanol (Formulation 2) at 25° C. for two hours protected from evaporation. Both formulations were then filtrated, and filled into cartridges compatible with soft mist inhaler (Respimat®).

[0136] Table 8 summarizes the products investigated in this study.

TABLE-US-00008 TABLE 8 Formulations investigated in this study. Formula* Solution API/Excipients (μg/actuation) Formulation 1 Budesonide 160.0 μg Formoterol 4.5 μg Fumarate Water for 3.32 μL Irrigation Ethanol 7.74 μL absolute Formulation 2 Budesonide 160.0 μg Formoterol 4.5 μg Fumarate Ethanol 96% 11.05 μL *Theoretical mass in μg per delivered dose.

HPLC Analytical Methods

[0137] One Reverse Phase HPLC-UV analytical method was used for the determination of both active ingredients.

[0138] An Isocratic (1.0 mL/min), HPLC-UV detection (Budesonide wavelength: 240 nm, Formoterol Fumarate wavelength: 214 nm) was employed with a Mobile Phase composed of 50/50 v/v ACN/0.1% Formic Acid with a C18 column (Hypersil BDS, 250×4.6 mm, 5 μm). External Formoterol Fumarate and Budesonide standards prepared in the method diluent (75/25 v/v MeOH/water) were used to quantify the amount of active ingredient present in the sample prepared in the same diluent using the response factor of Standard and Sample solutions. Typical System Suitability Criteria applies as per USP <621> requirements.

Aerodynamic Particle Size Distribution (APSD)

[0139] For the APSD evaluation Formulation 1 and 2 were filled into cartridges compatible with a soft mist inhaler (Respimat®). The in vitro aerodynamic particle size distribution of both formulations was evaluated using an impactor, the next generation impactor, NGI (Copley Scientific Ltd) equipped with a mouthpiece adapter for the insertion of the inhaler, an induction port and an internal filter holder (IFH) to capture the smaller particles. The product was tested as per procedure detailed in the European pharmacopoeia chapter 2.9.18 (apparatus E) and US Pharmacopeia chapter <601> (apparatus 6) for Soft Mist Inhalers.

[0140] The NGI was cooled to 5° C. for at least 75 min. The cooling chamber was then opened and, after 30 minutes, the NGI was connected to a HCP5 vacuum pump (Copley Scientific Ltd). For each experiment, ten doses were discharged per NGI actuated at 28.3 L/min for 5 seconds each.

[0141] After the required actuations, the particles deposited on the different surfaces of the impactor were extracted using a suitable diluent (75/25 v/v MeOH/water) and with the help of a Gentle Rocker (Copley Scientific Ltd). Active ingredients were then quantified by HPLC as described above.

[0142] Data analysis. The NGI results were plotted against the stage cut-off diameter as the mass recovered from the induction port to the filter. Fine particle fraction (FPF %), mass median aerodynamic diameter (MMAD), fine particle dose (FPD) were determined from the analysis of the NGI data.

[0143] Results are expressed as the mean value of two NGIs analysis.

Results

[0144] The deposition rate of Formulation 1 and Formulation 2 was determined by a next generation impactor. Results showed that ethanol has an impact on the aerodynamic particle size distribution (Table 9 and FIGS. 4 and 5). In fact, a higher ethanol content led to an increase in FPF values and a lower particle size distribution. Extrafine particle size distribution are very desirable as they lead to more efficient treatments in inhaled therapies.

TABLE-US-00009 TABLE 9 APSD performance of Formulation 1 and Formulation 2 delivered with a Soft Mist Inhaler. Formulation 1 Formulation 2 (70% EtOH) (96% EtOH) MMAD FPF FPD MMAD FPF FPF API (μm) (%) (μg) (μm) (%) (μg) Budesonide 2.6 68.5 119.1 1.1 89.2 141.0 Formoterol 2.7 68.4 3.3 1.1 89.7 3.7 Fumarate

Conclusions

[0145] In inhalation therapies, extrafine formulations lead to deeper and more uniform lung distribution. In this study, it was demonstrated that a higher ethanol content provides a lower particle size distribution and therefore gives better aerodynamic properties to the formulation. Results are in line with Example 1 and confirm the potential that ethanol has to generate more efficient formulations for inhaled therapies.

Example 5—Comparative of Symbicort® pMDI Versus an Equivalent Ethanolic Formulation Delivered with a Soft Mist Inhaler (Respimat®)

[0146] A study was performed to compare the aerodynamic particle distribution of a double combination ethanolic solution delivered with a soft mist inhaler device (Respimat®) versus the commercialized product Symbicort® pMDI (double combination product commercially available). The double combination consists of the following active ingredients: Budesonide (BU) and formoterol fumarate (FF) at the concentrations shown in Table 10.

[0147] Symbicort® 160/4.5 μg pMDI, a marketed product, was evaluated in this study as product representative of double combination products with no ethanol in their formula.

Formulation and Inhaler System

[0148] Budesonide (BU) and formoterol fumarate (FF) were mixed in 96% ethanol at 25° C. for two hours protected from evaporation. The solution was then filtrated and filled into cartridges compatible with soft mist inhaler (Respimat®).

[0149] Table 10 summarizes the products investigated in this study.

TABLE-US-00010 TABLE 10 Formulations investigated in this study. Formula* Device API/Excipients (μg/actuation) Symbicort ® Budesonide 160.0 μg pMDI Formoterol 4.5 μg Fumarate HFA227 q.s. Povidone q.s. Macrogol q.s. Soft mist Budesonide 160.0 μg inhaler Formoterol 4.5 μg Fumarate Ethanol 96% 11.05 μL *Theoretical mass in μg per delivered dose.

Aerodynamic Particle Size Distribution (APSD)

[0150] The in vitro aerodynamic particle size distribution of both formulations was evaluated using an impactor, the next generation impactor, NGI (Copley Scientific Ltd) equipped with a mouthpiece adapter for the insertion of the inhaler, an induction port and an internal filter holder (IFH) to capture the smaller particles. The product was tested as per procedure detailed in the European and US Pharmacopoeia for Soft Mist Inhalers and pMDIs.

[0151] The NGI was cooled to 5° C. for at least 75 min. The cooling chamber was then opened and, after 30 minutes, the NGI was connected to a HCP5 vacuum pump (Copley Scientific Ltd). For each experiment, ten doses were discharged per NGI actuated at 28.3 L/min for 5 seconds each.

[0152] After the required actuations, the particles deposited on the different surfaces of the impactor were extracted using a suitable diluent (75/25 v/v MeOH/water) and with the help of a Gentle Rocker (Copley Scientific Ltd). Active ingredients were then quantified by HPLC, with the same methods described in example 4.

[0153] Data analysis. The NGI was plotted against the stage cut-off diameter as the mass recovered from the induction port to the filter. Fine particle fraction (FPF %), and mass median aerodynamic diameter (MMAD) were determined from the analysis of the NGI data.

[0154] Results are expressed as the mean value of two NGIs analysis.

[0155] Results

[0156] The deposition rate of Symbicort® pMDI and Soft Mist Inhaler was determined by a next generation impactor. Results showed that the Soft Mist Inhaler composition has an impact on the aerodynamic particle size distribution (Table 11 and FIGS. 6 and 7). The Soft Mist Inhaler composition led to a smaller particle size distribution achieving MMAD values of around 1.1 μm. This is very positive since small particle size distribution leads to deeper and more uniform lung distribution.

[0157] Moreover, ethanolic solutions delivered with a soft mist inhaler led FPF values of around 1.5-fold higher than the pMDI product.

TABLE-US-00011 TABLE 11 APSD performance of Symbicort ® pMDI and the Soft Mist Inhaler. NGI, Symbicort ® pMDI (1 actuation) vs Soft Mist Inhaler (1 actuation). Symbicort ® pMDI Soft Mist Inhaler (1 actuation) (1 actuation) API MMAD (μm) FPF (%) MMAD (μm) FPF (%) Budesonide 3.7 49.7 1.1 89.2 Formoterol 3.3 58.7 1.1 89.7 Fumarate

Conclusions

[0158] In this study, it was demonstrated that the Soft Mist Inhaler composition leads to smaller particle size distributions. Thus, translating into a deeper and more efficient lung distribution. Moreover, the solution delivered with the Soft Mist Inhaler showed higher FPF values, resulting in lower doses required to achieve similar therapeutic effects than the pMDI and potentially lower side effects due to less drug being delivered to the oropharyngeal region.