BR112016026460B1 - MICROPARTICLES OF MULTIPLE COMPONENTS, THEIR PREPARATION PROCESS, PHARMACEUTICAL FORMULATIONS AND INHALERS - Google Patents

Abstract

MICROPARTICLES OF MULTIPLE COMPONENTS, THEIR PREPARATION PROCESS, PHARMACEUTICAL FORMULATIONS AND INHALERS for the prevention and/or treatment of respiratory diseases.

Description

TECHNICAL FIELD

[001] The present invention relates to particles comprising three active ingredients for administration by means of inhalation.

[002] In particular, the invention relates to particles comprising a combination of an anticholinergic, a beta2-adrenoceptor agonist, and an inhaled corticosteroid, a process for their preparation and their use for the prevention and/or treatment of respiratory diseases .

BACKGROUND OF THE INVENTION

[003] Respiratory diseases are a common and important cause of illness and death around the world. In reality, many people are affected by obstructive and/or inflammatory lung disease, a category characterized by easily flexible and inflamed airways, obstruction to airflow, problems with exhalation, and frequent hospitalizations and clinic visits. Types of obstructive and/or inflammatory lung disease include asthma, bronchiectasis, bronchitis, and chronic obstructive pulmonary disease (COPD).

[004] In particular, chronic obstructive pulmonary disease (COPD) is a multicomponent disease characterized by airflow limitation and airway inflammation. COPD irritations have a considerable impact on patients' quality of life, daily activities and general well-being and are a major affliction in the healthcare system. Thus, the goals of controlling COPD include not only alleviating symptoms and preventing disease progression, but also preventing and treating irritations.

[005] While available therapies improve clinical symptoms and decrease airway inflammation, they do not unequivocally reduce long-term progression or address all components of the disease. With the burden of COPD continuing to increase, research into new and improved treatment strategies to optimize pharmacotherapy is progressive, and in particular, combination therapies, with a view to their complementary modes of action allowing multiple components of the disease to be treated. . Evidence from recent clinical trials indicates that triple therapy, combining an anticholinergic with an inhaled corticosteroid, and a long-acting β2-adrenoceptor agonist, may provide clinical benefits additional to those associated with each treatment alone in patients with more severe COPD.

[006] Currently, there are several recommended classes of therapy for COPD, of which bronchodilators such as β2-agonists and anticholinergics are the mainstay of symptom control in mild and moderate disease, prescribed on an as-needed basis to mild COPD and as a maintenance therapy for moderate COPD.

[007] Such bronchodilators are efficiently administered by means of inhalation, thereby increasing the therapeutic index and reducing the side effects of the active material.

[008] For the treatment of more severe COPD, guidelines recommend the addition of inhaled corticosteroids (ICSs) to long-acting bronchodilator therapy. Combination therapies have been investigated with a view to their complementary modes of action allowing multiple components of the disease to be treated. Data from recent clinical trials indicate that triple therapy, combining an anticholinergic with an ICS and a long-acting β2-agonist (LABA), may provide clinical benefits additional to those associated with each treatment alone in patients with moderate to severe COPD.

[009] Furthermore, there is evidence suggesting synergistic actions of LABA and ICS since both active ingredients are present at the same site of action, for example, the small peripheral airways of the pulmonary tree. Without being bound by theory, this could also occur if the antimuscarinic drug is released at the said site of action.

[0010] An interesting triple combination, currently under investigation, includes:

[0011] i) formoterol, particularly its fumarate dihydrate salt (hereinafter referred to as FF), a long-acting beta-2 adrenergic receptor agonist currently used clinically in the treatment of bronchial asthma, COPD and related diseases;

[0012] ii) rac-glycopyrronium bromide (hereinafter referred to as GB), an antimuscarinic drug recently approved for the maintenance treatment of COPD;

[0013] iii) beclomethasone dipropionate (hereinafter referred to as BDP) a potent anti-inflammatory steroid or corticosteroid, available under a large number of trademarks for the prophylaxis and/or treatment of asthma and other respiratory disorders.

[0014] On the other hand, current combination inhalation products can be subject to great variability in the release of the dose of each active ingredient, which in turn can be perpetuated as a function of the storage conditions of the product.

[0015] Consequently, it would be advantageous to provide a process enabling the preparation of combination particles for inhalation that will allow all three active ingredients to be released without significant dose variations.

[0016] In addition, there is still a need for improved therapeutic control of individuals with respiratory diseases affecting the small peripheral airways.

[0017] For this reason it would be highly advantageous to provide a process allowing the preparation of combination particles according to which all three active ingredients could simultaneously reach the distal tract of the respiratory tree and consequently improving the results of the small airways and associated control.

[0018] In the prior art, different methodologies have been proposed for the preparation of particles incorporating a combination of two or more active ingredients. For example, WO 02/28377, WO 2010/097188, and WO 2013/021199 describe particles incorporating, inter alia, a LABA and an ICS.

[0019] In any case, none of the aforementioned documents describe particles incorporating a combination of formoterol salts, beclomethasone dipropionate and glycopyrronium salts. Also, everyone is silent about the problem of reaching the distal tract of the respiratory tree.

SUMMARY OF THE INVENTION

[0020] The invention is directed to multicomponent microparticles for use in an inhalation formulation, each microparticle comprising a combination of beclomethasone dipropionate, a pharmaceutically acceptable salt of formoterol, and a pharmaceutically acceptable salt of glycopyrronium in a ratio comprised between 35: 10: 55 and 94: 1: 5 weight/weight/weight, according to which said microparticles are characterized by means of a formatting factor comprised between 0.8 and 1.15, preferably between 0.9 and 1.10, more preferably between 0.95 and 1.05.

[0021] Advantageously at least 90% of all of the above microparticles having a volume diameter equal to or less than 4.5 microns, preferably equal to or less than 4.0 microns, and the average diameter of the volume of said microparticles is comprised between 1.0 and 3.0 microns, preferably 1.2 and 2.5 microns, more preferably between 1.5 and 2.2 microns.

[0022] In a second aspect, the invention provides pharmaceutical aerosol formulations for pressurized metered dose inhalers (pMDIs) comprising the above microparticles in suspension in a liquefied propellant gas.

[0023] In a third aspect, the invention provides a pressurized metered dose inhaler (pMDI) comprising a canister filled with the aforementioned pharmaceutical aerosol formulation, and a dosimeter valve for delivering a therapeutically effective dose per day of the active ingredient.

[0024] In a fourth aspect, the invention relates to a dry powder pharmaceutical formulation comprising the above microparticles and optionally a carrier.

[0025] In a fifth aspect, the invention provides a dry powder inhaler filled with the aforementioned dry powder formulation.

[0026] In a sixth aspect, the invention is directed to a process for preparing the claimed microparticles, the process comprising the steps of:

[0027] a) preparing a solution of the three active ingredients in a predetermined ratio in a suitable solvent;

[0028] b) generating an aerosol from the solution of said three active ingredients;

[0029] c) drying the atomized droplets to produce the microparticles; It is

[0030] d) isolating the microparticles produced.

[0031] In a seventh aspect, the invention relates to the claimed microparticles for use in the prevention and/or treatment of an inflammatory or obstructive airways disease, such as asthma or chronic obstructive pulmonary disease (COPD).

[0032] In an eighth aspect, the invention relates to a method of preventing and/or treating an inflammatory or obstructive airways disease, such as asthma or chronic obstructive pulmonary disease (COPD), which comprises administration by means for inhaling an effective amount of the microparticles of the invention.

[0033] In a ninth aspect, the invention relates to the use of the claimed microparticles in the production of a medicine for the prevention and/or treatment of an inflammatory or obstructive disease of the airways, such as asthma or obstructive pulmonary disease chronic disease (COPD).

[0034] In a tenth aspect, the invention relates to multicomponent microparticles for use in an inhalation formulation comprising a combination of beclomethasone dipropionate, a pharmaceutically acceptable salt of formoterol, and a pharmaceutically acceptable salt of glycopyrronium in a ratio comprised between 35: 10: 55 and 94: 1: 5 weight/weight/weight, according to which said microparticles are characterized by means of a formatting factor comprised between 0.95 and 1.05 and are obtainable through a process comprising the steps of:

[0035] a) preparing a solution of the three active ingredients in a predetermined ratio in a suitable solvent;

[0036] b) generate an aerosol from the solution of said three active ingredients and

[0037] c) drying the atomized droplets to produce the microparticles.

DEFINITIONS

[0038] The terms "muscarinic receptor antagonists", "antimuscarinic drugs" and "anticholinergic drugs" may be used interchangeably.

[0039] The term "pharmaceutically acceptable glycopyrronium salt" refers to a salt of the compound 3-[(cyclopentylhydroxyphenylacetyl)oxy]-1,1-dimethylpyrrolidinium.

[0040] The term "pharmaceutically acceptable salt of formoterol" refers to a salt of the compound 2'-hydroxy-5'-[(RS)-1-hydroxy-2-{[(RS)-p-methoxy-a- methylphenethyl]amino}ethyl]formanilide.

[0041] The term "beclomethasone dipropionate" refers to the compound (8S,9R,10S,11S,13S,14S,16S,17R)-9-chloro-11-hydroxy-10, 13,16-trimethyl-3- oxo-17-[2-(propionyloxy)acetyl]-6,7,8,9,10,11,12,13,14, 15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl propionate.

[0042] The term "pharmaceutically acceptable salt" comprises organic and inorganic salts. Examples of organic salts may include formate, acetate, trifluoroacetate, propionate, butyrate, lactate, citrate, tartrate, malate, maleate, succinate, methanesulfonate, benzenesulfonate, xinafoate, pamoate, and benzoate. Examples of inorganic salts may include fluoride, bromide, iodide, phosphate, nitrate and sulfate chloride.

[0043] The term "solvent" is used to mean the medium in which the active ingredients are dissolved, while the term "antisolvent" is used to mean the medium in which crystallization takes place.

[0044] The term "multi-component particle" refers to the smallest discrete single particle comprising a combination of three active ingredients. Said single particle is designed in spherical shape.

[0045] The term "micronized" refers to a substance having a size of a few microns.

[0046] The term "coarse" refers to a substance having a size of one or a few hundred microns.

[0047] In general terms, the particle size of particles is quantified by measuring a characteristic equivalent sphere diameter, known as volume diameter, by means of laser diffraction.

[0048] The particle size can also be quantified by measuring the mass diameter by means of a known suitable instrument, such as, for example, the sieve analyzer.

[0049] The volume diameter (VD) is related to the mass diameter (MD) by the density of the particles (assuming size-independent density for the particles).

[0050] In the present application, the particle size of the active ingredients and the fraction of the fine particles is expressed in terms of volume diameter, while that of the coarse particles is expressed in terms of mass diameter.

[0051] The particles have a normal (Gaussian) distribution which is defined in terms of the mass or volume average diameter (VMD or MMD) which corresponds to the mass or volume diameter of 50 percent by weight of the particles, and optionally in terms of mass or volume diameter of 10% and 90% of the particles, respectively.

[0052] Another common methodology for defining the particle size distribution is to quote three values: i) the average diameter d (0.5) which is the diameter in which 50% of the distribution is above and 50% is below; ii) d (0.9), where 90% of the distribution is below this value; iii) d (0.1), where 10% of the distribution is below this value.

[0053] The coverage is the width of the distribution based on the 10%, 50% and 90% quantile and is calculated according to the formula.

[0054] In general terms, particles having the same or a similar VMD or MMD may have a different particle size distribution, and in particular a different width of the Gaussian distribution as represented by the values d(0.1) and d (0.9).

[0055] In aerosolization, the particle size is expressed as mass aerodynamic diameter (MAD), while the particle size distribution is expressed in terms of mass median aerodynamic diameter (MMAD) and Geometric Standard Deviation ( GSD). MAD indicates the ability of particles to be transported suspended in an air stream. The MMAD corresponds to the 50 weight percent mass aerodynamic diameter of the particles.

[0056] In the final formulation, the particle size of the microparticles of the invention can be determined by means of scanning electron microscopy according to methods known to the person skilled in the art.

[0057] The term 'synergistic' means that the activity of the active ingredients is more than would be expected by adding up their respective individual activities in a given assay.

[0058] The term 'ordered or interactive mixture' refers to a powder formulation for inhalation comprising a physiologically acceptable, pharmacologically inert carrier substance, to which the particles of the micronized active compound are bonded by means of adhesion in order to, in this way, obtaining and maintaining adequate mixed material, i.e. homogeneity of the mixture.

[0059] The term 'relatively highly fissured surface' means a surface on which there are cracks and depressions and other recessed regions, collectively referred to herein as fissures. Such surface of coarse excipient particles can be defined in terms of roughness coefficients or cracking index as described in WO 01/78695 and WO 01/78693 and can be characterized according to the Description reported here.

[0060] The term 'hard pellets' refers to spherical or semi-spherical units whose core is produced from coarse excipient particles.

[0061] The term 'good flowability' refers to a formulation that is easy to handle during the production process and is capable of ensuring an accurate and reproducible release of the therapeutically effective dose.

[0062] The flow characteristics can be evaluated by different tests, such as, friction angle, Carr index, Hausner ratio or flow rate through an orifice.

[0063] In the context of the present application the flow properties were tested by measuring the flow rate through an orifice according to the method described in European Pharmacopoeia (Eur. Ph.) 7.3, 7th Edition.

[0064] The expression 'good homogeneity' refers to a powder in which, during mixing, the uniformity of distribution of a component, expressed as coefficient of variation (CV) also known as relative standard deviation (RSD), is less than than 5.0%. It is generally determined according to known methods, for example by taking samples of different parts of the powder and testing the component by means of HPLC or other equivalent analytical methods such as UPLC.

[0065] The expression 'respirable fraction' refers to an index of the percentage of active ingredient particles that would reach the lungs in a patient.

[0066] The respirable fraction is evaluated using a suitable in vitro mechanism, such as the Andersen Cascade Impactor (ACI), Multi Stage Liquid Impinger (MLSI) or Next Generation Impactor (NGI), according to the procedures reported in the common Pharmacopoeias, in particular in the European Pharmacopoeia (Eur. Ph.) 7.3, 7th Edition. It is calculated by the ratio of the percentage of fine particle mass (formerly fine particle dose) to the delivered dose.

[0067] The released dose is calculated from the cumulative deposition on the mechanism, while the fine particle mass is calculated from the deposition of particles having a diameter of < 5.0 microns.

[0068] In the context of the present application, the formulation is defined as an extra-fine formulation when it is capable of releasing a fraction of the particles having a particle size equal to or less than 2.0 microns equal to or greater than 20%, of preferably equal to or greater than 25%, more preferably equal to or greater than 30% and/or is capable of releasing a fraction of the particles having a particle size equal to or less than 1.0 micron equal to or greater than 10%, preferably equal to or greater than 20%.

[0069] The expression 'chemically stable' refers to an active ingredient that, in the storage of microparticles, satisfies the requirements of the EMEA Guideline CPMP/QWP/122/02 referring to 'Stability Testing of Existing Active Substances and Related Finished Products '.

[0070] The expression 'physically stable' refers to microparticles that show substantially no change in morphology, no transition from amorphous to crystalline state or vice versa, no growth in particle size during storage for at least one month in ambient temperature and 60% relative humidity, as determined according to methods known to the person skilled in the art.

[0071] The expression 'good constancy of the ratio of active ingredients' means that the three active ingredients, after the release of a single therapeutic dose, maintain substantially the same ratio as the predetermined ratio of said two active ingredients in the formulation, i.e. that of the relative standard deviation (RSD) of the ratio of drug amounts measured in an in vitro mechanism, such as the NGI is less than 15%, preferably less than 10%.

[0072] The term 'prevention' means a methodology to reduce the risk of onset of a disease.

[0073] The term 'treatment means a methodology for obtaining benefits or desired results, including clinical outcomes. The desired clinical benefit or outcomes may include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, decreasing the extent of disease, stabilizing (i.e., not getting worse) state of disease, preventing further spread of disease, delay or decrease in disease progression, improvement or palliation of the disease state, and remission (either partial or complete), either detectable or undetectable. The term can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0074] According to the Global Initiative for Asthma (GINA), 'severe persistent asthma' is defined as a form characterized by symptoms per day, frequent irritations, frequent nocturnal asthma symptoms, limitation of physical activities, forced expiratory volume in one second (FEV1) equal to or less than 60% predicted and with a variability greater than 30%.

[0075] According to the guidelines of the Global initiative for chronic Obstructive Pulmonary Disease (GOLD), 'severe COPD' is a form characterized by a ratio between FEV1 and Forced Vital Capacity (FVC) less than 0.7 and FEV1 between 30% and 50% of predicted. The most severe form is also characterized by chronic respiratory failure.

[0076] The expression 'single therapeutically effective dose' means the amount of active ingredient administered at one time by means of inhalation in actuation of the inhaler. Said dose may be delivered in one or more actuations, preferably one actuation (injection) of the inhaler.

[0077] 'Actuation' refers to the release of the active ingredients from the device through a single activation (eg, mechanical or respiratory).

[0078] The expression 'water insoluble or poorly water soluble' is used with reference to water solubility as defined in the European Pharmacopoeia 4th Edition, 2003, page 2891.

[0079] The term 'UPLC-PDA' refers to an Ultra Performance Liquid Chromatography instrument coupled with a Photodiode Array detector.

FIGURE

[0080] Figure 1 - The SEM micrographs of the microparticles of the invention obtained through spray drying.

DETAILED DESCRIPTION OF THE INVENTION

[0081] The invention is directed to multicomponent microparticles for use in an inhalation formulation, each microparticle comprising a combination of beclomethasone dipropionate, a pharmaceutically acceptable salt of formoterol, and a pharmaceutically acceptable salt of glycopyrronium.

[0082] Formoterol may be present in the form of any pharmaceutically acceptable salts and/or its solvate form, preferably in the form of fumarate dihydrate salt.

[0083] Glycopyrronium may be employed in the form of any of the pure diastereoisomers or enantiomers or any combination thereof in the practice of the present invention. In a preferred embodiment, the (3S,2'R), (3R,2'S) 1:1 racemic mixture is employed, also known as rac-glycopyrronium.

[0084] Said active ingredient may be present in the form of any pharmaceutically acceptable salts and/or its solvate form, preferably in the form of bromide or chloride monohydrate, more preferably in the form of bromide salt.

[0085] Beclomethasone dipropionate can be anhydrous or present in the form of monohydrate.

[0086] In a preferred embodiment of the invention, each microparticle consists of a combination of formoterol fumarate or its dihydrate form thereof, glycopyrronium bromide and beclomethasone dipropionate.

[0087] The weight ratio in which the three active ingredients, for example, beclomethasone dipropionate, a pharmaceutically acceptable salt of formoterol, and a pharmaceutically acceptable salt of glycopyrronium, are present in the microparticles is predetermined in such a way as to delivering the desired single therapeutically effective dose of each active ingredient.

[0088] As follows, the weight ratio between the three active ingredients is given by producing the reference to the anhydrous form of beclomethasone dipropionate (BDP), to the formoterol fumarate dihydrate salt (FF), and to the glycopyrronium bromide (GB) salt.

[0089] Advantageously, said ratio could be comprised between 35: 10: 55 and 94: 1: 5 weight/weight/weight. In a preferred embodiment the ratio could be comprised between 70:10:20 to 92:2:6.

[0090] Examples of ratios according to the invention are: 84.4: 5.1: 10.5 weight/ weight/ weight to release 100 microg of BDP, 6 microg of FF, and 12.5 microg of GB ; 91.5: 2.7: 5.7 wt/wt/wt to deliver 200 microg BDP, 6 microg FF, and 12.5 GB; 73.0: 8.8: 18.2 wt/wt/wt to deliver 50 microg BDP, 6 microg FF, and 12.5 GB; 80.3: 9.6: 10.1 wt/wt/wt to deliver 100 microg BDP, 12 microg FF, and 12.5 microg GB; 89.1: 5.3: 5.6 wt/wt/wt to deliver 200 microg BDP, 12 microg FF, and 12.5 microg GB; 72.7: 4.4: 22.9 to release 200 microg of BDP, 12 microg of FF and 63 microg of GB; 59.2: 3.5: 37.3 wt/wt/wt to deliver 100 microg BDP, 6 microg FF, and 63 microg GB, 76.3: 4.6: 19.1 wt/wt / weight to deliver 100 microg of BDP, 6 microg of FF, and 25 microg of GB and 84.4: 5.1: 10.5 weight/ weight/ weight to deliver 200 microg of BDP, 12 microg of FF, and 25 microg of GB.

[0091] In a preferred embodiment, the three active ingredients could be present in a ratio of 84.4: 5.1: 10.5, or 91.5: 2.7: 5.7, or 73.0: 8.8: 18.2 wt/wt/wt, more preferably 84.4: 5.1: 10.5 wt/wt/wt.

[0092] In preparation, the microparticles of the invention are shown to be chemically stable.

[0093] When released, they present a good constancy of the ratio of active ingredients and give rise to a high fraction of extrafine particles, indicating that they could be suitable for the prevention and/or treatment of diseases affecting the distal tract of the respiratory tree.

[0094] Furthermore, the microparticles of the invention have a regular and uniform spherical shape exhibiting more homogeneous adhesion forces together with the whole powder which are, in turn, associated with improved DPI performances.

[0095] The formatting factor is used to characterize the shape of microparticles.

[0096] Thus, the microparticles of the invention are characterized by means of a formatting factor comprised between 0.8 and 1.15, preferably between 0.9 and 1.10, more preferably between 0.95 and 1 .05.

[0097] The formatting factor could be determined according to the following equation reported in Kumar S et al., Curr Appl. Phys. Influence of metal powder shape on drag coefficient in a spray jet, 2009, 9, 678-682

[0098] where:

[0099] The RN indicates the roundness of the particle and is calculated by applying the following formula:

[00100] where p and A are the mean area and perimeter values, respectively, of at least ten spherical particles as measured from the Scanning Electron Microscopy (SEM) images.

[00101] Alternatively, the average area and perimeter can be measured using an optical microscope.

[00102] In Kumar S et al, it is reported that the formatting factors (SF) of circle is 1.00. It is also reported that the deviation from unity leads to particle irregularity, but particles with an SF value greater than 0.8 can be considered to have a spherical shape.

[00103] Scanning electron microscopy (SEM) or optical microscopy can also be used to qualitatively observe the characteristics of the powder particles of the Invention, such as the shape of the particles and their surface morphology.

[00104] Since the microparticles of the invention must be administered to the lungs by means of inhalation, at least 90% of them must have a volume diameter equal to or less than 6 microns.

[00105] On the other hand, it is well known that most available therapeutic methodologies tend to be associated with poor therapeutic control of individuals with respiratory diseases affecting the small airways, such as the phenotype of small airway asthma.

[00106] Through the microparticles of the invention, all three active ingredients could simultaneously reach the distal tract of the respiratory tree whereby they could act synergistically and improve the results of the small airways and associated control.

[00107] In reality, said microparticles are characterized by means of a well-defined, narrow and selected particle size distribution in which at least 90% of all of them having a volume diameter of less than 4.5 microns, preferably equal to or less than 4.0 microns, and its volume average diameter is comprised between 1.0 and 3.0 microns, more advantageously between 1.2 and 2.5 microns, preferably between 1.5 and 2, 2 microns.

[00108] Advantageously, not more than 10% of said microparticles having a volume diameter of less than 0.2 microns, preferably equal to or less than 0.5 microns, more preferably equal to or less than than 0.6 microns.

[00109] It follows that the width of the particle size distribution of the particles of each active ingredient, expressed as a range, should advantageously be comprised between 1.0 and 4.0, more advantageously between 1.2 and 3.5, of preferably between 1.5 and 2.0. Consequently Chew et al., J Pharm Pharmaceut Sci 2002, 5, 162-168, the range corresponds to [d (v, 0.9) - d (v,0,1)] / d (v,0,5) .

[00110] The size of the active particles is determined by measuring the characteristic equivalent sphere diameter, known as volume diameter, by means of laser diffraction. In the reported examples, the volume diameter has been determined using a Malvern mechanism. Anyway, another equivalent mechanism can be employed by the person skilled in the art.

[00111] When determined by the Brunauer-Emmett-Teller (BET) nitrogen absorption method according to a procedure known to the person skilled in the art, the Specific Surface Area of the microparticles of the invention will be comprised between 1.5 and 3 .5 m2/g, preferably between 2.0 and 3.0 m2/g, preferably between 2.2 and 2.8 m2/g.

[00112] The adhesion and cohesion forces of the microparticles of the invention were also evaluated by means of atomic force microscopy (AFM) according to the experimental procedure reported in Example 2. A layer of alpha lactose monohydrate powder was used to perform the measurements. This powder layer represents a heterogeneous surface in which the contact area for adhesion is highly variable. The contact area, in turn, is a dominant factor in determining the force of interaction. The comparison of the obtained cohesion and adhesion forces suggests that there is no statistical difference in the magnitude of the interaction of the microparticles of the invention with themselves or with the lactose powder. In sharp contrast, common micronized active ingredients are either powerfully adhesive or cohesive. By virtue of said properties, the microparticles of the invention can exhibit improved dispersion when formulated as interactive ordered mixtures with the excipient particles of lactose as a carrier. In fact, they would be less likely to give rise to the formation of the common micronized active ingredients like stable agglomerates.

[00113] In another aspect, the present invention provides a process for producing the microparticles of the invention comprising the steps of:

[00114] a) repairing a solution of the three active ingredients in a predetermined ratio in a suitable solvent;

[00115] b) generating an aerosol from the solution of said three active ingredients;

[00116] c) drying the atomized droplets to produce the microparticles; It is

[00117] d) isolating the produced microparticles.

[00118] As a result of said process, the microparticles are obtained as a completely amorphous powder in which each of the active ingredients is present in an amorphous form.

[00119] As far as step a) is concerned, the choice of solvent is critical as, in addition to having a high solubilizing capacity for the three active ingredients, it should have an adequate degree of diffusion and volatility characteristics within the atomized droplets. These properties actually significantly affect the particle size distribution of the resulting microparticles.

[00120] The person skilled in the art will be able to choose the solvent with respect to the desired particle size distribution.

[00121] Advantageously, the solvent could be selected from the group consisting of methanol, ethanol, water, DMSO, acetonitrile and mixtures thereof.

[00122] In a preferred embodiment of the invention, the solvent is a mixture of ethanol: water ranging from 85: 15 to 95: 5 volume/volume, preferably 90: 10 volume/volume.

[00123] For step b), any aerosol based on the atomization system could be employed to generate the aerosol. Various systems for generating aerosols are well known. The aerosol can, for example, be generated from the desired substance dissolved in a suitable solvent by means of electrohydrodynamic vaporization, high air pressure atomizer or other aerosol generators including pneumatic systems, spinning systems (spinning tops), air nozzles. sprayers, nebulizers, propellant evaporation systems, piezoelectric transducers and ultrasonic transducers.

[00124] In a preferred embodiment, the microparticles are prepared by spray drying. In this case the solution from step a) is introduced into the drying chamber of a vaporization dryer through an atomization device to form the droplets and the atomized droplets are dried by introducing a stream of preheated drying gas into the said drying chamber.

[00125] Any commercially available spray dryer can be advantageously employed.

[00126] The person skilled in the art will appropriately adjust the conditions of aerosol generation, such as the temperature of the solution, the flow rate of the solution and the pressure of the carrier gas in relation to the desired particle size distribution of the microparticles. them and the batch size.

[00127] In a specific embodiment of the invention, it could be preferred to obtain crystalline or partially amorphous microparticles in which at least one of the active ingredients is in crystalline form.

[00128] In this case, in step d) of the above-mentioned preparation process, the microparticles are collected in a container containing an antisolvent for all three active ingredients; and then a high intensity ultrasound is applied to change the morphology of the microparticles and cause the crystallization of at least one of the three active ingredients present in the microparticle. Finally the microparticles are isolated according to methods known to the person skilled in the art.

[00129] The antisolvent may advantageously be selected from the group consisting of n-heptane, cyclohexane, and fluorinated hydrocarbons such as perfluorodecalin.

[00130] Further details on the conditions to bring about crystallization are described in WO 2010/007447 and WO 2010/097188 the teachings of which are hereby incorporated by reference.

[00131] Crystalline or partially amorphous microparticles could also be isolated and collected.

[00132] When the microparticles of the invention comprising formoterol fumarate, beclomethasone dipropionate and glycopyrronium bromide are isolated as an amorphous powder, they contain all three active ingredients in anhydrous form.

[00133] On the contrary, when said microparticles are isolated as crystalline or partially amorphous powder, they could contain formoterol fumarate as dihydrate form and beclomethasone dipropionate as monohydrate form.

[00134] In a specific embodiment, all three active ingredients in the microparticles are in a crystalline form.

[00135] In this case, advantageously the extent of crystallinity, expressed as % by weight of the crystalline microparticle with respect to the total weight of the microparticle, is greater than 90%, preferably greater than 95%.

[00136] Amorphicity and/or crystallinity and its extent can be determined by employing X-ray powder diffraction or other techniques known to the skilled person, such as differential scanning calorimetry (DSC) or microcalorimetry.

[00137] The presence of all active ingredients in the microparticles could be detected by methods known to the skilled person, such as CP-MAR solid state C13 NMR spectroscopy Raman spectroscopy.

[00138] The microparticles of the invention are physically stable in storage for at least one month.

[00139] Anyway, in the case of partially amorphous or amorphous microparticles, further stabilization of the amorphous state can be obtained with the aid of suitable excipients in the relevant pharmaceutical formulation for inhalation, for example, mannitol, leucine, or trehalose .

[00140] Thus, in another aspect, the present invention provides a pharmaceutical formulation for administration via inhalation comprising the microparticles of the invention. Said microparticles may be formulated together with one or more pharmaceutically acceptable carriers, diluents, additives or excipients.

[00141] For example, the formulation can be provided in the form of a suspension in a propellant as an aerosol carrier to be administered by means of pressurized metered dose inhalers (pMDI).

[00142] The pMDI comprises a vessel in which the formulation is filled and a valve with a dosimeter for releasing a therapeutically effective dose per day of the formulation.

[00143] In certain embodiments, the aerosol carrier may consist of a non-chlorofluorocarbon based propellant, such as hydrofluoroalkane (HFA). In particular the propellants HFA 134a, and HFA 227 or mixtures thereof can be advantageously employed.

[00144] The suspension formulation may comprise additional excipients such as surfactants and wetting agents.

[00145] In a preferred embodiment, the formulation is provided in the form of a dry powder for inhalation, more preferably in the form of an ordered or interactive mixture, by diluting the particles of the invention in a pharmacologically inert physiologically acceptable excipient consisting of into coarser particles.

[00146] Advantageously, said powder formulation for inhalation may comprise the particles according to the Invention and coarse particles of a physiologically acceptable excipient, for example particles having an MMD greater than 90 microns and preferably the MD comprised between 50 microns and 500 microns, more preferably between 150 and 400 microns, even more preferably between 210 and 355 microns. In another embodiment, the coarse particles have an MD comprised between 90 and 150 microns.

[00147] In one of the preferred embodiments, when their MD is comprised between 210 and 355 microns, the coarse excipient particles preferably have a relatively highly fissured surface.

[00148] Preferably the above-mentioned powder formulation may also comprise a fraction of the particles having an MMD of less than 35 microns, preferably less than 15 microns, more preferably less than 10 microns, composed of the particles of a physiologically acceptable excipient. acceptable and particles of an additive material selected from the class of anti-adherents such as the amino acid isoleucine and leucine or from lubricants such as magnesium stearate, stearyl alcohol sodium stearyl fumarate, stearic acid and sodium monopalmitate sucrose (hereinafter the premix fraction).

[00149] The physiologically acceptable excipient may be constituted of any physiologically acceptable pharmacologically inert, amorphous or crystalline material of animal or vegetable source or combination thereof. Preferred materials are crystalline sugars and, for example, monosaccharides such as glucose or arabinose, or disaccharides such as maltose, trehalose, sucrose, dextrose or lactose. Polyalcohols such as mannitol, sorbitol, maltitol, lactitol can also be used. The most preferred material is α-lactose monohydrate.

[00150] Examples of commercial lactose are CapsulacTM, InhalacTM and PharmatoseTM. An example of commercial mannitol is PearlitolTM.

[00151] In a preferred embodiment, the fraction of the premix is composed of 98% by weight of α-lactose monohydrate and 2% by weight of magnesium stearate and the ratio between the fraction of microparticles and the fraction of the coarse particles made of α-lactose monohydrate particles is 10: 90% by weight, respectively.

[00152] The amount of magnesium stearate in the final formulation is advantageously comprised between 0.02% and 1.0% by weight on the total weight of the formulation, preferably between 0.05 and 0.5% by weight, more preferably between 0.1 and 0.4% by weight, even more preferably between 0.2 and 0.3% by weight.

[00153] The powder formulation for inhalation comprising the microparticles according to the Invention is endowed with good flow properties and is characterized by means of a high degree of homogeneity. In fact, after mixing, the uniformity of the active ingredient content, expressed as relative standard deviation (RSD), is less than 5%.

[00154] Said powder formulation can be administered via inhalation with any type of DPIs known in the art.

[00155] DPIs can be divided into two basic types: i) single-dose inhalers, for the administration of single pre-subdivided doses of the active compound; ii) multi-dose dry powder inhalers (MDPIs), either pre-subdivided single doses or pre-loaded with amounts of active ingredient sufficient for multiple doses. Based on the required inspiratory flow rates (l/min) which in turn are strictly dependent on their mechanical and planning characteristics, DPIs are divided into: i) low resistance devices (> 90 l/min); ii) medium resistance devices (about 60 l/min); iii) high resistance devices (about 30 l/min).

[00156] Dry powder formulations comprising the microparticles of the invention are particularly suitable for multi-dose DPIs comprising a reservoir from which individual therapeutic dosages can be withdrawn on demand by actuating the device, for example, the one described in WO 2004/012801.

[00157] Other multi-dose devices that can be used are, for example, the DISKUSTM by GlaxoSmithKline, the TURBOHALERTM by AstraZeneca, the TWISTHALERTM by Schering and the CLICKHALERTM by Innovata.

[00158] As marketed examples of single-dose devices, there may be mentioned ROTOHALERTM from GlaxoSmithKline, HANDIHALERTM from Boehringer Ingelheim, BreezehalerTM from Novartis, and Monodose RS01 from Plastiape.

[00159] In a preferred embodiment, the dry powder formulation is filled in the device with DPI described in WO 2004/012801, it is particularly suitable for the release of extrafine formulation.

[00160] To protect DPIs from moisture ingress into the formulation, it may be desirable to wrap the device in a flexible packaging capable of resisting moisture ingress, such as that described in EP 1760008.

[00161] The microparticles of the invention are indicated for the prevention and/or treatment of inflammatory or obstructive airway diseases, such as asthma and chronic obstructive pulmonary disease (COPD).

[00162] Other respiratory disorders characterized by obstruction of the peripheral airways as a result of inflammation and/or the presence of mucus, such as chronic obstructive bronchiolitis, bronchiectasis, and cystic fibrosis may also benefit from its use.

[00163] In certain embodiments, the microparticles of the invention are particularly suitable for the prevention and/or treatment of severe and/or very severe forms of respiratory disorders, in particular severe and/or very severe COPD and severe persistent asthma.

[00164] The invention is also illustrated by the following examples.

EXAMPLES Example 1 - Production of Microparticles according to the In-vention

[00165] 15 g of a mixture of formoterol fumarate dihydrate (FF), beclomethasone dipropionate (BDP) and glycopyrronium bromide (GB) in a ratio of 84.4: 10.5: 5.1% by weight were dissolved in 1500 ml of 90:10% v/v ethanol:water.

[00166] The feedstock solution was spray dried using a B290 benchtop spray dryer (Buchi) with a 2-fluid nozzle at a feed rate of 5 g min-1.

[00167] The spray drying parameters are reported in Table 1. TABLE 1

[00168] Upon completion of spray drying, the collection container and cyclone were separated from the B290 spray dryer and stored for 24 hours at a temperature of 25 °C and a relative humidity of 20%.

[00169] A flowable powder was recovered in a yield of 77% (11.7 g).

Example 2 - Characterization of the Microparticles of the Invention.

[00170] The microparticles as obtained in Example 1 were characterized in terms of drug content, particle size, morphology, physical state, adhesion/cohesion forces and specific surface area.

DRUG CONTENT

[00171] It was determined using the UPLC-PDA test (mean ± Standard Deviation; n = 5).

[00172] The results are reported in Table 2, indicating that they are within ± 3% of the target concentration for each active ingredient (API) in the spray-dried powder. TABLE 2

PARTICLE SIZE

[00173] The particle size was determined by means of laser diffraction using an R1 lens (Sympatec HELOS). A sample of the powder was dispersed for two measurement conditions employing an air pressure of 0.05 to 0.3 kPa (0.5 and 3 bar) respectively (Sympatec RODOS) and tested at a rate of 5 m/s from a humidity and temperature controlled dosing unit (Sympatec ASPIROS). Mean values d[v,10], d[v,50], d[v,90] were calculated from triplicate measurements.

[00174] The coverage was calculated using the following equation: Coverage = [d (v, 0.9) - d(v,0,1)]/ d(v,0,5)

[00175] The values obtained for the particle size, which are reported in Table 3, were not affected by the dispersion pressure, indicating a free-flowing powder without hard aggregates. TABLE 3

MORPHOLOGY

[00176] The morphology of the microparticles was determined by scanning electron microscopy (SEM) using the Jeol JSM-6480LV instrument.

[00177] The samples were mounted on carbon tape and stored in a vacuum for 12 hours before analysis to prevent the removal of adsorbed gases. Each sample was metallized with gold before imaging.

[00178] Formatting factor analysis was performed as follows. A sample of spray-dried powder was dispersed onto a 10 mm circular glass overslide and coated with gold using an agar-sprayed coater. SEM micrographs were obtained using a LEO1430VP instrument at an accelerating voltage of 10 kV and working distance of 10 mm. 120 SEM micrographs were acquired at 0° tilt and analyzed using Image Pro Analyzer Version 7.0. Measurement parameters were adjusted to acquire individual particles for shape analysis and any sampling anomalies, ie multiple particle selection, were manually removed. The rounding factor (RN) was determined using the equation RN = p2/4πA, where p is the perimeter and A is the area. The Particle Formatting Factor (PSF) was calculated from the rounding factor using the equation PSF = 1/RN. Statistical analysis was performed on the PSF and RN data to determine if enough particles were analyzed to give consistent data. The powder showed a uniform spherical morphology as demonstrated by the SEM images (Figure 1). Also the particle formatting and rounding factor (RN) determined by image analysis of the 3953 individual particles confirmed the spherical particle morphology (see Table 4). TABLE 4

PHYSICAL STATUS

[00179] It was investigated using X-ray powder diffraction (XRPD) and Differential Scanning Calorimetry (DSC).

[00180] The XRPD was performed using a Bruker AXS D8 Advance, equipped with a Vantec-1 detector and using Cu K-alpha radiation (1.54 A).

[00181] As for the DSC, the heat flux (W g-1) as a function of the temperature increase (°C) was determined using a TA Instruments Q2000 calorimeter. Samples were weighed into a non-airtight aluminum DSC crucible. Samples were equilibrated at 20°C before the temperature was increased by 5°C min-1 to 220°C. The analysis was performed in triplicate and interpreted using the Universal Analysis software.

[00182] DSC and XRPD also indicate that the material is amorphous. In fact, compared to the reference materials, the spray-dried microparticles did not show the peaks corresponding to the short range order and crystalline structure and typical amorphous halo is seen.

[00183] No melting point was identifiable in the DSC trace.

FORCES OF ADHESION / COHESION

[00184] The force measurement capabilities of the atomic force microscope (AFM) were employed to directly measure the adhesion forces between the two surfaces. Non-contact (Bruker) AFM cantilevers with constant calibrated spring (typically 0.2 - 0.4 Nm-1) were used for all adhesion - cohesion measurements. The probes were prepared by attaching the microparticles to the tip of the cantilever, which was confirmed by optical and variable pressure SEM prior to use. A layer of the spray-dried microparticles of Inhalac 50 lactose was fixed onto a silicon substrate using a thin layer of glue. The force-distance curves were recorded by monitoring the cantilever deflections as the probe and sample were brought into contact (methodological trace), and then separated (retract trace). Cohesion/adhesion data were obtained from microparticles and lactose samples consecutively with the same tip. Data were obtained on a 5 x 5 grid with 500 nm spacing. Each grid generates 25 force curves over a 2 x 2 μm area. The experiment was repeated with a different particle probe or different sampling area. All experiments were conducted under ambient conditions. A total of 153 cohesion and 144 adhesion strength curves were acquired and processed to extract the interaction force (nN).

[00185] The comparison of the obtained cohesion and adhesion forces suggests that there is no statistical difference in the magnitude of the interaction of the microparticles of the invention with themselves or with the lactose powder. In sharp contrast, common micronized active ingredients are either powerfully adhesive or cohesive. By virtue of said properties, the microparticles of the invention can exhibit improved dispersion when formulated as interactive ordered mixtures with the excipient particles of lactose as a carrier. In fact, they would be less likely to give rise to the formation of the common micronized active ingredients like stable agglomerates.

SPECIFIC SURFACE AREA

[00186] The specific surface area was determined by employing BET (Brunauer-Emmet-Teuer) of multiple nitrogen points with a TriStar II 3020 instrument.

[00187] The values are reported in Table 5. TABLE 5

Example 3 - Dry Powder Formulation Comprising the Microparticles of the Invention

[00188] A hard pellet carrier containing coarse lactose (sieve fraction 212 - 355 μm) and microchronized premix in a ratio of 9: 1 was prepared before production of the formulation in accordance with the teaching of WO 01/78693 . Briefly, the alpha-lactose monohydrate particles (212 - 355 µm sieve fraction) and a fraction of the alpha-lactose monohydrate and magnesium stearate premix were mixed in a 9:1 ratio.

[00189] A powder formulation (50 g batch size) containing 1.22% weight/weight of the microparticles of Example 1 equivalent to 100 μg of BDP, 12.5 μg of GB, and 6.0 μg of FF in a 10 mg dose, was prepared in a Turbula mixer. It was prepared by separately weighing the hard pellet carrier and the microparticles. Half of the vehicle was added to a stainless steel vessel followed by the microparticles. The remaining carrier was added and the contents mixed at 32 rpm for 90 minutes. The formulation was then sieved using a 500 µm sieve and mixed for an additional 30 minutes at 32 rpm. The formulation was stored in an amber glass vial for a minimum of 24 hours at 20°C and a relative humidity of 40% before further analysis.

Example 4

[00190] The powder formulation of Example 3 was characterized in terms of uniformity of distribution of active ingredients and aerosol performances after loading into the multi-dose dry powder inhaler described in WO 2004/012801, and cited hereinafter as NEXThaler ®.

DISTRIBUTION UNIFORMITY

[00191] About 10 mg of the formulation was weighed directly into a 25 mL volumetric flask. It has been tested 10 times. The flask was made up to volume with 60:40% v/v MeOH:H2O and sonicated for 2 minutes. Samples were analyzed for each active ingredient using the UPLC-PDA assay.

[00192] The results (mean value ± Standard Deviation) are reported in Table 6. TABLE 6

AEROSOL PERFORMANCES

[00193] The aerosol performances were determined using the Next Generation Impactor (NGI) with a USP induction port and pre-separator containing 15 mL of 60: 40% by volume/volume of MeOH: H2O. The critical flow (P3/P2 ratio) was < 0.5 at the sampling flow rate of 60 L/min. The aerodynamic particle size distribution was based on 5 actuations of the NEXThaler, each tested in 4 liters of air (equivalent to an inhalation time of 4 s). The device was weighed before and after each actuation to determine the injection weight (mg). A minimum of 1 minute was allowed between consecutive performances to allow the static charge to dissipate.

[00194] Samples were collected from the NGI employing a fixed volume technique. MeOH: H2O (60: 40% volume/vol) was dispensed into the induction port (including mouthpiece); pre-separator; and each step using an applicator with an electronic dosimeter. The induction port (30 mL) and pre-separator (50 mL) were sealed using silicone plugs and shaken by hand for 2 minutes before samples were collected. The NGI steps (10 ml of steps 1 - 2; 15 ml of step 3 - MOC) were shaken using an NGI shaker for 3 minutes. All samples were filtered using a 0.2 µm syringe filter prior to analysis via UPLC-PDA.

[00195] The following parameters were calculated: i) the released dose (DD) which is the amount of drug released from the device recovered in all parts of the impactor; ii) the fine particle mass (FPM) which is the amount of dose delivered having a particle size equal to or less than 5.0 microns iii) the fine particle fraction (FPF) which is the percentage of the particle dose slim; iv) the MMAD; and v) the extrafine FPP which is the percentage of the fine particle dose having a particle size equal to or less than 2.0 microns.

[00196] The results (mean value ± Standard Deviation, n = 6) are reported in Table 7. TABLE 7

[00197] From the data in Tables 6 and 7, it can be seen that the prepared formulation also shows excellent homogeneity with Standard Deviation values < 2% and high released dose and respirable fraction (FPF), for all three active ingredients .

[00198] They also give rise to a high fraction of particles having a diameter equal to or less than 2 microns (at least more than 35% for all active ingredients), indicating that it could be suitable for the prevention and/or the treatment of diseases affecting the distal tract of the respiratory tree.

Claims (12)

1. Multi-component microparticles, characterized in that they are for use in a formulation for inhalation, each microparticle comprising a combination of beclomethasone dipropionate, formoterol fumarate and glycopyrronium bromide, as active ingredients, in a ratio comprised between 35: 10: 55 and 94: 1: 5 weight / weight / weight, according to which said microparticles are defined by means of a formatting factor between 0.95 and 1.05 determined according to the following equation: where RN indicates the roundness of the particle and is calculated by applying the following formula: where p and A are the mean perimeter and area values, respectively, of at least ten spherical particles as measured from the Scanning Electron Microscopy (SEM) images; wherein at least 90% of all said microparticles have a volume diameter equal to or less than 4.5 microns, and the volume average diameter of said microparticles is comprised between 1.0 and 3.0 microns. 2. Microparticles, according to claim 1, characterized by the fact that no more than 10% of said microparticles have a volume diameter of less than 0.2 microns. 3. Microparticles, according to claim 1 or 2, characterized in that each microparticle consists of a combination of formoterol fumarate or its dihydrate form, glycopyrronium bromide and beclomethasone dipropionate. 4. Microparticles, according to any one of claims 1 to 3, characterized by the fact that each of the active ingredients is present in an amorphous form. 5. Process for the preparation of multicomponent microparticles as defined in claim 4, characterized in that it is for use in a formulation for inhalation, each microparticle comprising a combination of beclomethasone dipropionate, formoterol fumarate and glycopyrronium bromide, as active ingredients, in a ratio comprised between 35: 10: 55 and 94: 1: 5 weight/weight/weight, according to which said microparticles are defined by means of a formatting factor comprised between 0.95 and 1.05 determined according to the following equation: where RN indicates the roundness of the particle and is calculated by applying the following formula: where p and A are the mean perimeter and area values, respectively, of at least ten spherical particles as measured from the Scanning Electron Microscopy (SEM) images, and where the microparticles are obtained by a process comprising the steps of: a) preparing a solution of the three active ingredients in a predetermined ratio in a suitable solvent; b) generating an aerosol from the solution of said three active ingredients; c) drying the atomized droplets to produce the microparticles; and d) isolating the produced microparticles. 6. Process according to claim 5, characterized in that the solvent is selected from the group consisting of methanol, ethanol, water, DMSO, acetonitrile and mixtures thereof. 7. Pharmaceutical aerosol formulation for pressurized metered dose inhalers (pMDIs), characterized in that it comprises the microparticles, as defined in any one of claims 1 to 3, suspended in a liquefied propellant gas. 8. Pressurized metered dose inhaler (pMDI), characterized in that it comprises a canister filled with the pharmaceutical aerosol formulation, as defined in claim 7, and a dosimeter valve for releasing a therapeutically effective dose per day of the active ingredient. 9. Dry powder pharmaceutical formulation, characterized in that it comprises microparticles, as defined in any one of claims 1 to 3, and optionally a carrier. 10. Dry powder inhaler, characterized in that it is filled with the dry powder formulation, as defined in claim 9. 11. Microparticles, according to any one of claims 1 to 3, characterized in that they are for use in the prevention and/or treatment of an obstructive and/or inflammatory airway. 12. Microparticles, and according to claim 11, characterized by the fact that the disease is asthma or chronic obstructive pulmonary disease (COPD).