COMPOSITION COMPRISING AT LEAST ONE DRY POWDER OBTAINED BY SPRAY DRYING TO INCREASE THE STABILITY OF THE FORMULATION

Abstract

The present invention relates to inhalation formulations of drugs in the form of dry powder for inhalation administration deliverable as such with an inhaler and provided with high deliverability, respirability and stability. In particular, the invention relates to a pharmaceutical composition for inhalation use in powder form comprising a first powder comprising at least a powder (a1) comprising an active agent or a pharmaceutically acceptable salt thereof, in an amount greater than 1% by weight of the powder, leucine in an amount from 5 to 70% by weight of said powder, a sugar in an amount from 20 to 90% by weight of the powder;

and a second powder comprising a mixture of a first lactose which has an X50 from 35 to 75 μm, with a second lactose which has an X50 from 1.5 to 10 μm, the content of the first and second lactose in the mixture are respectively from 85% to 96% and from 4% to 15%. The ratio by weight between the first powder and the second powder is from 1/5 to 1/100, and the composition has a fine particle fraction (FPF) greater than 60% and a delivered fraction (DF) greater than 85%.

Claims

1. A pharmaceutical composition for inhalatory use in powder form, which comprises: a) a first powder comprising at least a powder (a.sub.1) comprising an active agent or a pharmaceutically acceptable salt thereof, in an amount greater than 1% by weight of said powder, leucine in amount from 5 to 70% by weight of said powder, a sugar in amount from 20 to 90% by weight of said powder; b) a second powder comprising a mixture of a first lactose which has an X50 from 35 to 75 μm, with a second lactose which has an X50 from 1.5 to 10 μm, the content of said first lactose and second lactose in said mixture being respectively from 85% to 96% and from 4% to 15%, wherein the ratio by weight of said first powder and said second powder is from 1/5 to 1/100, and said composition has a fine particle fraction (FPF) greater than 60% and a delivered fraction (DF) greater than 80%.

2. The composition according to claim 1, wherein said first powder comprises a third powder (a.sub.2) comprising leucine in an amount from 5 to 70% by weight of said third powder and lactose in an amount from 20 to 90% by weight of said third powder.

3. The composition according to claim 1, wherein said first powder comprises a fourth powder (a.sub.3) comprising an active agent or a pharmaceutically acceptable salt thereof, in an amount greater than 1% by weight of said fourth powder, leucine in an amount from 5 to 70% by weight of said fourth powder, and a sugar in an amount from 20 to 90% by weight of said fourth powder.

4. The composition according to claim 1, wherein said active agent is a hydrolyzable active agent.

5. The composition according to claim 1, wherein said active agent is selected from the group consisting of: short and long acting bronchodilator inhalers, corticosteroids, anti-cholinergics, antibiotics, mucolytics, heparin and its derivatives, substances with antioxidant action.

6. The composition according to claim 1, wherein said sugar is selected from the group consisting of: lactose, trehalose, sucrose and maltodextrin.

7. The composition according to claim 1, wherein said leucine is in an amount from 18 to 55% by weight.

8. The composition according to claim 1, wherein said sugar is in an amount from 40 to 80% by weight.

9. The composition according to claim 1, wherein said powders comprising leucine comprise a surfactant in an amount from 0.2 to 2% by weight of the powder.

10. The composition according to claim 7, wherein said surfactant is selected from the group consisting of benzalkonium chloride, cetrimide, docusate sodium, glyceryl monooleate, sorbitan esters, sodium lauryl sulfate, polysorbates, phospholipids, bile salts, block copolymers of polyoxyethylene and polyoxypropylene.

11. The composition according to claim 7, wherein said surfactant is in an amount from 0.4 to 0.8% by weight.

12. The composition according to claim 1, wherein said first powder has an X50 less than 5 μm.

13. The composition according to claim 1, wherein the content of said first lactose and second lactose in said mixture comprised in said second powder being respectively comprised from 91 to 95% and from 5 to 9%.

14. A kit for the administration of a drug as inhalation powder, comprising a metered amount of the composition according to claim 1 and a device for inhalation.

15. The composition according to claim 2, wherein said first powder comprises a fourth powder (a.sub.3) comprising an active agent or a pharmaceutically acceptable salt thereof, in an amount greater than 1% by weight of said fourth powder, leucine in an amount from 5 to 70% by weight of said fourth powder, and a sugar in an amount from 20 to 90% by weight of said fourth powder.

16. The composition according to claim 2, wherein said active agent is a hydrolyzable active agent.

17. The composition according to claim 3, wherein said active agent is a hydrolyzable active agent.

18. The composition according to claim 2, wherein said active agent is selected from the group consisting of: short and long acting bronchodilator inhalers, corticosteroids, anti-cholinergics, antibiotics, mucolytics, heparin and its derivatives, substances with antioxidant action.

19. The composition according to claim 3, wherein said active agent is selected from the group consisting of: short and long acting bronchodilator inhalers, corticosteroids, anti-cholinergics, antibiotics, mucolytics, heparin and its derivatives, substances with antioxidant action.

20. The composition according to claim 4, wherein said active agent is selected from the group consisting of: short and long acting bronchodilator inhalers, corticosteroids, anti-cholinergics, antibiotics, mucolytics, heparin and its derivatives, substances with antioxidant action.

Description

EXAMPLES

[0118] The methods for preparing the powders that constitute the pharmaceutical composition and for preparing the solid composition for use as diluent (hereinafter bulking agent) of the present invention will now be described.

[0119] Preparation of the Individual Powders.

[0120] The powders containing the active agents and the bulking agent were obtained by spray drying, a drying technique used to obtain powders with uniform and amorphous particles from solutions of active agents and excipients in appropriate solvent or mixture of solvents.

[0121] For the formulations described the solvents used are water and ethyl alcohol in a fixed ratio of 70/30. The concentration of dissolved solids is 1% w/v for formulations containing the active agent and 2% w/v for the bulking agent.

[0122] In the case of the powder containing as active agent Formoterol Fumarate, tiotropium bromide and bulking agent, all the components of the powder were dissolved in water and the solution thus obtained was added to the portion of ethyl alcohol slowly at 25° C., taking care not to cause precipitation of some of the components.

[0123] For the formulation containing Budesonide as active agent, the active agent was dissolved separately in the alcohol portion to which the aqueous solution of the excipients was added to obtain a single water-alcohol solution.

[0124] The water-alcohol solution thus obtained was processed by means of a Buchi Mod. B290 spray dryer, using an open cycle with the following parameters: [0125] nozzle diameter 0.7 mm [0126] atomization gas nitrogen [0127] atomization pressure 4 bar [0128] drying gas air [0129] aspiration 100% (35 m3/h) [0130] inlet temperature 170° C. [0131] feed speed 8% (2.4 ml/min)

[0132] Powder collection system: cyclone separator with glass collection vessel

[0133] Outlet filter: polyester sleeve.

[0134] At the end of the drying process the powder collection step was performed in controlled temperature and humidity conditions: temperature <25° C., relative humidity <35%.

[0135] The powders were packaged immediately after production in borosilicate glass vials inserted in a double aluminum foil bag heat-sealed under partial vacuum (30%).

[0136] Storage Conditions for Accelerated Stability Study.

[0137] The powders produced by spray drying, divided and packaged in borosilicate glass vials sealed inside in a double aluminum foil bag heat-sealed under partial vacuum (30%) were stored for an accelerated stability study in an oven at a temperature of 40° C. and relative humidity of 13%.

[0138] At each time interval established by the study, the samples corresponding to the stability point were taken, left to cool until reaching room temperature, opened in controlled conditions in a glove box (temperature<20° C., RH<35%) and analyzed as established in the protocol.

[0139] Characterization of the Powder: Particle Size Analysis.

[0140] The powders obtained after spray drying were characterized in terms of dry particle size using a Sympatec Helos light scattering device that analyzes the particle size according to the Fraunhofer theory and equipped with RODOS disperser.

[0141] The instrument was suitably calibrated with reference material and prepared following the instructions provided in the instrument user manual.

[0142] After appropriate cleaning before analysis, an amount of powder for each batch produced was analyzed without any preliminary preparation of the sample.

[0143] The dispersion gas used was compressed air suitably cleansed of particles.

[0144] The test method specified therefore provides for compliance with the following measures in relation to the sample, to the powder disperser and to the light scattering analyzer.

[0145] Sample [0146] size: about 100 mg [0147] feed procedure: with a spatula [0148] pre-treatment of the sample: none

[0149] RODOS Disperser [0150] Model M ID-NR 230 V/Hz 24Va [0151] Dispersion pressure: 3 bar

[0152] Light Scattering Analyzer [0153] Model: Helos [0154] Test method: Fraunhofer [0155] Software version: Windox 4.0 [0156] Test lens: R1 (0.1-35 μm) [0157] Minimum optical concentration: 1% [0158] Activation threshold: minimum optical concentration detectable 1% for max 30 seconds of time and with at least 100 ms of exposure of the sample.

[0159] All the tests were conducted in controlled temperature and humidity environments, temperature <25° C. and relative humidity <50% RH.

[0160] Size analysis provides volume median diameter (VMD) values of the population of particles in the sample of powder.

[0161] Characterization of the Powder: Residual Moisture Content.

[0162] The residual moisture content in the powder obtained by spray drying was measured using the Karl Fischer coulometric system method.

[0163] The C20 Compact Karl Fischer Coulometer Mettler Toledo titrator was used for this purpose, which uses as reagent HYDRANAL®-Coulomat AG.

[0164] The sample powders were accurately weighed in an amount of around 15-20 mg and the weight was recorded in the parameters of the sample. Titration was started immediately after adding the sample to the reagent bath.

[0165] At the end of the test, the instrument indicates directly the percentage of water contained in the sample.

[0166] Characterization of the Powder: Determination of Titer and Related.

[0167] Three different HPLC (High Performance Liquid Chromatography) methods were used to determine the content of active agents in samples from MSLI tests and in the formulations, as well as of their related substances, as set down below: [0168] Method 1: Determination of formoterol and budesonide in MSLI samples [0169] Determination of the titer of formoterol and budesonide [0170] Determination of the degradation products of formoterol and budesonide [0171] Method 2: Determination of tiotropium in MSLI samples (also in the presence of formoterol and/or budesonide) [0172] Method 3: Determination of the tiotropium titer (also in the presence of formoterol and/or budesonide) [0173] Determination of the degradation products (also in the presence of formoterol and/or budesonide)

[0174] Method 1

[0175] The test method used to determine the content in MSLI samples, titer and degradation products for formulations containing Formoterol/Budesonide, is characterized by the following parameters:

[0176] Solvent: 50/50 methanol/phosphate buffer pH 2.7 25 mM

[0177] Mobile phase: acetonitrile/phosphate buffer pH 2.9 2.82 mM [0178] gradient elution

TABLE-US-00001 Time % buffer Flow (min) % ACN pH 2.9 (ml/min) 0 22 78 0.5 2.5 22 78 0.5 3.0 41 59 0.7 8.0 41 59 0.7 10.0 70 30 0.7 12.0 22 78 0.6 15.0 22 78 0.6

[0179] Injection volume: 20 μL

[0180] Analysis column: Agilent Poroshell 120 EC-C18, 100 mm×3.0 mm, 2.7 μm

[0181] Column temperature: 30° C.

[0182] Wavelength: 220 nm (Formoterol Fumarate) and 240 nm (Budesonide)

[0183] Retention time: 2.4 min (Formoterol Fumarate) and 8.0 min (Budesonide)

[0184] An HPLC Agilent model 1200 with diode array type detector, model G1315C was used for the test.

[0185] The samples for analysis were obtained by dissolving in the solvent an amount of powder such as to obtain a concentration of 160 μg/ml for the Budesonide and 4.5 μg/ml for the Formoterol Fumarate, as for the reference solution.

[0186] The reference solution was injected three consecutive times before the sample to determine the precision of the system expressed as relative standard deviation percentage (RSD %), which must be less than 2%.

[0187] The active agent content is obtained by calculating the ratio of the areas with respect to the reference solution at known concentration. The degradation of the product is calculated as ratio between the sum of the areas of all the analysis peaks corresponding to the degradation products and the active agent taken as reference. All the analysis peaks whose chromatogram area was greater than 0.1% of the area of the active agent are counted in the sum of the degradation products.

[0188] Method 2

[0189] The test method used to determine the Tiotropium content in MSLI samples, alone or in combination with Formoterol and/or Budesonide, is characterized by the following parameters:

[0190] Solvent: 40/60 methanol/phosphate buffer pH 2.7 25 mM

[0191] Mobile phase: acetonitrile/phosphate buffer pH 2.9 2.82 mM [0192] gradient elution

TABLE-US-00002 Time % buffer Flow (min) % ACN pH 2.9 (ml/min) 0 22 78 0.5 2.9 22 78 0.5 3.3 22 78 1.0 4.0 22 78 1.0 4.1 41 59 0.7 9.0 41 59 0.7 11.0 80 20 0.6 13.0 22 78 0.6 16.0 22 78 0.6

[0193] Injection volume: 20 μL

[0194] Analysis column: Agilent Poroshell 120 EC-C18, 100 mm×3.0 mm, 2.7 μm

[0195] Column temperature: 30° C.

[0196] Wavelength: 220 nm (Formoterol Fumarate) and 240 nm (Tiotropium-Budesonide)

[0197] Retention time: 2.3 min Formoterol Fumarate; 3.5 min Tiotropium; 9.0 min Budesonide.

[0198] An HPLC Agilent model 1200 with diode array type detector, model G1315C was used for the test.

[0199] The reference solution was injected three consecutive times before the sample to determine the precision of the system expressed as relative standard deviation percentage (RSD %), which must be less than 2%.

[0200] The active agent content is obtained by calculating the ratio of the areas with respect to the reference solution at known concentration.

[0201] Method 3

[0202] The test method used to determine the titer and degradation products for formulations containing Tiotropium is characterized by the following parameters:

[0203] Solvent: 40/60 methanol/phosphate buffer pH 2.7 25 mM

[0204] Mobile phase: acetonitrile/phosphate buffer pH 2.9 2.82 mM [0205] gradient elution

TABLE-US-00003 Time % buffer Flow (min) % ACN pH 2.9 (ml/min) 0 20 80 0.7 6 20 80 1.0 15 25.6 74.4 1.0 15.5 25.6 74.4 1.2 18 32 68 1.2 25 40 60 1.2 28 60 40 1.2 29 60 40 1.4 33 70 30 1.4 35 70 30 0.7 40 20 80 0.7 60 20 80 0.7

[0206] Injection volume: 20 μL

[0207] Analysis column: Agilent Poroshell 120 EC-C18, 150 mm×4.6 mm, 2.7 μm

[0208] Column temperature: 30° C.

[0209] Wavelength: 240 nm-Tiotropium; 315 imp.F Tiotropium.

[0210] Retention time: 9 min Tiotropium;

[0211] An HPLC Agilent model 1200 with diode array type detector, model G1315C was used for the test.

[0212] The samples for analysis were obtained by dissolving in the solvent an amount of powder such as to obtain a concentration of 6 μg/ml for Tiotropium Bromide, as for the reference solution.

[0213] The reference solution was injected three consecutive times before the sample to determine the precision of the system expressed as relative standard deviation percentage (RSD %), which must be less than 2%.

[0214] The content in active agents is obtained by calculating the ratio of the areas with respect to the reference solution at known concentration. The degradation of the product is calculated as ratio between the sum of the areas of all the analysis peaks corresponding to the degradation products and the active agent taken as reference. All the analysis peaks whose chromatogram area was greater than 0.1% of the area of the active agent are counted in the sum of the degradation products.

[0215] Characterization of the Powder: Differential Scanning Calorimetry.

[0216] Differential scanning calorimetry or DSC is a thermoanalytical technique used to determine chemical and physical phenomena with endothermic or exothermic effect in a sample, such as variations in phase, loss of water, chemical reactions.

[0217] In DSC the sample is heated with constant heating speed and the amount of heat required to raise its temperature is a function of its thermal capacity. Each endothermic or exothermic phenomenon causes a reversible or irreversible change in the thermal capacity of the material and can be detected as a variation of the baseline of the thermogram.

[0218] Formulations containing amorphous lactose show during heating a typical decrease in thermal capacity corresponding to the glass transition of the lactose from amorphous solid state to a metastable state that rapidly leads to its crystallization, characterized by an exothermic peak.

[0219] The temperature corresponding to these phenomena varies as a function of the composition of the sample and of the environmental conditions in which the sample is stored and prepared.

[0220] The samples were prepared in a controlled environment (temperature <20° C., relative humidity 35-30%). 40 uL aluminum standard crucibles for DSC were filled with a weighed amount of powder between 1 mg and 3 mg and sealed with specific lid.

[0221] Calorimetry testing of the samples in question was carried out by subjecting the samples to a heating ramp from 20 to 200° C. with a temperature increase of 10° C./min.

[0222] The test gives a thermogram in which the thermal events that accompany progressive heating of the sample are visible.

[0223] The glass transition (Tg) is identifiable with a decreasing step, at times followed by an increase in the baseline caused by relaxation enthalpy. During evaluation of the thermograms the onset temperature of the phenomenon (Tg onset) is calculated, regardless of the sample size. The glass transition temperature is a stability index of the powder as it is a prelude to crystallization, which takes place above 100° C. The exothermic crystallization peak can be integrated and the area subtended by the curve is an index of the amorphous fraction of the sample.

[0224] Preparation of the Mixtures.

[0225] The formulations used for the aerosol characterization tests with MSLI were produced by mixing powders containing the active agents and bulking agent with the mixture of lactoses. The powders were mixed using an Ultra Turrax T10 mixer for a mixing time of 5 minutes considered sufficient for the 3.5 g of powder of the batches produced. Uniformity of the content was controlled with titer analysis on 10 samples taken from different points of the bulk.

[0226] The powders were divided in sealed vials and stored inside a double aluminum foil bag heat sealed with partial vacuum (30%).

[0227] The operations of mixing and dividing in vials were carried out inside a glove box in controlled humidity and temperature conditions; max temperature 20° C. and environmental relative humidity <35%.

[0228] Characterization of the Powder: Respirability Test with MSLI.

[0229] The Multi Stage Liquid Impinger (MSLI) is a device that simulates in vitro pulmonary deposition of an inhalation formulation. A inhalation formulation, delivered by appropriate inhaler and conveyed into the device by aspiration, is deposited in the various stages connected in series of the impactor as a function of its aerodynamic features, such as particle size, density, shape. Each stage of the MSLI corresponds to an interval of aerodynamic particle sizes of the powder deposited therein and the aerodynamic size distribution of the powder is obtained using HPLC testing to determine the amount of active agent in each stage, making it possible to calculate the median aerodynamic diameter and the respirable fraction, considered according to the European Pharmacopoeia with aerodynamic diameter <5.0 μm.

[0230] For the respirability test, the powders of the formulations of the examples were divided into Size 3 HPMC capsules and loaded from RS01 powder inhaler—model 7 single-dose, code 239700001AB (Plastiape S.p.A.).

[0231] The device was assembled following the instructions for use and the indications of the European Pharmacopoeia.

[0232] For test purposes, it is necessary to deliver 10 powder capsules for each respirability test. The tests can be conducted at different pressure drops. In the case of the inhaler RS01, the pressure drop of 2 KPa corresponds to a flow of 60±2 l/min for 4 seconds and the pressure drop of 4 KPa records a flow of 96±2 l/min for 2.4 s. derived from a pressure drop of 2 Kpa in the system.

[0233] The following aerodynamic diameter cut-offs correspond to this flow value for each stage.

TABLE-US-00004 Aerodynamic range diam. Aerodynamic range diam. Stage 2 KPa (60 ± 2 l/min) 4 KPa (96 ± 2 l/min) 1 >13 μm >10.3 μm 2 13.0-6.8 μm 10.3-5.4 μm 3 6.8-3.1 μm 5.4-2.5 μm 4 3.1-1.7 μm 2.5-1.3 μm 5 <1.7 μm <1.3 μm (filter):

[0234] The respirable fraction (Fine Particle Fraction) comprises particles having a median aerodynamic diameter of less than 5 μm and is calculated using specific software (CITDAS Copley).

[0235] The aerodynamic parameters of an inhalation formulation subjected to MSLI analysis are expressed in terms of: [0236] Delivered Fraction (DF): i.e. the percentage of the dose of active agent delivered from the mouthpiece of the inhaler [0237] Fine Particle Fraction (FPF): respirable fraction (aerodynamic diameter <5.0 μm) of active agent expressed as percentage of the amount delivered.

[0238] Quantitative determination of the active agent in each stage was performed by HPLC using the test method for titer and related.

Example 1

[0239] Example 1 was conducted producing formulations containing Formoterol Fumarate or Tiotropium Bromide, which are two active agents sensitive to the presence of free water in the formulation.

[0240] In the case of formoterol, formulations containing different amounts of leucine and lactose or mannitol were produced.

[0241] The example highlights the protective effect of lactose against formoterol, this protective effect is explained considering that lactose is capable of producing a scavenger effect against the free water present in the formulation.

[0242] To demonstrate this, formulations of 3 types were produced: [0243] A powder containing exclusively formoterol and leucine [0244] 2 powders with different lactose contents together with formoterol and leucine [0245] 2 powders containing formoterol and leucine in which lactose was substituted by a different sugar: mannitol

[0246] The formulations with lactose tend to acquire moisture over time, with consequent decrease of Tg, but degradation over time is limited. This limited degradation is presumably due to a scavenger effect produced by the lactose against the water, which is thus trapped in a rigid structure and prevented from reacting with the other components. Differently, the formulation without lactose which was already crystalline undergoes chemical degradation.

[0247] Of the two formulations containing lactose, the one with 50% is better, as it is more stable over time.

[0248] Formulations containing tiotropium, leucine and lactose at different concentrations of tiotropium were also produced to assess the minimum concentration of active agent in the formulation so as to obtain a stable powder.

TABLE-US-00005 TABLE 1A Tween Formoterol Tiotropium Leucine Lactose Mannitol 80 Water content (%) Ex. (%) (%) (%) (%) (%) (%) T0 T28 days 1 5.00 95.00 0.9 0.9 2 5.00 70.00 25.00 1.4 1.8 3 5.00 45.00 50.00 2.1 2.7 4 5.00 70.00 25.00 0.9 0.9 5 5.00 45.00 50.00 1.0 0.9 6 0.06 50.00 49.44 0.50 1.2 2.0 7 3.00 50.00 46.50 0.50 3.0 2.5 8 6.00 50.00 43.50 0.50 2.4 1.3

TABLE-US-00006 TABLE 1B Degradation products Tg (° C.) P. size (VMD) (%) Ex. T0 T28 days T0 T28 days T0 T28 days 1 Not detected Not detected 2.6 2.7 0.6 0.9 2 62.7 56.9 2.0 1.9 0.4 0.4 3 66.3 57.5 1.6 1.6 0.3 0.3 4 Not detected Not detected 2.3 2.2 0.2 1.6 5 Not detected Not detected 1.6 1.6 0.1 1.4 6 72.7 62.2 2.7 2.8 0.7 1.5 7 58.5 60.5 1.7 1.7 0.1 0.5 8 Not detected Not detected 1.7 1.7 0.3 0.5

Example 2

[0249] The example was conducted producing formulations containing as active agent Budesonide, defined as HLSA Bud, formulated with lactose and leucine (Table 3), formulations containing as active agent Formoterol Fumarate, defined as HLSA FF, formulated with lactose and leucine (Table 2).

[0250] The lactose powders used were Respitose® SV003 (DFE Pharma, Goch, D) e Lacto-Sphere® MM3 (Microsphere SA, Ponte Cremenaga, Lugano CH).

[0251] Identification of the optimal coarse/fine lactose ratio was based on the production of formulations with increasing amounts of LactoSphere MM3 in formulations containing HLSA FF, HLSA Bud and Respitose SV003 due to the aerodynamic characterization of each single formulation. The parameters evaluated through the MSLI test were the Fine Particle Fraction (FPF %) and the Delivered Fraction (DF %) in conditions with pressure drop of 4 KPa using the inhaler RS01 (Plastiape, Osnago, Lecco, I).

[0252] The results obtained show that a ratio of 91:9 Respitose SV003 (coarse lactose) and MM3 (fine lactose) guarantees high values of Delivered Dose (%) and high Fine Particle Fraction (%) respirability, at the same time ensuring that the mixture remains homogeneous over time.

TABLE-US-00007 TABLE 2 Powder containing Formoterol Fumarate (HLSA FF 2.25%) Formoterol Fumarate 2.25% Leucine 20.0% Lactose 77.25%  Tween 80  0.5%

TABLE-US-00008 TABLE 3 Powder containing Budesonide (HLSA Bud 8%) Budesonide 8.0% Leucine 50.0% Lactose 41.5% Tween 80 0.5%

TABLE-US-00009 TABLE 4 Powder containing Lactose Ex. Respitose SV003 Lactosphere MM3 9 100.0% 0.0% 10 98.0% 2.0% 11 94.0% 6.0% 12 91.0% 9.0% 13 90.0% 10.0% 14 85.0% 15.0% 15 80.0% 20.0% 16 70.0% 30.0%

TABLE-US-00010 TABLE 5 Lactose mix from Ex. HLSA FF HLSABDS Table 4 17 0.5% 2.5% From example 9 97% 18 0.5% 2.5% From example 10 97% 19 0.5% 2.5% From example 11 97% 20 0.5% 2.5% From example 12 97% 21 0.5% 2.5% From example 13 97% 22 0.5% 2.5% From example 14 97% 23 0.5% 2.5% From example 15 97% 24 0.5% 2.5% From example 16 97%

TABLE-US-00011 TABLE 6 DF % DF % FPF % FPF % Ex. Formoterol Budesonide Formoterol Budesonide 17 81.4% 85.3% 59.8% 51.5% 18 81.3% 87.0% 59.7% 49.1% 19 86.3% 91.3% 69.2% 68.7% 20 88.3% 90.6% 67.6% 69.6% 21 89.2% 92.8% 65.6% 63.8% 22 95.5% 97.4% 63.0% 66.5% 23 80.8% 79.6% 53.9% 66.8% 24 80.6% 80.5% 48.2% 63.1%

Example 3

[0253] Example 3 was conducted producing formulations containing as active agent Budesonide (defined as HLSA Bud in the table), formulated with lactose and leucine, formulations containing as active agent Formoterol Fumarate (defined as HLSA FF in the table), formulated with lactose and leucine and formulations containing as active agent Tiotropium (defined as HLSA Tio in the table) formulated with lactose and leucine. These formulations were mixed with a lactose powder containing a mixture of Repitose SV003 and of LactoSphere MM3.

[0254] Some formulations containing Formoterol and tiotropium at low percentages were also mixed with a powder containing lactose and leucine, in which lactose is used as filler to form a Bulking Agent (defined as BA in the table) or powder containing leucine and lactose but without active agent.

[0255] The powders contained in the composition according to the invention are as follows:

TABLE-US-00012 TABLE 7 Powder containing Formoterol Fumarate (HLSA FF 2.25%) Formoterol Fumarate 2.25% Leucine 20.0% Lactose 77.25%  Tween 80  0.5%

TABLE-US-00013 TABLE 8 Powder containing Formoterol Fumarate (HLSA FF 4.5%) Formoterol Fumarate 4.5% Leucine 20.0% Lactose 75.0% Tween 80 0.5%

TABLE-US-00014 TABLE 9 Powder containing Budesonide (HLSA Bud 8%) Budesonide 8.0% Leucine 50.0% Lactose 41.5% Tween 80 0.5%

TABLE-US-00015 TABLE 10 Powder containing Tiotropium (HLSA Tio 1.5%) Tiotropium 1.5% Leucine 50.0% Lactose 48.0% Tween 80 0.5%

TABLE-US-00016 TABLE 11 Powder containing Tiotropium (HLSA Tio 3%) Tiotropium 3.0% Leucine 50.0% Lactose 46.5% Tween 80 0.5%

TABLE-US-00017 TABLE 12 Powder containing Tiotropium (HLSA Tio 6%) Tiotropium 6.0% Leucine 50.0% Lactose 43.5% Tween 80 0.5%

TABLE-US-00018 TABLE 13 Lactose mix Respitose ® SV003 91% LactoSphere ® MM3  9%

TABLE-US-00019 TABLE 14 Bulking Agent (BA) Leucine .sup. 50% Lactose 49.5% Tween 80  0.5%

[0256] The powders were mixed according to the methods described above, in order to obtain formulations containing Budesonide and Formoterol, Tiotropium and mixtures thereof, in a dose of powder of 15 mg.

[0257] The parameters evaluated through the MSLI test were the Fine Particle Fraction (FPF %) and the Delivered Fraction (DF %) in conditions with pressure drop of 2 KPa using the inhaler RS01 (Plastiape, Osnago, Lecco,

TABLE-US-00020 TABLE 15 HLSA HLSA HLSA HLSA HLSA HLSA FF FF Tio Tio Tio Bud Lactose DF FPF Ex 2.25% 4.5% 1.5% 3% 6% 8% BA Mix % % 17 0.80% — — — — — — 99.20% 87.1% 40.8% 18 0.80% — — — — — 10.00% 89.20% 93.3% 60.4% 19 0.80% — — — — 8.00% 91.20% 88.4% 70.8% Formoterol Formoterol 89.9% 73.1% Budesonide Budesonide 20 — 0.40% — — — — — 99.60% 86.4% 50.5% 21 — 0.40% — — — — 10.00% 89.60% 85.0% 82.5% 22 — — — — — 8.00% — 92.00% 89.0% 74.4% 23 — — 2.67% — — — — 97.33% 88.3% 77.2% 24 — — — 1.33% — — — 98.67% 85.6% 70.7% 25 — — — 1.33% — — 10.00% 88.67% 93.9% 79.6% 26 — — — 1.33% — 8.00% — 90.67% 89.2% 73.3% Tiotropium Tiotropium 93.0% 74.0% Budesonide Budesonide 27 — — — — 0.67% — — 99.33% 82.4% 48.2% 28 — — — — 0.67% — 10.00% 89.33% 92.8% 69.4% 29 — — — — 0.67% 8.00% — 91.33% 89.2% 73.3% Tiotropium Tiotropium 93.0% 74.0% Budesonide Budesonide 30 0.80% — — 1.33% — 8.00% — 89.87% 89.9% 76.2% Tiotropium Tiotropium 95.0% 76.3% Budesonide Budesonide 89.8% 74.0% Formoterol Formoterol

Example 4

[0258] The example was conducted analyzing some products currently on the market containing Formoterol, Budesonide, Tiotropium or combinations thereof (Table 16). Crystalline mixtures of budesonide and formoterol (i.e. not formulated according to the present invention by spray drying) with lactose mixtures with different particle sizes according to the present invention (Table 17A and 17B) were also analyzed.

[0259] The products available on the market used for comparison were: [0260] Symbicort® produced by Astrazeneca with Budesonide/Formoterol Fumarate ratio expressed in μg of 160/4.5. [0261] Miflonide®—Budesonide 400 mcg, Novartis Farma S.p.A. —21040 Origgio (VA), Italy [0262] Foradil®—Formoterol Fumarate 12 mcg, Novartis Farma S.p.A.—21040 Origgio (VA), Italy [0263] Spiriva®—Tiotropium Bromide, 18 mcg, Boehringer Ingelheim, Italia S.p.A., (MI), Italy

[0264] The aerodynamic performance of the commercial products was assessed with the MSLI test conducted with a pressure drop of 4 KPa.

[0265] The example was conducted in order to assess the aerosol performance of the composition according to the present invention, emphasizing how this composition (see Example 3) can be administered maintaining a high dose of drug delivered through the mouthpiece and a percentage of fine particles able to ensure that the amount of drug deposited in the site of action is capable of performing the correct pharmacological action.

[0266] The aerodynamic performance of a dose of 15 mg of each formulation containing crystalline active agents was assessed with the MSLI test conducted at the pressure drop of 2 KPa and 60 l/min.

TABLE-US-00021 TABLE 16 FPF % FPF % FPF % Formoterol Budesonide Tiotropium Symbicort ® 35.6% 38.4% — 160/4.5 Foradil ® 33.8% — — Miflonide ® — 26.1% — Spiriva ® — — 54.8%

TABLE-US-00022 TABLE 17A Powder of crystalline Budesonide, Formoterol and Tiotropium C1 C2 C3 C4 Budesonide  0.64%  0.64% — — Micronized Ph. Eur., Industriale Chimica, S.r.l, Saronno, VA, Italy Formoterol  0.018% —  0.018% — Fumarate Dihydrate, Ph Eur. 7th Ed., Lusochimica, S.p.A., Lomagna, LC, Italy Tiotropium — — —  0.04% Bromide, Euroasian Chemicals PVT LTD, Mumbai - 400001 India Lactose Mix 99.342% 99.36% 99.982% 99.94% (according to Table XX)

TABLE-US-00023 TABLE 17B FPF % FPF % FPF % Formoterol Budesonide Tiotropium C1 2.1% 42.8% — C2 — 45.8% — C3 3.7% — — C4 — — 1.7%