BR112015025160B1 - TIOTROPIUM DRY POWDER - Google Patents

Abstract

TIOTROPIUM DRY POWDER. The present invention relates to respirable dry powder comprising respirable dry particles comprising sodium chloride, leucine, and tiotropium bromide, wherein the sodium chloride is about 60% to about 90%, the leucine is about 10% to about 40%, tiotropium bromide is from about 0.01% to about 0.5%, and, optionally, one or more additional therapeutic agents, up to about 20%, wherein all percentages are percentages by weight on a dry basis and all respirable dry particle components total 100%. The invention also relates to respirable dry powders containing respirable dry particles comprising sodium chloride, leucine, and tiotropium bromide, wherein the sodium chloride is 67% to 84%, the leucine is 12% to 28%. , tiotropium bromide is about 0.01% to about 0.5%, and, optionally, one or more additional therapeutic agents, up to about 20%, wherein all percentages are percentages by weight on a dry and all respirable dry particle components total 100%. The invention also relates to respirable dry powders containing respirable dry particles comprising about 79.5% to about 80.5% (w/w) chloride (...).

Description

RELATED REQUESTS

[0001] This application claims benefit of US Patent Application 61/925,400, filed on January 9, 2014, US Patent Application 61/874,146, filed on September 5, 2013 and Patent Application No. 61/807,063 , deposited on April 1, 2013; All teachings of these applications are incorporated herein by reference.

FUNDAMENTALS

[0002] The chemical structure of tiotropium was first described in the US Patent. 5,610,163 and RE 39,820. Salts of tiotropium include salts containing cationic tiotropium with one of the following anions: bromide, fluoride, chloride, iodine, C1-C4-alkylsulfate, sulfate, hydrogensulfate, phosphate, hydrogenphosphate, dihydrogen phosphate, nitrate, maleate, acetate, trifluoroacetate, citrate, fumarate, tartrate, oxalate, succinate and benzoate, C1-C4-alkylsulfonate, which may be optionally mono-, di- or tri-substituted by fluorine in the alkyl group, or phenylsulfonate which may optionally be mono- or poly-substituted by C1- C4-alkyl on the phenyl ring. Tiotropium bromide is an anticholinergic that provides therapeutic benefits, for example in the treatment of COPD and asthma, and is the active ingredient in SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) (Boehringer Ingelheim, Germany). Tiotropium bromide is known to crystallize in various forms, such as crystalline anhydrous form (described, for example, in US Patents 6,608,055; 7,968,717 and 8,163,913 (Form 11)), crystalline monohydrate (described, for example, in US Patents 6,777,423 and 6,908,928) and crystalline solvates (described, for example, in US Patent 7,879,871). The various crystalline forms of tiotropium can be distinguished by a number of different assays, including X-ray Powder Diffraction (XRPD), Differential Scanning Calorimetry (DSC), crystal structure, and infrared (IR) spectrum analysis. Tiotropium can be synthesized using a variety of methods that are well known in the art (including, for example, methods described in US Patents 6,486,321; 7,491,824; 7,662,963 and 8,344,143).

SUMMARY OF THE INVENTION

[0003] The present invention relates to respirable dry powder comprising respirable dry particles comprising sodium chloride, leucine and tiotropium bromide, wherein sodium chloride is from about 60% to about 90%, leucine is from about 10% to about 40%, tiotropium bromide is from about 0.01% to about 0.5%, and optionally one or more additional therapeutic agents, up to about 20%; Preferably, sodium chloride is about 67% to about 84%, leucine is about 12% to about 28%, tiotropium bromide is about 0.01% to about 0.5%. % and, optionally, one or more additional therapeutic agents, up to about 20%; more preferably, the sodium chloride is about 75% to about 82% and the leucine is about 15% to about 25%; and more preferably, the sodium chloride is about 79.5% to about 80.5% and the leucine is about 19.5% to about 20.5%, wherein all percentages are weight percentages. on a dry basis and all respirable dry particle components total 100%. The invention also relates to respirable dry powder comprising respirable dry particles comprising sodium chloride, leucine and tiotropium bromide, wherein sodium chloride is about 65% to about 86%, leucine is about 10 % to about 35%, tiotropium bromide is about 0.01% to about 0.5%, and optionally one or more additional therapeutic agents, wherein the one or more additional therapeutic agents is about 1 % to about 10%, more preferably, one or more additional therapeutic agents is from about 3% to about 7%, and more preferably, one or more additional therapeutic agents is from about 4% to about 5%, wherein all percentages are weight percentages on a dry basis and all respirable dry particle components total 100%.

[0004] The invention also relates to respirable dry powder comprising respirable dry particles consisting of sodium chloride, leucine and tiotropium bromide, wherein the ratio of sodium chloride to leucine is 2.5:1 to 8:1 (w/w), tiotropium bromide is about 0.01% to about 0.5%, and, optionally, one or more additional therapeutic agents up to about 20%; preferably, the ratio of sodium chloride to leucine is 3:1 to 6:1 (w/w); and more preferably, the ratio of sodium chloride to leucine is about 4:1 (w/w), where all percentages are weight percentages on a dry basis and all respirable dry particle components total 100%. The invention also relates to respirable dry powder comprising respirable dry particles consisting of sodium chloride, leucine and tiotropium bromide, wherein the ratio of sodium chloride to leucine is 1.5:1 to 9:1 (w/ p), tiotropium bromide is from about 0.01% to about 0.5%, and, optionally, one or more additional therapeutic agents up to about 20%; Preferably, the ratio of sodium chloride to leucine is 1.9:1 to 8.5:1 (w/w), wherein all percentages are weight percentages on a dry basis and all components of dry respirable particles total 100%.

[0005] Respirable dry powder comprising dry respirable particles sometimes does not contain additional therapeutic agent.

[0006] Respirable dry powder comprising dry respirable particles, on other occasions, contains additional therapeutic agent of 20% or less of the formulation by dry weight. The one or more additional therapeutic agents may be present in an amount of about 0.01% to 10%, specifically, the one or more additional therapeutic agents may be present in an amount of about 0.01% to 0.5%. %, in an amount greater than 0.5% to 3%, or an amount greater than 3% up to about 10%. The one or more additional therapeutic agents are independently selected from the group consisting of one or more corticosteroids, one or more long-acting beta-agonists, one or more short-acting beta-agonists, one or more anti-inflammatory agents, one or more bronchodilator and any combination thereof.

[0007] Respirable dry powder comprising respirable dry particles contains from about 0.01% to about 0.5% tiotropium bromide or contains from about 0.02% to about 0.25% tiotropium bromide, where all percentages are weight percentages on a dry basis.

[0008] Respirable dry powder comprising dry respirable particles has one or more of the following characteristics and/or properties: a volume mean geometric diameter (VMGD) of about 10 micrometers or less, or from about 1 micrometer to about 4 micrometers ; a compacted density of at least about 0.45 g/cm3, of about 0.45 g/cm3 to about 1.2 g/cm3, of at least about 0.5 g/cm3, of at least about from 0.55 g/cm3, or from at least about 0.55 g/cm3 to about 1.0 g/cm3; a mass average aerodynamic diameter of between about 1 micron to about 5 microns, of between about 2 microns and about 5 microns, or preferably, of between about 2.5 microns and about 4.5 microns; a fine particle dose of less than 4.4 microns of between about 1 microgram and about 5 micrograms of tiotropium, or of about 2.0 micrograms and about 5.0 micrograms of tiotropium; a ratio of fine particle dose less than 2.0 microns to fine particle dose less than 4.4 microns less than about 0.50, or preferably, less than about 0.35, or less than about 0.25; a 1/4 bar dispersibility ratio of less than about 1.5, or less than about 1.4, a 0.5/4 bar dispersibility ratio of about 1.5 or less, all dispersibility were measured by laser diffraction; a total dose FPF of less than 3.4 microns of about 25% or more, of about 25% to about 60%, of about 40% or more, or of about 40% to about 60 %; a total dose FPF of less than 5.6 microns of about 45% or more, of about 45% to about 80%, of about 60% or more, or of about 60% to about 80%.

[0009] Furthermore, they may have a compacted density of at least about 0.4 g/cm3, a compacted density greater than 0.4 g/cm3, or greater than 0.4 g/cm3 at about 1.2 g/cm3; a mass average aerodynamic diameter (MMAD) preferably between about 3 microns and about 5 microns, or about 4 microns; or from about 3 microns to about 6 microns, or, preferably, from about 4 microns to about 6 microns, preferably, from about 5 microns; a fine particle dose of less than 4.4 microns of between about 1 microgram and about 4 micrograms of tiotropium, or preferably, between about 1.5 micrograms and 3.5 micrograms, about 2.0 micrograms, about 2.5 micrograms or about 3.0 micrograms; a ratio of fine particle dose less than 2.0 microns to fine particle dose less than 5.0 microns of less than about 0.50 or, preferably, less than about 0.35, or preferably less than 0.25, less than 0.20 or less than 0.18.

[0010] Respirable dry powder comprising respirable dry particles has a capsule emitted powder mass of at least 80% when emitted from a passive dry powder inhaler having a resistance of about 0.036 sqrt(kPa)/liters per minute under the following conditions; an inhalation energy of 2.3 joules at a flow rate of 30 LPM using a capsule of size 3, which contains a total mass of 10 mg, said total mass, which consists of dry respirable particles, and at which mean geometric diameter of volume of dry respirable particles emitted from the inhaler, as measured by laser diffraction, is 5 microns or less.

[0011] The invention also relates to a method for treating a respiratory disease and/or to a method for treating or reducing the incidence or severity of an acute exacerbation of a respiratory disease, by administering to the respiratory tract of a patient in need. of the same, respiratory dust comprising respiratory particles.

[0012] Furthermore, the invention also relates to a method for alleviating the symptoms of a respiratory disease and/or a method for improving the lung function of a patient with a respiratory disease, by administering to the respiratory tract of a patient in need. thereof, respiratory dust comprising respiratory particles. Without wishing to be bound by theory, it is believed that improving a patient's lung function over a period of time is a long-term way to treat respiratory disease and/or prevent acute exacerbations.

[0013] The respirable dry powder comprising respirable dry particles can be used 1) in the treatment of a respiratory disease in an individual, the use comprising administering to the respiratory tract of the individual an effective amount of the respirable dry powder, resulting in the treatment of a respiratory disease , and/or, 2) in treating or reducing the incidence or severity of an acute exacerbation of a respiratory disease in an individual, the use comprising administering to the respiratory tract of the individual an effective amount of the dry respirable powder, resulting in the treatment or reduction the incidence or severity of an acute exacerbation of a respiratory disease.

[0014] Furthermore, the respirable dry powder comprising respirable dry particles can be used 1) in relieving the symptoms of a respiratory disease, the use comprising administering to the respiratory tract of the individual an effective amount of the respirable dry powder, resulting in the relief of the symptoms of a respiratory disease, and/or, 2) in improving the lung function of a patient with a respiratory disease, the use comprising administering to the individual's respiratory tract an effective amount of the dry respirable powder, resulting in an improvement of the lung function of a patient with a respiratory illness.

[0015] In some embodiments, the respiratory disease is COPD, chronic bronchitis, emphysema, asthma, cystic fibrosis or non-cystic fibrosis bronchiectasis. The respiratory disease is preferably COPD, chronic bronchitis, and/or emphysema.

[0016] Respirable dry powder comprising respirable dry particles can be contained in a dry powder inhaler. The dry powder inhaler may be a capsule-based dry powder inhaler, a blister-based dry powder inhaler or a reservoir-based dry powder inhaler. The respirable dry powder comprising respirable dry particles may be contained in a container. The container may be a capsule or a blister, wherein the container is suitable for any of the dry powder inhalers listed above. The container contains dry respirable powder of a mass of about 15 mg or less, about 11 mg or less, about 8.5 mg or less, about 6 mg or less, or about 4 mg or less. The container may contain dry respirable powder of a mass of about 15 mg, 10 mg, 7.5 mg, 5 mg, 2.5 mg or 1 mg. The container may contain a nominal dose of tiotropium from about 1.5 to about 12 micrograms, from about 3 to about 12 micrograms, from about 3 to about 9 micrograms, or from about 3 to about 6 micrograms. micrograms. The container may contain a nominal dose of tiotropium of about 1.5 micrograms, about 3 micrograms, about 6 micrograms, about 9 micrograms, or about 12 micrograms. The container may be contained in a dry powder inhaler or may be packaged and/or sold separately.

[0017] Respirable dry powder comprising respirable dry particles can be contained in a dry powder inhaler. The dry powder inhaler is preferably a capsule-based dry powder inhaler. More preferably, the dry powder inhaler is selected from the RS01 family of dry powder inhalers (Plastiape S.p.A., Italy). More preferably, the dry powder inhaler is selected from the RS01 HR or RS01 UHR2. In one respect, the dry powder inhaler is not the RS01 HR. Most preferably, the dry powder inhaler is the RS01 UHR2. The respirable dry powder comprising respirable dry particles may be contained in a container. The container is preferably a capsule. Preferably, the capsule material is selected from gelatin and HPMC (hydroxypropylmethylcellulose). More preferably, the capsule material is HPMC. In one aspect, the capsule material is not gelatin. Preferably, the container is a size 3 capsule. More preferably, the container is a size 3 HPMC capsule. More preferably, respirable dry powder comprising respirable dry particles is contained in a size 3 HPMC capsule for use in the inhaler. of dry powder RS01 UHR2. In one aspect, respirable dry powder consists of respirable dry particles.

BRIEF DESCRIPTION OF FIGURES

[0018] Figure 1 A) is a graph representing the geometric mean of plasma tiotropium levels in pg/mL over time for exemplary Formulations I, II, III, and IV, respectively. B) is a graph illustrating the area under the curve (AUC) for hours 0-2 after administration of Formulations I, II, III, and IV, respectively, as (pgtiotropium per hour)/mL serum.

[0019] Figure 2 is a graph representing the average change in FEV1 (forced expiratory volume in the first second) from baseline over time for exemplary Formulations I, II, III, IV and placebo, respectively.

[0020] Figure 3 is a graph of the aerodynamic size distributions at a pressure drop of 4 kPa for Formulation II at a nominal dose of 5.8 μg and SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) at a dose nominal 18 μg, illustrating a similar fine particle dose (FPD) to Formulation II with a reduced nominal drug load.

[0021] Figure 4 is a graph showing that Formulation II has minimal oral deposition versus SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), reducing potential side effects.

[0022] Figure 5 is a graph illustrating the dependence of the reduced flow rate of Formulation II using the RS01 versus SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) over a range of patient-relevant inhalation pressure drops.

[0023] Figure 6 is a graph representing the geometric mean of plasma tiotropium levels in pg/mL over time for exemplary Formulations I, II, III and SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), respectively .

[0024] Figure 7 is a graph depicting the average change in FEV1 (forced expiratory volume in the first second) from baseline over time for exemplary Formulations I (nominal dose of 3 micrograms, also called “3μg”) , II (nominal dose of 6 micrograms, also called “6μg”), III (nominal dose of 9 micrograms, also called “9μg”), SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) and placebo.

[0025] Figure 8 is a graph of aerodynamic size distributions at a pressure drop of 4 kPa for Formulation II released from RS01 UHR2 at a nominal dose of 5.8 μg and SPIRIVA (tiotropium bromide) HANDIHALER (inhaler dry powder) at a nominal dose of 18 μg, illustrating a similar fine particle dose (FPD) to Formulation II with a reduced nominal drug load.

[0026] Figure 9 is a graph showing that Formulation II released from RS01 UHR2 has minimal oral deposition versus SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), reducing potential side effects.

[0027] Figure 10 is a graph illustrating the dependence of the reduced flow rate of Formulation II released from the RS01 UHR2 versus the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) over a range of relevant inhalation pressure drops of the patient.

DETAILED DESCRIPTION OF THE INVENTION

[0028] The present invention relates to respirable dry powders containing respirable dry particles comprising sodium chloride, leucine and tiotropium bromide, wherein sodium chloride is from about 60% to about 90%, leucine is from about 10% to about 40%, tiotropium bromide is from about 0.01% to about 0.5%, and, optionally, one or more additional therapeutic agents up to about 20%; Preferably, sodium chloride is about 67% to about 84%, leucine is about 12% to about 28%, tiotropium bromide is about 0.01% to about 0.5%. % and, optionally, one or more additional therapeutic agents up to about 20%; preferably, the sodium chloride is about 75% to about 82% and the leucine is about 15% to about 25%; and more preferably, the sodium chloride is about 79.5% to about 80.5% and the leucine is about 19.5% to about 20.5%, wherein all percentages are percentages in weight on a dry basis and all respirable dry particle components total 100%. The invention also relates to respirable dry powder comprising respirable dry particles comprising sodium chloride, leucine and tiotropium bromide, wherein sodium chloride is about 65% to about 86%, leucine is about 10 % to about 35%, tiotropium bromide is about 0.01% to about 0.5%, and optionally one or more additional therapeutic agents, wherein the one or more additional therapeutic agents is about 1 % to about 10%, more preferably, one or more additional therapeutic agents is from about 3% to about 7%, and more preferably, one or more additional therapeutic agents is from about 4% to about 5%, wherein all percentages are weight percentages on a dry basis and all respirable dry particle components total 100%.

[0029] The invention also relates to respirable dry powder comprising respirable dry particles consisting of sodium chloride, leucine and tiotropium bromide, wherein the ratio of sodium chloride to leucine is 2.5:1 to 8:1 (w/w), tiotropium bromide is from about 0.01% to about 0.5%, and, optionally, the one or more additional therapeutic agents up to about 20%; Preferably, the ratio of sodium chloride to leucine is 3:1 to 6:1 (w/w); and more preferably, the ratio of sodium chloride to leucine is about 4:1 (w/w), where all percentages are weight percentages on a dry basis and all respirable dry particle components total 100%. The invention also relates to respirable dry powder comprising respirable dry particles consisting of sodium chloride, leucine and tiotropium bromide, wherein the ratio of sodium chloride to leucine is 1.5:1 to 9:1 (w/ p), the tiotropium bromide is from about 0.01% to about 0.5%, and, optionally, the one or more additional therapeutic agents up to about 20%; Preferably, the ratio of sodium chloride to leucine is 1.9:1 to 8.5:1 (w/w), wherein all percentages are weight percentages on a dry basis and all components of dry respirable particles total 100%.

[0030] These respirable dry powders comprising respirable dry particles can be manufactured from their components, in solutions or suspensions that are aqueous and/or contain another solvent, by spray drying or other comparable processes. Respirable dry powders comprising respirable dry particles are relatively dry in water and solvent content, small in geometric diameter, dense in mass density, and dispersible so that they deagglomerate from each other with a relatively low amount of energy. They have superior aerosol properties, such as a relatively small aerodynamic diameter, a relatively high fine particle fraction, and fine particle dose below the sizes that are relevant to pulmonary deposition. These properties are illustrated by two exemplary Formulations in Example 1. Three additional preferred respirable dry powders comprising respirable dry particles comprising sodium chloride, leucine and tiotropium are described in Example 3. Despite the processing that occurs to prepare the respirable dry powders comprising particles In respirable dry conditions, tiotropium bromide maintains its activity as demonstrated in the in vivo experiment described in Examples 2 and 4.

Definitions

[0031] As used herein, the term “about” refers to a relative range of plus or minus 5% of a given value, for example, “about 20 mg” would be “20 mg plus or minus 1 mg” .

[0032] As used herein, the terms “administration” or “administering” dry respirable particles refers to the introduction of dry respirable particles into the respiratory tract of a subject.

[0033] The term “capsule emitted powder mass” or “CEPM” as used herein refers to the amount of dry powder formulation emitted from a capsule or unit dose container during an inhalation maneuver. CEPM is measured gravimetrically, typically by weighing a capsule before and after an inhalation maneuver to determine the mass of powder formulation removed. CEPM can be expressed as the mass of dust removed, in milligrams, or as the percentage of initial dust mass introduced into the capsule before the inhalation maneuver.

[0034] The term “dispersible” is a term of art that describes the characteristics of a dry powder or dry particles to be dissipated in a respirable aerosol. Dispersability of a powder or dry particles is here expressed as the quotient of the volume mean geometric diameter (VMGD) measured at a dispersing pressure (i.e. regulator) of 1 bar, divided by the VMGD measured at a dispersing pressure (i.e. , regulator) of 4 bar, or VMGD at 0.5 bar divided by the VMGD at 4 bar, as measured by laser diffraction, such as with HELOS/RODOS. These ratios are referred to herein as “dispersibility ratio 1 bar/4 bar”, and “dispersibility ratio 0.5 bar/4 bar”, respectively, and dispersibility correlates with a low ratio. For example, the 1 bar/4 bar dispersibility ratio refers to the VMGD of dry respirable particles or powders emitted from the orifice of a RODOS dry powder disperser (or equivalent technique) at about 1 bar, as measured by HELOS or another laser diffraction system, divided by the VMGD of the same particles or respirable dry powders measured at 4 bar by HELOS/RODOS. Thus, a highly dispersible dry powder or respirable dry particles will have a 1 bar/4 bar dispersibility ratio or 0.5 bar/4 bar dispersibility ratio that is close to 1.0. Highly dispersible powders have a low tendency to clump, clump or clump together and/or, if clumped, clumped or clumped together, are easily dispersed or clumped together as they are emitted from an inhaler and are breathed in by a subject. Dispersibility can also be assessed by measuring the size emitted from an inhaler as a function of flow rate. As the flow rate through the inhaler decreases, the amount of energy available in the air stream to be transferred to the powder to disperse it decreases. A highly dispersible powder will have its size distribution as aerodynamically characterized by its mass mean aerodynamic diameter (MMAD) or geometrically by its VMGD, not increase substantially over a flow rate range typical of human inhalation, such as about 15 to 60 LPM, or about 20 to 60 LPM. VMGD may also be called volume mean diameter (VMD), x50, or Dv50.

[0035] The term “dry particles” as used herein refers to respirable particles that may contain up to about 15% water or other solvent. Preferably, the dry powders contain water or other solvent up to about 10%, up to about 5%, up to about 1%, or between 0.01% and 1%, by weight of dry particles, or may be substantially free of water or other solvent or be anhydrous.

[0036] The term “dry powders” as used herein refers to compositions comprising dry respirable particles. Preferably, the respirable dry powders contain water or other solvent up to about 10%, up to about 5%, up to about 1%, or between 0.01% and 1%, by weight of dry respirable particles, or may be substantially free of water or other solvents, or be anhydrous.

[0037] The term “effective amount”, as used herein, refers to the amount of agent necessary to achieve the desired effect; improve or prevent deterioration of respiratory function (e.g., improve forced expiratory volume in one second (FEV1) and/or forced expiratory volume in one second (FEV1) as a ratio of forced vital capacity (FEV1/FVC)), reduce the occurrence of an acute exacerbation of a respiratory disease, e.g. Chronic Obstructive Pulmonary Disease (COPD), asthma, cystic fibrosis (CF) and non-cystic fibrosis bronchiectasis.

[0038] The actual effective amount for a particular use may vary according to the particular dry powder or dry particle, the mode of administration, and the age, weight, general health of the subject and the severity of the symptoms or condition to be be treated. The appropriate amounts of dry powders and dry particles to be administered and dosage schedules for a particular patient can be determined by a skilled clinician based on these and other considerations.

[0039] As used herein, the term “emitted dose” or “ED” refers to an indication of release of a drug formulation from a suitable inhaler device following a trigger or dispersion event. More specifically, for dry powder formulations, the ED is a measure of the percentage of powder that is removed from a unit dose container and exits the mouthpiece of an inhaler device. ED is defined as the ratio of dose delivered by an inhaler device to nominal dose (i.e., the mass of powder per unit dose placed in a suitable inhaler device before firing). ED is an experimentally measured parameter and can be determined using the USP Method Section 601 Aerosols, Metered Dose Inhalers, and Dry Powder Inhalers, Uniformity of Delivered Dose, Sampling of Delivered Dose of Dry Powder Inhalers, Convention of the United States Pharmacopoeia Unidos, Rockville, MD, 13th Revision, 222-225, 2007. This method uses an in vitro device designed to mimic patient dosing.

[0040] The terms “FPF (<5.6)”, “FPF (<5.6 microns)”, and “fine particle fraction of less than 5.6 microns”, as used herein, refer to the fraction of a sample of dry particles that have an aerodynamic diameter of less than 5.6 microns. For example, FPF (<5.6) can be determined by dividing the mass of dry respirable particles deposited in phase two and on the final collection filter of a two-phase collapsed Andersen Cascade Impactor (ACI) by the mass of heavy respirable dry particles within a capsule for release into the instrument. This parameter can also be identified as “FPF_TD (<5.6)”, where TD means total dose. A similar measurement can be conducted using an eight-phase ACI. The eight-phase ACI cutoffs are different from the 60 L/min flow rate standard, but the FPF_TD (<5.6) can be extrapolated from the full eight-phase data set. The result of the eight-phase ACI can also be calculated using the USP method of using the dose collected in the ACI instead of that in the capsule to determine FPF.

[0041] The terms “FPD (<4.4)”, “FPD <4.4 μm”, “FPD (<4.4 microns)” and “fine particle dose of less than 4.4 microns”, such as used herein, refer to the mass of respirable dry dust particles that have an aerodynamic diameter of less than 4.4 micrometers. For example, FPD <4.4 μm can be determined using an eight-stage ACI at a standard flow rate of 60 L/min and summing the mass deposited on the final collection filter, and stages 6, 5, 4, 3 and 2 for a single dose of powder activated in the ACI.

[0042] The terms “FPF (<5.0)”, “FPF <5 μm”, “FPF (<5.0 microns)”, and “fraction of fine particles of less than 5.0 microns”, such as used herein, refer to the fraction of a mass of dry respirable particles that have an aerodynamic diameter of less than 5.0 micrometers. For example, FPF (<5.0) can be determined using an eight-phase ACI at standard flow rate of 60 L/min from extrapolation of the full eight-phase data set. This parameter can also be identified as “FPF_TD (<5.0)”, where TD means total dose. When used in conjunction with a geometric size distribution, such as those indicated by Malvern Spraytec, Malvern Mastersizer or Sympatec HELOS Particle Sizer, “FPF (<5.0)” refers to the mass fraction of dry respirable particles that have a diameter geometric size of less than 5.0 micrometers.

[0043] The terms “FPF (<3.4)”, “FPF (<3.4 microns)”, and “fine particle fraction of less than 3.4 microns”, as used herein, refer to the fraction of a mass of dry respirable particles that have an aerodynamic diameter of less than 3.4 microns. For example, the FPF (<3.4) can be determined by dividing the mass of dry respirable particles deposited on the final collection filter of the two-phase collapsed ACI by the total mass of dry respirable particles weighed within a capsule for release on the instrument. This parameter can also be identified as “FPF_TD (<3.4)”, where TD means total dose. A similar measurement can be conducted using an eight-phase ACI. The eight-phase ACI result can also be calculated using the USP method of using the dose collected in the ACI instead of that in the capsule to determine the FPF.

[0044] “Hausner Ratio” is a technical term that refers to the compacted density divided by the bulk density and typically correlates with the fluidity of the bulk powder (i.e., an increase in the Hausner Ratio typically corresponds to a decrease in powder fluidity).

[0045] The term “respirable”, as used herein, refers to dry particles or dry powders that are suitable for release into the respiratory tract (e.g., pulmonary release) of a subject by inhalation. Dry powders or dry respirable particles have a mass average aerodynamic diameter (MMAD) of less than about 10 microns, preferably about 5 microns or less.

[0046] As used herein, the term “respiratory tract” includes the upper respiratory tract (e.g., nasal passages, nasal cavity, throat and pharynx), respiratory airways (e.g., larynx, trachea, bronchi and bronchioles) and lungs (e.g. respiratory bronchioles, alveolar ducts, alveolar sacs and alveoli).

[0047] The term “small” as used herein to describe respirable dry particles refers to particles that have a volume mean geometric diameter (VMGD) of about 10 microns or less, preferably about 5 microns or less.

Dry Powders and Dry Particles

[0048] The invention relates to respirable dry powder and respirable dry particles containing tiotropium as an active ingredient. The chemical structure of tiotropium was first described in US Patent. 5,610,163 and RE 39,820. Salts of tiotropium include salts containing cationic tiotropium with one of the following anions: bromide, fluoride, chloride, iodine, C1-C4-alkylsulfate, sulfate, hydrogensulfate, phosphate, hydrogenphosphate, dihydrogen phosphate, nitrate, maleate, acetate, trifluoroacetate, citrate, fumarate, tartrate, oxalate, succinate and benzoate, C1-C4-alkylsulfonate, which may be optionally mono-, dior tri-substituted by fluorine in the alkyl group, or phenylsulfonate which may optionally be mono- or poly-substituted by C1- C4-alkyl on the phenyl ring. Tiotropium bromide is an anticholinergic that provides therapeutic benefits, for example in the treatment of COPD and asthma, and is the active ingredient in SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) (Boehringer Ingelheim, Germany). Tiotropium bromide is known to crystallize in various forms, such as the crystalline anhydrous form (described, for example, in US Patents 6,608,055; 7,968,717; and 8,163,913 (Form 11)), crystalline monohydrate ( described, for example, in US Patents 6,777,423 and 6,908,928) and crystalline solvates (described, for example, in US Patent 7,879,871). The various crystalline forms of tiotropium can be distinguished by a number of different tests, including X-ray Powder Diffraction (XRPD), Differential Scanning Calorimetry (DSC), crystal structure, and infrared (IR) spectrum analysis. Tiotropium can be synthesized using a variety of methods that are well known in the art (including, for example, methods described in US Patents 6,486,321; 7,491,824; 7,662,963; and 8,344,143).

[0049] Tiotropium is generally present in respirable dry powders and respirable dry particles, in the form of tiotropium bromide. In particular, respirable dry powders comprise respirable dry particles that contain on a dry basis: from about 79.5% (w/w) to about 80.5% (w/w) sodium chloride, about 19. 5% (w/w) to about 20.5% (w/w) leucine and from about 0.01% (w/w) to about 0.5% (w/w) tiotropium bromide , and/or a weight ratio of sodium chloride to leucine of about 4:1, and about 0.01% (w/w) to about 0.5% (w/w) tiotropium bromide. For example, the invention provides respirable dry powders referred to as IV Formulations, listed in Table 1. Table 1. Tiotropium Formulations

[0050] Additionally, tiotropium formulations can be adjusted to allow for an additional therapeutic agent. In one embodiment, the respirable dry powders comprise respirable dry particles that contain on a dry basis: from about 60% to about 90% sodium chloride, about 10% to about 40% leucine, about 0.01 % to about 0.5% tiotropium bromide and, optionally, one or more additional therapeutic agents up to about 20%. In another embodiment, the respirable dry powders comprise respirable dry particles that contain on a dry basis: about 67% to about 84% sodium chloride, about 12% to about 28% leucine, about 0.01 to about 0.5% tiotropium bromide and, optionally, one or more additional therapeutic agents up to about 20%. Optionally, the sodium chloride content is from about 75% to about 82% and/or the leucine content is from about 15% to about 25%. Tiotropium bromide is preferably present in an amount of between about 0.02% and 0.25% by weight on a dry basis. The one or more additional therapeutic agents are preferably present in an amount of between about 0.01% to about 10%, more preferably between about 0.01% to 0.5%, greater than 0.5% to 3%. , or greater than 3% to about 10%. In another embodiment, the respirable dry powder comprising respirable dry particles containing, on a dry basis, about 65% to about 86% sodium chloride, about 10% to about 35% leucine, about 0. 01% to about 0.5% tiotropium bromide and, optionally, one or more additional therapeutic agents, wherein the one or more additional therapeutic agents is from about 1% to about 10%, more preferably, the one or more plus additional therapeutic agents is about 3% to about 7%, and more preferably, the one or more additional therapeutic agents is about 4% to 5%. The tiotropium bromide formulations described in this paragraph are collectively referred to as “Expanded Tiotropium Formulations”.

[0051] Additional preferred therapeutic combinations with tiotropium include corticosteroids, such as inhaled corticosteroids (ICS), long-acting beta agonists (LABA), short-acting beta agonists (SABA), anti-inflammatory agents and any combination thereof. A bifunctional beta2-agonist muscarinic antagonist (MABA) is optionally included among these additional therapeutic combinations. In a more preferred embodiment, tiotropium is combined with one or more ICS. Particularly preferred therapeutic combinations with tiotropium include: a) tiotropium and corticosteroids, such as inhaled corticosteroids (ICS); b) tiotropium and long-acting beta agonists (LABA); c) tiotropium and short-acting beta agonists (SABA); d) tiotropium and anti-inflammatory agents; e) tiotropium and MABA, f) tiotropium and a bronchodilator and any combination thereof. Combinations of these include, among others, tiotropium and ICS and LABA.

[0052] Suitable corticosteroids, such as inhaled corticosteroids (ICS), include budesonide, fluticasone, flunisolide, triamcinolone, beclomethasone, mometasone, ciclesonide, dexamethasone and the like.

[0053] Tiotropium can be delivered once daily (QD) to patients, so inhaled corticosteroids whose pharmacological data and dose regimen support once daily administration are preferred. Preferred inhaled corticosteroids are fluticasone, e.g. fluticasone furoate; mometasone, e.g. mometasone furoate; ciclesonide and the like.

[0054] Suitable LABAs include salmeterol, formoterol and isomers (for example arformoterol), clenbuterol, tulobuterol, vilanterol (Revolair™), indacaterol, carmoterol, isoproterenol, procaterol, uvaterol, milveterol, olodaterol and the like.

[0055] Suitable SABAs include albuterol, epinephrine, pirbuterol, levalbuterol, metaproteronol, maxair and the like.

[0056] Suitable MABAs include AZD 2115 (AstraZeneca), GSK961081 (GlaxoSmithKline), LAS190792 (Almirall), PF4348235 (Pfizer) and PF3429281 (Pfizer).

[0057] Combinations of corticosteroids and LABAs include salmeterol with fluticasone, formoterol with budesonide, formoterol with fluticasone, formoterol with mometasone, indacaterol with mometasone, and the like.

[0058] Suitable anti-inflammatory agents include leukotriene inhibitors, phosphodiesterase 4 (PDE4) inhibitors, other anti-inflammatory agents and the like.

[0059] Other suitable anti-inflammatory agents are kinase inhibitors.

[0060] Other anti-inflammatory agents include omalizumab (anti-IgE immunoglobulin, Daiichi Sankyo Company, Limited), Zolair (anti-IgE immunoglobulin, Genentech Inc, Novartis AG, Roche Holding Ltd), Solfa (LTD4 antagonist and phosphodiesterase inhibitor, Takeda Pharmaceutical Company Limited), IL-13 and IL-13 receptor inhibitors (such as AMG-317, MILR1444A, CAT-354, QAX576, IMA-638, Anrukinzumab, IMA-026, MK-6105, DOM-0910 and similar), IL-4 and IL-4 receptor inhibitors (such as Pitrakinra, AER-003, AIR-645, APG-201, DOM-0919 and the like), IL-1 inhibitors such as canakinumab, receptor antagonists of CRTh2 such as AZD1981 (CRTh2 receptor antagonists, AstraZeneca), neutrophil elastase inhibitor such as AZD9668 (AstraZeneca neutrophil elastase inhibitors), P38 mitogen-activated protein kinase inhibitor, e.g. GW856553X Losmapimod, GSK681323, GSK 856553 and GSK610677 (all P38 kinase inhibitors, GlaxoSmithKline PLC) and PH797804 (P38 kinase inhibitors; Pfizer), Arofylline LAB ALMIRALL (PDE-4 inhibitor, Laboratorios Almirall, S.A.), ABT761 (5-LO inhibitor, Abbott Laboratories), Zyflo® (5-LO inhibitor, Abbott Laboratories), BT061 (anti-CD4 mAb, Boehringer Ingelheim GmbH), BIBW 2948 BS (map kinase inhibitor), Corus (inhaled lidocaine to decrease eosinophils, Gilead Sciences Inc.), Prograf® (IL-2-mediated T cell activation inhibitor, Astellas Pharma), Bimosiamose PFIZER INC (selectin inhibitor, Pfizer Inc), R411 (alpha4beta1/alpha4beta7 integrin antagonist, Roche Holdings Ltd), Tilade® (inflammatory mediator inhibitor, Sanofi-Aventis), Orenica® (T cell co-stimulation inhibitor, Bristol -Myers Squibb Company), Soliris® (anti-C5, Alexion Pharmaceuticals Inc), Entorken® (Farmacija d.o.o.), Excellair® (Syk kinase siRNA, ZaBeCor Pharmaceuticals, Baxter International Inc), KB003 (anti-GMCSF mAb, KaloBios Pharmaceuticals) , Cromolyn sodium (mast cell mediator release inhibitor): Cromolyn sodium BOEHRINGER (Boehringer Ingelheim GmbH), Cromolyn sodium TEVA (Teva Pharmaceutical Industries Ltd), Intal (Sanofi-Aventis), BI1744CL (oldaterol (beta-2 adrenergic receptor antagonist ) and tiotropium, Boehringer Ingelheim GmbH), NFkappa-B inhibitors, CXR2 antagonist, HLE inhibitor, HMG-CoA reductase inhibitor and the like.

[0061] Anti-inflammatory agents also include compounds that inhibit/decrease cell signaling by inflammatory molecules such as cytokines (e.g., IL-1, IL-4, IL-5, IL-6, IL-9, IL-13, IL-18, IL-25, IFN-α, IFN-β and others), CC chemokine CCL-1 - CCL28 (some of which are also known as, for example, MCP-1, CCL2, RANTES), CXC chemokines CXCL1 - CXCL17 (some of which are also known as, for example, IL-8, MIP-2), CXCR2, growth factors (for example, GM-CSF, NGF, SCF, TGF-β, EGF, VEGF and others) and/or their respective receivers.

[0062] Some of the examples of the above-mentioned anti-inflammatory antagonists/inhibitors include ABN912 (MCP-1/CCL2, Novartis AG), AMG761 (CCR4, Amgen Inc), Enbrel® (TNF, Amgen Inc, Wyeth), huMAb OX40L GENENTECH (TNF superfamily, Genentech Inc, AstraZeneca PLC), R4930 (TNF superfamily, Roche Holding Ltd), SB683699/Firategrast (VLA4, GlaxoSmithKline PLC), CNT0148 (TNFalfa, Centocor, Inc, Johnson & Johnson, Schering-Plough Corp) ; Canakinumab (IL-beta, Novartis); Israpafant MITSUBISHI (PAF/IL-5, Mitsubishi Tanabe Pharma Corporation); IL-4 and IL-4 receptor antagonists/inhibitors: AMG317 (Amgen Inc), BAY169996 (Bayer AG), AER-003 (Aerovance), APG-201 (Apogenix); IL-5 and IL-5 receptor antagonists/inhibitors: MEDI563 (AstraZeneca PLC, MedImmune, Inc), Bosatria® (GlaxoSmithKline PLC), Cinquil® (Ception Therapeutic), TMC120B (Mitsubishi Tanabe Pharma Corporation), Bosatria (GlaxoSmithKline PLC ), Reslizumab SCHERING (Schering-Plough Corp); MEDI528 (IL-9, AstraZeneca, MedImmune, Inc); IL-13 and IL-13 receptor antagonists/inhibitors: TNX650 GENENTECH (Genentech), CAT-354 (AstraZeneca PLC, MedImmune), AMG-317 (Takeda Pharmaceutical Company Limited), MK6105 (Merck & Co Inc), IMA- 026 (Wyeth), IMA-638 Anrukinzumab (Wyeth), MILR1444A/Lebrikizumab (Genentech), QAX576 (Novartis), CNTO-607 (Centocor), MK-6105 (Merck, CSL); Dual inhibitors of IL-4 and IL-13: AIR645/ISIS369645 (ISIS Altair), DOM-0910 (GlaxoSmithKline, Domantis), Pitrakinra/AER001/Aerovant™ (Aerovance Inc), AMG-317 (Amgen) and the like. CXCR2 antagonists include, for example, Reparixin (Dompe S.P.A.), DF2162 (Dompe, S.P.A.), AZ-10397767 (AstraZeneca), SB656933 (GlaxoSmithKline PLC), SB332235 (GlaxoSmithKline PLC), SB468477 (GlaxoSmithKline PLC), and SCH5271 23 (Schering- Plow Corp).

[0063] In Formulations I to V and/or Expanded Tiotropium Formulations, the respirable dry powders and/or respirable dry particles are preferably small, dense in mass, and dispersible. To measure the geometric volume mean diameter (VMGD), the laser diffraction system can be used, for example, a Spraytec system (particle size analysis instrument, Malvern Instruments) and a HELOS/RODOS system (diffraction sensor laser with dry dispensing unit, Sympatec GmbH). The respirable dry particles of Formulations I through V have a VMGD as measured by laser diffraction at the 1.0 bar dispersion pressure setting using the HELOS/RODOS system of about 10 microns or less (e.g., about 0. 5 μm to about 10 μm), about 5 microns or less (e.g., about 0.5 μm to about 5 μm), about 4 μm or less (e.g., about 0.5 μm to about 4 μm), about 3 μm or less (e.g., about 0.5 μm to about 3 μm), about 1 μm to about 5 μm, about 1 μm to about 4 μm, about 1.5μm to about 3.5μm, about 2μm to about 5μm, about 2μm to about 4μm, or about 2μm to about 3μm. Preferably, the VMGD is about 5 microns or less (e.g., about 1 μm to about 5 μm), or about 4 μm or less (e.g., about 1 μm to about 4 μm).

[0064] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or Expanded Tiotropium Formulations are dispersible, and have a ratio of 1 bar/4 bar and/or 0.5 bar/4 bar of less than about 2.0 (e.g., about 0.9 to less than about 2), about 1.7 or less (e.g., about 0.9 to about 1.7) about 1 .5 or less (e.g., about 0.9 to about 1.5), about 1.4 or less (e.g., about 0.9 to about 1.4), or about 1. 3 or less (e.g., about 0.9 to about 1.3), and preferably has 1 bar/4 bar and/or 0.5 bar/4 bar of about 1.5 or less (e.g., about 1.0 to about 1.5), and/or from about 1.4 or less (e.g., about 1.0 to about 1.4).

[0065] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations preferably have a compacted density of at least about 0.45 g/cm3 (e.g., about 0. 45 g/cm3 to about 1.2 g/cm3), at least about 0.5 g/cm3 (e.g., about 0.5 g/cm3 to about 1.2 g/cm3), of at least at least about 0.55 g/cm3 (e.g., about 0.55 g/cm3 to about 1.2 g/cm3), at least about 0.6 g/cm3 (e.g., about 0 .6 g/cm3 to about 1.2 g/cm3), or from at least about 0.6 g/cm3 to about 1.0 g/cm3.

[0066] Furthermore, the respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations may have a compact density of at least about 0.4 g/cm3, a greater compact density than 0.4 g/cm3, or a compacted density greater than 0.4 g/cm3 to about 1.2 g/cm3.

[0067] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or Expanded Tiotropium Formulations have an MMAD of less than 10 microns (e.g., about 0.5 microns less than 10 microns) , preferably an MMAD of about 5 microns or less (e.g., about 1 micron to about 5 microns), about 2 microns to about 5 microns, or about 2.5 microns to about 4.5 microns . In a preferred embodiment, MMAD is measured using a capsule-based passive dry powder inhaler (RS01 Model 7, High resistance Plastiape S.p.A.), which has a specific resistance of 0.036 sqrt(kPa)/liters per minute, and as measured at 60 LPM, the preferred MMAD range is about 2.9 microns to about 4.0 microns, and the most preferred MMAD range is about 2.9 microns to about 3.5 microns.

[0068] In another preferred embodiment, MMAD is measured using an RS01 UHR2 capsule-based passive dry powder inhaler (RS01 Model 7, Ultrahigh resistance 2 (UHR2) Plastiape S.p.A.), which has a specific resistance of 0.048 sqrt( kPa)/liters per minute, and as measured at 39 LPM, the preferred MMAD range is about 3.0 microns to about 5.0 microns, and the most preferred MMAD range is about 3.8 microns to about 4.3 microns.

[0069] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations have an FPF of less than about 5.6 microns (FPF<5.6 μm) of a total dose of at least about 35%, preferably of at least about 45%, of at least about 60%, between about 45% and about 80%, or between about 60% and about 80%.

[0070] Furthermore, the respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations preferably have an FPF of less than about 3.4 microns (FPF<3.4 μm) of a total dose of at least about 20%, preferably at least about 25%, at least about 30%, at least about 40%, between about 25% and about 60%, or between about 40 % and about 60%.

[0071] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations have an FPD of less than about 4.4 microns (FPD<4.4 μm) of about 1 microgram and about 5 micrograms, or about 2.0 micrograms and about 5.0 micrograms.

[0072] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations have an FPD of less than about 4.4 microns (FPD<4.4 μm), as a percentage of total dose of at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55% or at least 60%.

[0073] In some aspects, the invention provides an effective method of dose delivery of tiotropium as a dry powder. The release efficiency of a dose of tiotropium can be characterized based on the delivery of an effective amount of tiotropium to the lungs at a lower nominal dose introduced into the capsule than from a standard dry powder formulation such as SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) which has a nominal dose of 18 micrograms tiotropium. The release efficiency of a dose of tiotropium can be further characterized by the release of a fine particle dose similar to that of a SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) capsule with a lower nominal dose introduced into the capsule. The release efficiency of a dose of tiotropium can be further characterized by the release of a fine particle dose of less than about 4.4 microns (FPD<4.4 μm) similar to that of a SPIRIVA (tiotropium bromide) capsule. ) HANDIHALER (dry powder inhaler) with a lower nominal dose introduced into the capsule.

[0074] The delivery efficiency of a dose of tiotropium can be further characterized in one aspect of the current invention, based on delivering an effective amount of tiotropium to the lungs to achieve a similar improvement in lung function, preferably, a similar change in forced expiratory volume in one second (FEV1), or more preferably, a similar change in steady-state FEV1 response as SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), but with a smaller nominal dose than SPIRIVA (bromide of tiotropium) HANDIHALER (dry powder inhaler). In one aspect, when measured in patients who are being administered the respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations of the present invention; when the nominal dose of tiotropium in the dry respirable powders and/or dry respirable particles of Formulations I to V and/or the Expanded Tiotropium Formulations is 70% or less, 50% or less, or preferably 35% or less, 25% or less, or 20% or less, 15% or less, 10% or less, or 5% or less than the nominal dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which is 18 micrograms tiotropium ; the change in FEV1 is about 80% or greater than the change in FEV1 observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), preferably about 85% or greater than the change in FEV1 observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), more preferably, 90% or greater than the change in FEV1 observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), or more preferably, about 95% or greater than the change in FEV1 observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler).

[0075] In another aspect, when measured in patients who are receiving the respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations of the present invention; when the nominal dose of tiotropium in the dry respirable powders and/or dry respirable particles of Formulations I to V and/or the Expanded Tiotropium Formulations is 70% or less, 50% or less, or preferably 35% or less, 25% or less; or 20% or less, 15% or less, 10% or less, or 5% or less than the nominal dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which is 18 micrograms tiotropium; the change in steady-state FEV1 response is about 80% or more than the change in steady-state FEV1 response observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), preferably about of 85% or more than the change in steady-state FEV1 response observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), more preferably, 90% or more than the change in FEV1 response at steady state observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), or more preferably, about 95% or more than the change in steady-state FEV1 response observed in patients taking SPIRIVA ( tiotropium bromide) HANDIHALER (dry powder inhaler).

[0076] In another aspect, when measured in patients who are receiving the respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations of the present invention; when the nominal dose of tiotropium in the dry respirable powders and/or dry respirable particles of Formulations I to V and/or the Expanded Tiotropium Formulations is 70% or less, 50% or less, or preferably 35% or less, 25% or less; or 20% or less, 15% or less, 10% or less, or 5% or less than the nominal dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which is 18 micrograms tiotropium; the change in FEV1 response at steady state is about 80 mL or more, about 90 mL or more, preferably about 100 mL or more, about 110 mL or more, about 120 mL or more.

[0077] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations may be contained in a container that may contain about 15 mg, 10 mg, 7.5 mg, 5 mg , 2.5 mg, or 1 mg of respirable dry powder mass. Such containers may contain a nominal dose of tiotropium ranging from about 3 to about 12 micrograms, from about 3 to about 9 micrograms, or from about 3 to about 6 micrograms. In certain embodiments, the container may contain a nominal dose of tiotropium of about 3 micrograms, about 6 micrograms, about 9 micrograms, or about 12 micrograms. The container may be contained in a dry powder inhaler or may be packaged and/or sold separately.

[0078] Furthermore, the containers may contain a nominal dose of tiotropium ranging from about 1.5 micrograms to 12 micrograms. In certain embodiments, the container may contain a nominal dose of tiotropium of about 2 micrograms.

[0079] Additionally, the containers may contain a nominal dose of tiotropium ranging from about 0.5 micrograms to about 6 micrograms, or from about 0.5 micrograms to about 3 micrograms, or from about 1 microgram to about 3 micrograms. In a certain embodiment, the container may contain a nominal dose of tiotropium of about 0.5 micrograms, about 1 microgram, about 1.5 micrograms, or about 2.5 micrograms.

[0080] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or Expanded Tiotropium Formulations may have a water and solvent content of up to about 15% by weight of respirable dry powder or particle. For example, the water or solvent content is up to about 10%, up to about 5%, or preferably between about 0.1% and about 3%, from about 0.01% to 1%, or they may be substantially free of water or other solvent or be anhydrous.

[0081] Respirable dry powders and/or respirable dry particles of Formulations I to V and/or Expanded Tiotropium Formulations can be administered with low inhalation energy. In order to relate powder dispersion to different inhalation flow rates, volumes and from inhalers of different resistance, the energy required to perform the inhalation maneuver can be calculated. Inhalation energy can be calculated from the equation E=R2Q2V where E is the inhalation energy in joules, R is the inhaler resistance in kPa1/2/LPM, Q is the constant flow rate in L/min and V is the volume of air inhaled in L.

[0082] The respirable dry powders and/or respirable dry particles of Formulations I to V and/or Expanded Tiotropium Formulations are characterized by a high emitted dose (e.g., CEPM of at least 75%, at least 80%, at least at least 85%, at least 90%, at least 95%) from a dry powder inhaler when the total inhalation energy is less than 5 joules, less than about 3.5 joules, less than about 2.4 joules, less than about 2 joules, less than about 1 joule, less than about 0.8 joule, less than about 0.5 joule, or less than about 0.3 joule is applied to the dry powder inhaler. For example, an emitted dose of at least 75%, at least 80%, at least 85%, at least 90%, at least 95% of the CEPM of any of Formulations I through V contained in a unit dose container, containing about 4 mg or more, about 6 mg or more, about 11 mg or more, about 15 mg or more, about 20 mg or more, about 30 mg or more, or about 4 mg to about 6 mg, about 6 mg to about 11 mg, about 11 mg to about 15 mg, or about 15 mg to about 20 mg of the appropriate formulation in a dry powder inhaler can be achieved when full energy inhalation rate of less than about 5 joules, less than about 3.5 joules, less than about 2 joules, less than about 1 joule, and less than about 0.8 joules, less than about of 0.5 joule, or less than about 0.3 joule is applied to the dry powder inhaler.

[0083] In one aspect, the respirable dry powders and/or respirable dry particles of Formulations I to V and/or the Expanded Tiotropium Formulations are characterized by a powder mass emitted from the capsule of at least 80% when emitted from an inhaler of passive dry powder that has a resistance of about 0.036 sqrt(kPa)/liters per minute under the following conditions: an inhalation energy of 2.3 joules at a flow rate of 30 LPM using a size 3 capsule, which contains a total mass of 10 mg, said total mass consisting of dry respirable particles of any of Formulations I to V, wherein the volume-average geometric diameter of dry respirable particles emitted from the inhaler is 5 microns or less.

[0084] The dry powder may fill the unit dose container, or the unit dose container may be at least 2% full, at least 5% full, at least 10% full, at least 20% full, at least 30% full, at least 40% full, at least 50% full, at least 60% full, at least 70% full, at least 80% full, or at least 90% full. The unit dose container may be a capsule (e.g., size 000, 00, 0E, 0, 1, 2, 3, and 4, with respective volumetric capacities of 1.37 ml, 950 μl, 770 μl, 680 μl, 480 μl, 360 μl, 270 μl and 200 μl). The capsule is preferably between about 2% full and about 10% full, between about 10% full and about 20% full. The unit dose container may be a blister. The blister may be packaged as a single blister, or as part of a set of blisters, for example, 7 blisters, 14 blisters, 28 blisters, or 30 blisters. One or more blisters is preferably at least 30% full, at least 50% full, or at least 70% full.

[0085] Healthy adult populations are predicted to be able to achieve inhalation energies ranging from 2.9 joules for comfortable inhalations to 22 joules for maximal inhalations using peak inspiratory flow rate (PIFR) values measured by Clarke et al. (Journal of Aerosol Med, 6(2), p.99-110, 1993) for the flow rate Q of two inhaler resistances of 0.02 and 0.055 kPa1/2/LPM, with an inhalation volume of 2L based on both the FDA's guidance for dry powder inhalers and the work of Tiddens et al. (Journal of Aerosol Med, 19(4), p.456-465, 2006) which found adults with an average inhaled volume of 2.2L across a variety of DPIs.

[0086] Adult patients with mild, moderate and severe COPD are expected to be able to achieve maximum inhalation energy of 5.1 to 21 Joules, 5.2 to 19 Joules and 2.3 to 18 Joules, respectively. This is again based on using the measured PIFR values for flow rate Q in the equation for inhalation energy. The achievable PIFR for each group is a function of the resistance of the inhaler being inhaled. The work of Broeders et al. (Eur Respir J, 18, p.780-783, 2001) was used to predict the maximum and minimum PIFR achievable through 2 dry powder inhalers of resistances of 0.021 and 0.032 kPa1/2/LPM for each.

[0087] Likewise, it is predicted that adult asthmatic patients will be able to achieve maximum inhalation energy of 7.4 to 21 Joules based on the same assumptions as the COPD population and PIFR data from Broeders et al.

[0088] Healthy adults and children, patients with COPD, asthmatic patients aged 5 years and older, and patients with CF, for example, are capable of providing sufficient inhalation energy to deflate and disperse the dry powder formulations of the invention.

[0089] An advantage of the invention is the production of powders that disperse well over a wide flow range and are relatively independent of flow. The respirable dry particles and respirable dry powders of the invention enable the use of simple, passive DPI for a large patient population.

[0090] In some aspects, the invention is a respirable dry powder containing respirable dry particles of any one of Formulations I to V. In another particular aspect, the respirable dry powder containing respirable dry particles comprises i) about 79.5% to about 80.5% sodium chloride, ii) about 19.5% to about 20.5% leucine and iii) about 0.01% to about 0.5% tiotropium bromide, where all values are percentages by weight. In a further particular aspect, the invention is a respirable dry powder containing respirable dry particles containing Expanded Tiotropium Formulations.

[0091] Respirable dry powder containing respirable dry particles of any of the formulations of the previous paragraph or being in any of the ranges of the formulation of the previous paragraph is characterized by: 1. VMGD at 1 bar, as measured using a HELOS/system WHEELS, is about 10 microns or less, preferably between about 1 micron and about 5 microns, between about 1 micron and about 4.0 microns, or between about 1.5 microns and about 3.5 microns ; 2. 1 bar/4 bar of about 1.5 or less, about 1.4 or less, or between about 1 and about 1.5, or between about 1 and about 1.4; 3. 0.5 bar/4 bar of about 1.5 or less, about 1.4 or less, or between about 1 and about 1.5, or between about 1 and about 1.4; 4. compacted density of about 0.45 g/cm3 or more, between about 0.45 g/cm3 and about 1.2 g/cm3, between 0.5 g/cm3 and about 1.2 g/cm3 cm3, between 0.55 g/cm3 and about 1.1 g/cm3, or between 0.6 g/cm3 and about 1 g/cm3; 5. MMAD of about 10 microns or less, preferably between about 1 micron and about 5 microns, or between about 2.5 microns and about 4.5 microns; 6. FPF<5.6 of at least about 45%, at least about 60%, or between about 60% and about 80%; 7. FPF<4.4 of at least about 35%, at least about 50%, or between about 50% and about 70%; 8. FPF<3.4 of at least about 25%, at least about 40%, or between about 40% and about 60%; 9. FPD<4.4 of between about 1 microgram and about 5 micrograms tiotropium, or between about 2.0 micrograms and about 5.0 micrograms tiotropium; and/or 10. Ratio of fine particle dose less than 2.0 microns to fine particle dose less than 4.4 microns of less than about 0.50, less than about 0.35, or less than about 0.25.

[0092] The respirable dry powder or respirable dry particles described above may further be characterized by water content of less than 15% by weight, preferably less than 10%, less than 5%, or more preferably less than 1%, all by weight . Furthermore, the respirable dry powder or respirable dry particles of any of Formulations I to V are characterized by a mass of dust emitted from the capsule of at least 80% when emitted from a passive dry powder inhaler having a resistance of about 0.036 sqrt(kPa)/liters per minute under the following conditions: an inhalation energy of 2.3 joules at a flow rate of 30 LPM using a size 3 capsule, which contains a total mass of 10 mg, said mass total consists of dry respirable particles of any of Formulations I to V, wherein the volume-averaged geometric diameter of dry respirable particles emitted from the inhaler is 5 microns or less.

[0093] In another particular aspect, the invention is a respirable dry powder containing respirable dry particles of a formulation falling within one of the following ranges or specifically identified formulation: about 60% to about 90% sodium chloride, about 10% to about 40% leucine, about 0.01% to about 0.5% tiotropium, and optionally one or more additional therapeutic agents up to about 20%; about 65% to about 86% sodium chloride, about 10% to about 35% leucine, about 0.01% to about 0.5% tiotropium, and optionally one or more additional therapeutic agents up to about 20%; preferably, about 67% to about 84% sodium chloride, about 12% to about 28% leucine, about 0.01 to about 0.5% tiotropium, and optionally one or more therapeutic agents additional amounts of up to around 20%; more preferably, about 75% to about 82% sodium chloride, about 15% to about 25% leucine, about 0.01 to about 0.5% tiotropium, and optionally one or more agents additional therapeutics up to approximately 20%; or more preferably, about 79.5% to about 80.5% sodium chloride, about 19.5% to about 20.5% leucine, about 0.01 to about 0.5% of tiotropium; or IV Formulations, where the weight percentage of each component is listed below in Table 1A: Table 1A. Tiotropium formulations where all values are percentage by weight and all components of any formulation quantify 100%; wherein the tiotropium is preferably present in an amount between about 0.02% and 0.25%, by weight on a dry basis, and wherein the one or more additional therapeutic agents is preferably present in an amount between about 0. .01% and about 10%. Preferably, the additional therapeutic agent is an inhaled corticosteroid (ICS). Most preferably, the ICS is chosen to combine with the tiotropium dose regimen, which is administered once daily (QD), for example, as indicated in the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) leaflet. Preferred inhaled corticosteroids are fluticasone, such as fluticasone furoate, mometasone, such as mometasone furoate; ciclesonide and the like; and the most preferred inhaled corticosteroids are fluticasone furoate, mometasone furoate, or ciclesonide. The preferred weight percentage on a dry basis of these inhaled corticosteroids is about 0.2% to about 4% for fluticasone furoate, about 0.4% to about 10% for mometasone furoate, and about 0.2% to about 5% for ciclesonide. In a preferred aspect, only one ICS is used in the formulation. In one aspect, the weight ratio of sodium chloride to leucine is about 4:1. In one aspect, the respirable dry powder consists of respirable dry particles. In another aspect, the formulation does not contain any therapeutic agent other than tiotropium.

[0094] Respirable dry powder, comprising respirable dry particles, described by any of the ranges or formulations specifically disclosed in the previous paragraph is characterized by: 1. VMGD at 1 bar, as measured using a HELOS/RODOS system, is about of 10 microns or less, preferably between about 1 micron and about 5 microns, between about 1.0 microns and about 4.0 microns, or between about 1.5 microns and about 3.5 microns, or between about 2 microns and 5 microns, or between 2.5 microns and 4.5 microns; 2. 1 bar/4 bar of about 1.5 or less, about 1.4 or less, about 1.3 or less, or between about 1 and about 1.5, or between about 1 and about 1.4, or between about 1 and about 1.3; 3. 0.5 bar/4 bar of about 1.5 or less, about 1.4 or less, about 1.3 or less, or between about 1 and about 1.5, or between about 1 and about 1.4, or between about 1 and about 1.3; 4. Compacted density greater than 0.4 g/cm3, greater than 0.4 g/cm3 to about 1.2 g/cm3, about 0.45 g/cm3 or more, between about 0.45 g/cm3 and about 1.2 g/cm3, between 0.5 g/cm3 and about 1.2 g/cm3, between 0.55 g/cm3 and about 1.1 g/cm3, or between 0 .6 g/cm3 and about 1 g/cm3; 5. MMAD of about 10 microns or less, preferably between about 1 micron and about 5 microns, between about 2.5 microns and about 4.5 microns, or between about 3.0 microns and about 5 .0 microns; 6. FPF<5.0 of at least about 45%, at least about 60%, or between about 60% and about 80%; 7. FPF<4.4 of at least about 35%, at least about 50%, or between about 50% and about 70%; 8. FPF<3.4 of at least about 25%, at least about 40%, or between about 40% and about 60%; 9. FPD<4.4 between about 1 microgram and about 5 micrograms tiotropium, between about 2.0 micrograms and about 5.0 micrograms tiotropium; or, preferably, between 2.0 micrograms and 4.0 micrograms; 10. FPD<5.0 of between about 1 microgram and about 5 micrograms tiotropium, between about 2.0 micrograms and about 5.0 micrograms tiotropium; or, preferably, between 2.0 micrograms and 4.0 micrograms; 11. Ratio of fine particle dose less than 2.0 microns to fine particle dose less than 4.4 microns of less than about 0.50, less than about 0.35, less than about of 0.30; preferably less than about 0.25, less than about 0.20, less than about 0.18, or about 0.15 or less; 12. Ratio of fine particle dose less than 2.0 microns to fine particle dose less than 5.0 microns of less than about 0.50, less than about 0.35, less than about 0.30; preferably less than about 0.25, less than about 0.20, less than about 0.18, or about 0.15 or less; 13. Water content, on a dry basis, of less than 15%, less than 10%, preferably less than 5%, or more preferably less than 1%; 14. A CEPM of at least 80% when emitted from a passive dry powder inhaler that has a resistance of about 0.036 sqrt(kPa)/liters per minute under the following conditions: an inhalation energy of 2.3 Joules at a flow rate of 30 LPM using a size 3 capsule, which contains a total mass of 10 mg of dry respirable powder comprising dry respirable particles described by any of the ranges or formulations specifically disclosed in the preceding paragraph, wherein the VMGD of dry particles respirable emitted from the inhaler is 5 microns or less; 15. The emitted dose of at least 70%, at least 75%, at least 80%, or at least 85% when emitted from a passive dry powder inhaler that has a resistance of about 0.036 sqrt(kPa)/ liters per minute under the following conditions: an inhalation energy of 2.3 Joules at a flow rate of 30 LPM using a size 3 capsule, which contains a total mass of 10 mg of respirable dry powder comprising respirable dry particles described by any of the ranges or formulations specifically disclosed in the preceding paragraph, wherein the VMGD of respirable dry particles emitted from the inhaler is 5 microns or less; 16. A CEPM of at least 80% when emitted from a passive dry powder inhaler that has a resistance of about 0.048 sqrt(kPa)/liters per minute under the following conditions: an inhalation energy of 1.8 Joules at a flow rate of 20 LPM using a size 3 capsule, which contains a total mass of 5 mg of dry respirable powder comprising dry respirable particles described by any of the ranges or formulations specifically disclosed in the preceding paragraph, wherein the VMGD of dry particles respirable emitted from the inhaler is 5 microns or less; and/or 17. The emitted dose of at least 65%, at least 70%, at least 75%, at least 80%, or at least 85% when emitted from a passive dry powder inhaler that has a resistance of about 0.048 sqrt(kPa)/liters per minute under the following conditions: an inhalation energy of 1.8 Joules at a flow rate of 20 LPM using a size 3 capsule, which contains a total mass of 5 mg of dry respirable powder comprising respirable dry particles described by any of the ranges or formulations specifically disclosed in the preceding paragraph, wherein the VMGD of respirable dry particles emitted from the inhaler is 5 microns or less.

[0095] The respirable dry powder comprising respirable dry particles described by any of the specifically disclosed ranges or formulations characterized in the previous paragraph may be packaged in a container, for example, a capsule or a blister. When the container is a capsule, the capsule is, for example, a size 2 or 3 capsule, and is preferably a size 3 capsule. The capsule material can be, for example, gelatin or HPMC (hydroxypropylmethylcellulose), and more preferably HPMC.

[0096] The respirable dry powder comprising respirable dry particles described and characterized above can be packaged in a dry powder inhaler (DPI). The DPI may be a capsule-based DPI or a blister-based DPI and is preferably a capsule-based DPI. More preferably, the dry powder inhaler is selected from the RS01 family (Plastiape S.p.A., Italy). More preferably, the dry powder inhaler is selected from the RS01 HR or RS01 UHR2. Most preferably, the dry powder inhaler is the RS01 UHR2. In one respect, the dry powder inhaler is not the RS01 HR.

[0097] When the respirable dry powder, comprising respirable dry particles, described and characterized above is contained in a container, the container can contain a mass of respirable dry powder comprising respirable dry particles between about 8 mg and about 12 mg, between between about 5.5 mg and about 9.5 mg, between about 3.5 mg and about 6.5 mg, between about 1.5 mg and 4.5 mg, or between 0.5 mg and 2 .5 mg; and preferably between about 3.5 mg and about 6.5 mg; or about 15 milligrams, about 10 milligrams, about 7.5 milligrams, about 5 milligrams, about 2.5 milligrams, or about 1 milligram; preferably a mass of about 5 milligrams. Alternatively or additionally, when respirable dry powder comprising respirable dry particles is contained in a container, the container may contain a nominal dose of tiotropium between about 1.5 micrograms to about 12 micrograms, between about 3 to about 12 micrograms. , between about 3 to about 6 micrograms; preferably, from about 0.5 micrograms to about 6 micrograms, or from about 0.5 micrograms to about 3 micrograms, or from about 1 microgram to about 3 micrograms; or a nominal dose of tiotropium of about 12 micrograms, about 9 micrograms, preferably about 6 micrograms, about 4 micrograms; or, more preferably, about 3 micrograms, about 2 micrograms or about 1 microgram.

[0098] When the respirable dry powder comprising respirable dry particles described and characterized above is administered to a patient; when the nominal dose of tiotropium is 70% or less, 50% or less, or preferably, 35% or less, 25% or less; or 20% or less, 15% or less, 10% or less, or 5% or less of a nominal dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which is 18 micrograms of tiotropium; the change in steady-state FEV1 response is about 80% or greater than the change in steady-state FEV1 response observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), preferably, about 85% or greater than the change in steady-state FEV1 response observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), more preferably, 90% or greater than the change in response of Steady-state FEV1 observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), or more preferably, about 95% or greater than the change in steady-state FEV1 response observed in patients taking SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Alternatively or in addition when the respirable dry powder, comprising respirable dry particles, described and characterized above is administered to a patient; when the nominal dose of tiotropium is 70% or less, 50% or less, or preferably, 35% or less, 25% or less; or 20% or less, 15% or less, 10% or less, or 5% or less of a nominal dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which is 18 micrograms of tiotropium; the change in FEV1 response at steady state is about 80 mL, about 90 mL or greater, preferably about 100 mL or greater, about 110 mL or greater, about 120 mL or greater. The patient described above may be a patient with a respiratory disease such as COPD, chronic bronchitis, emphysema, asthma, cystic fibrosis or non-cystic fibrosis bronchiectasis. The respiratory disease is preferably COPD, chronic bronchitis, and/or emphysema. The respirable dry powder comprising respirable dry particles described and characterized above may be administered to a patient to reduce the incidence or severity of an acute exacerbation of a respiratory disease, to alleviate the symptoms of a respiratory disease, and/or to improve pulmonary function of a patient with respiratory disease.

Methods for preparing dry powders and dry particles

[0099] Dry particles and respirable dry powders can be prepared using any suitable method. Many methods suitable for preparing respirable dry powders and particles are conventional in the art, and include single and double emulsion with solvent evaporation, spray drying, freeze drying, milling (e.g., jet mill), mixing, solvent extraction. , solvent evaporation, phase separation, simple and complex coacervation, interfacial polymerization, suitable methods involving the use of supercritical carbon dioxide (CO2), ultrasonically assisted crystallization, formation of nanoparticle aggregates and other suitable methods, including combinations thereof . Respirable dry particles can be made using methods for preparing microspheres or microcapsules known in the art. These methods can be employed under conditions that result in the formation of dry respirable particles with desired aerodynamic properties (e.g., aerodynamic diameter and geometric diameter). If desired, dry respirable particles with desired properties, such as size and density, can be selected using suitable methods, such as sieving.

[0100] Suitable methods for selecting respirable dry particles with desired properties, such as size and density, include wet sieving, dry sieving and aerodynamic classifiers (such as cyclones).

[0101] The dry respirable particles are preferably spray dried. Suitable spray drying techniques are described, for example, by K. Masters in “Spray Drying Handbook”, John Wiley & Sons, New York (1984). Generally, during spray drying, heating from a heated gas, such as heated air or nitrogen, is used to evaporate a solvent from droplets formed by atomization of a continuous liquid feed. When hot air is used, the moisture in the air is at least partially removed before use. When nitrogen is used, the nitrogen gas can be run “dry,” meaning no additional water vapor is combined with the gas. If desired, the moisture level of the nitrogen or air can be adjusted before spray drying operation begins to a fixed value above the “dry” nitrogen. If desired, spray drying or other instruments, e.g., jet mill instrument, used to prepare dry particles may include an in-line geometric particle sizer that determines the geometric diameter of the respirable dry particles as they are produced, and /or an in-line aerodynamic particle sizer that determines the aerodynamic diameter of dry respirable particles as they are produced.

[0102] For spray drying, solutions, emulsions or suspensions containing the dry particle components to be produced in a suitable solvent (e.g., aqueous solvent, organic solvent, aqueous-organic mixture or emulsion) are distributed into a container drying using an atomization device. For example, a nozzle or a rotating atomizer can be used to distribute the solution or suspension to the drying vessel. The nozzle can be a two-fluid nozzle, which is an internal mixing configuration or an external mixing configuration. Alternatively, a rotary atomizer with a 4- or 24-blade wheel can be used. Examples of suitable spray dryers that can be equipped with a rotary atomizer or nozzle include a Mobile Minor spray dryer or the model PSD-1, both manufactured by GEA Niro, Inc. (Denmark). Actual spray drying conditions will vary depending, in part, on the composition of the solution or suspension used in the spray drying process and the flow rate of the material. One skilled in the art will be able to determine appropriate conditions based on the compositions of the solution, emulsion or suspension to be spray dried, the desired properties of the particles and other factors. In general, the inlet temperature to the spray dryer is from about 90°C to about 300°C, and preferably is from about 220°C to about 285°C. Another preferred range is between 130°C to about 200°C. The spray dryer outlet temperature will vary depending on factors such as feed temperature and the properties of the materials being dried. Generally, the outlet temperature is from about 50°C to about 150°C, preferably from about 90°C to about 120°C, or about 98°C to about 108°C. Another preferred range is between 65°C to about 110°C, preferably about 75°C to about 100°C. If desired, the dry respirable particles that are produced can be fractionated by volumetric size, for example, using a sieve, or fractionated by aerodynamic size, for example, using the cyclone, and/or further separated according to density using known techniques for experts in the technique.

[0103] Other examples of spray dryers include the ProCepT Formatrix R&D spray dryer (ProCepT nv, Zelzate, Belgium). BüCHI B-290 MINI SPRAY DRYER (BÜCHI Labortechnik AG, Flawil, Switzerland). An additional preferred range for the inlet temperature to the spray dryer is from about 180°C to about 285°C. An additional preferred range for the outlet temperature for the spray dryer is from about 40°C to about 110°C, more preferably, from about 50°C to about 90°C.

[0104] To prepare the dry respirable particles of the invention, generally, a solution, emulsion or suspension containing desired components of the dry powder (i.e., a feed stock) is prepared and spray-dried under suitable conditions. Preferably, the concentration of dissolved or suspended solids in the feed stock is at least about 1 g/L, at least about 2 g/L, at least about 5 g/L, at least about 10 g/L, at least about 15 g/L, at least about 20 g/L, at least about 30 g/L, at least about 40 g/L, at least about 50 g/L, at least about 60 g/L, at least about 70 g/L, at least about 80 g/L, at least about 90 g/L, or at least about 100 g/L. The feed stock can be provided by preparing a single solution or suspension by dissolving or suspending suitable components (e.g. salts, excipients, other active ingredients) in a suitable solvent. The solvent, emulsion or suspension can be prepared using any suitable methods, such as bulk mixing of dry and/or liquid components or static mixing of liquid components to form a combination. For example, a hydrophilic component (e.g., an aqueous solution) and a hydrophobic component (e.g., an organic solution) can be combined using a static mixer to form a combination. The combination can then be atomized to produce droplets, which are dried to form respirable dry particles. Preferably, the atomization step is carried out immediately after combining the components in the static mixer. Alternatively, the atomization step is carried out in a bulk mixed solution.

[0105] The feedstock or components of the feedstock can be prepared using any suitable solvent, such as an organic solvent, an aqueous solvent or a mixture thereof. Suitable organic solvents that may be employed include, among others, alcohols, such as, for example, ethanol, methanol, propanol, isopropanol, butanol and the like. Other organic solvents include, but are not limited to, perfluorocarbons, dichloromethane, chloroform, ether, ethyl acetate, methyl tert-butyl ether and others. Co-solvents that may be employed include an aqueous and an organic solvent, such as, among others, the organic solvents described above. Aqueous solvents include water and buffered solutions.

[0106] The feed stock or components of the feed stock may have any desired pH, viscosity, or other property. If desired, a pH buffer may be added to the solvent or co-solvent or mixture formed. Generally, the pH of the mixture ranges from about 3 to about 8.

[0107] Dry particles and respirable dry powders can be manufactured and then separated, for example, by filtration or centrifugation through a cyclone, to provide a particle sample with a pre-selected size distribution. For example, greater than about 30%, greater than about 40%, greater than about 50%, greater than about 60%, greater than about 70%, greater than about 80% , or greater than about 90% of the dry respirable particles in a sample may have a diameter within the selected range. The selected range within which a given percentage of the dry respirable particles falls may be, for example, any of the size ranges described herein, such as a VMGD between about 0.1 to about 3 microns.

[0108] The invention also relates to respirable dry powders or respirable dry particles manufactured by preparing a feed stock solution, emulsion or suspension and spray drying the feed stock in accordance with the methods described herein. The feed stock may be prepared using (a) sodium chloride in an amount of about 79.5% to about 80.5% by weight (e.g., total solutes used to prepare the feed stock) and (b) leucine in an amount of at least about 19.5% to about 20.5% by weight (e.g., of the total solutes used to prepare the feed stock) and tiotropium bromide in an amount of about 0.01 % to about 0.5% by weight (e.g., total solutes used to prepare feed stock). All weight percentages are presented on a dry (anhydrous) basis.

[0109] In one embodiment, the respirable dry powders or respirable dry particles of the invention can be obtained by (1) preparing a feed stock comprising (a) a dry solute containing a weight percentage of the total dry solute of about 79 .5% to about 80.5% sodium chloride, about 19.5% to about 20.5% leucine, and from about 0.01% to about 0.5% tiotropium bromide , and (b) one or more solvents suitable for dissolving the solute and forming a feed stock, and (2) spray drying the feed stock. Various methods (e.g., static mixing, bulk mixing) can be used for mixing solutes and solvents to prepare feed stocks, which are known in the art. If desired, other suitable mixing methods can be used. For example, additional components that cause or facilitate mixing may be included in the feed stock. For example, carbon dioxide produces bubbles or effervescence and therefore can serve to promote physical mixing of the solute and solvent.

[0110] The diameter of dry respirable particles, for example, their VMGD, can be measured using an electrical zone detection instrument such as a Multisizer IIe (Coulter Electronic, Luton, Beds, England), or a laser diffraction instrument such as a HELOS system (Sympatec, Princeton, NJ) or a Mastersizer system (Malvern, Worcestershire, UK). Other instruments for measuring the geometric diameter of particles are well known in the art. The diameter of dry respirable particles in a sample will vary depending on factors such as particle composition and synthesis methods. The size distribution of respirable dry particles in a sample can be selected to allow for optimal deposition within target locations within the respiratory system.

[0111] Experimentally, the aerodynamic diameter can be determined through time-of-flight (TOF) measurements. For example, an instrument such as the Aerosol Particle Size Spectrometer (APS) (TSI Inc., Shoreview, MN) can be used to measure the aerodynamic diameter. APS measures the time required for individual dry respirable particles to pass between two fixed laser beams.

[0112] The aerodynamic diameter can also be experimentally determined using directly a conventional gravitational sedimentation method, in which the time required for a sample of dry respirable particles to settle at a certain distance is measured. Indirect methods for measuring mass mean aerodynamic diameter include the Andersen Cascade Impactor (ACI), next generation impactors (NGI), and multistage liquid collision (MSLI) methods. Methods and instruments for measuring the aerodynamic diameter of particles are well known in the art.

[0113] Packed density is a measure of the mass density of the envelope characterizing a particle. Packed density is accepted in area as an approximation of the mass density of a particle's envelope. The envelope mass density of a particle of a statistically isotropic form is defined as the mass of a particle divided by the minimum volume of the envelope sphere into which it can be introduced. Features that may contribute to low compacted density include irregular surface texture, high particle cohesiveness, and porous structure. Compacted density can be measured by means of instruments known to those skilled in the art such as a Dual Platform Microprocessor Controlled Tap Density Tester (Vankel, NC), a GeoPycTM instrument (Micrometrics Instrument Corp., Norcross, GA), or a SOTAX Tap Density Tester model TD2 (SOTAX Corp., Horsham, PA). Packed density can be determined using the USP Bulk Density and Packed Density Method, United States Pharmacopeia Convention, Rockville, MD, 10th Supplement, 4950-4951, 1999.

[0114] The fine particle fraction can be used as a way to characterize the aerosol performance of a dispersed powder. The fine particle fraction describes the size distribution of airborne dry respirable particles. Gravimetric analysis, using the cascade impactor, is a method of measuring the size distribution or fine particle fraction of airborne respirable dry particles. The Andersen cascade impactor (ACI) is an eight-stage impactor that can separate aerosols into nine distinct fractions based on aerodynamic size. The size cutoffs for each phase are dependent on the flow rate at which the ACI is operated. The ACI is composed of multiple stages consisting of a series of nozzles (i.e., a jet plate) and an impaction surface (i.e., an impaction disk). In each phase, an aerosol stream passes through the nozzles and impacts the surface. Dry respirable particles in the aerosol stream with a large and sufficient inertia will impact the plate. Smaller respirable dry particles that do not have sufficient inertia to impact the plate will remain in the aerosol stream and will be carried to the next phase. Each successive phase of the ACI has a high aerosol velocity at the nozzles so that smaller dry respirable particles can be collected in each successive phase. Specifically, the eight-stage ACI is calibrated so that the fraction of dust that is collected in stage 2 and all lower stages, including final collection, is composed of dry respirable particles that have an aerodynamic diameter less than 4.4 microns. The air flow of such a calibration is approximately 60 L/min.

[0115] If desired, a two-phase collapsed ACI can be used to measure the fine particle fraction. The two-phase collapsed ACI consists of only phases 0 and 2 of an eight-phase ACI, as well as the final collection filter, and allows the collection of two separate dust fractions. Specifically, a two-phase collapsed ACI is calibrated so that the fraction of dust that is collected in phase two is composed of dry respirable particles that have an aerodynamic diameter of less than 5.6 microns and greater than 3.4 microns. The dust fraction passing through phase two and depositing on the final collection filter is then composed of dry respirable particles with an aerodynamic diameter of less than 3.4 microns. The air flow of such a calibration is approximately 60 L/min.

[0116] FPF (<5.6) has been demonstrated to correlate the fraction of dust that is capable of acting on patients' lungs, while FPF (<3.4) has been demonstrated to correlate the fraction of dust that reaches the a patient's lung. These correlations provide a quantitative indicator that can be used in particle optimization.

[0117] The emitted dose may be determined using a method of USP section 601 Aerosols, Metered Dose Inhalers, and Dry Powder Inhalers, Delivered Dose Uniformity, Delivered Dose Sampling of Dry Powder Inhalers, United States Pharmacopoeia Convention, Rockville, MD, 13th Revision, 222-225, 2007. This method uses an in vitro device configured to mimic patient dosing.

[0118] An ACI can be used to approximate the emitted dose, which is here called recovered gravimetric dose and recovered analytical dose. “Recovered gravimetric dose” is defined as the ratio of the heavy dust in the filters of all stages of the ACI to the nominal dose. “Analytical dose recovered” is defined as the ratio of the powder recovered from the rinsing of all phases, the filters of all phases and induction from an ACI port to the nominal dose.

[0119] Another way of approximating the emitted dose is to determine how much the powder leaves its container, for example, capsule or blister, after firing a dry powder inhaler (DPI). This takes into account the percentage leaving the capsule, but does not take into account any dust deposited in the DPI. The mass of powder emitted is the difference between the weight of the capsule with the dose before firing the inhaler and the weight of the capsule after firing the inhaler. This measurement may be called capsule emitted powder mass (CEPM) or sometimes called “shot-weight”.

[0120] A multiphase liquid impactor (MSLI) is another device that can be used to measure the fine particle fraction. The multi-phase liquid impactor operates on the same principles as the ACI, although instead of eight phases, the MSLI has five. Additionally, each MSLI phase consists of an ethanol-moistened glass frit rather than a solid plate. The moistened phase is used to prevent particle bouncing and entrainment, which can occur when using ACI.

[0121] The new generation impactor (NGI) is a particle classifier cascade impactor for testing metered dose, dry powder and similar inhalation device.

[0122] Geometric particle size distribution can be measured for respirable dry powder after it is emitted from a dry powder inhaler (DPI) by use of a laser diffraction instrument such as the Malvern Spraytec. With the inhaler installed in the open bench configuration, an airtight seal is made on the air inlet side of the DPI causing the aerosol output to pass perpendicularly through the laser beam as an outward flow. In this way, known flow rates can be blown through the DPI by positive pressure to empty the DPI. The resulting geometric particle size distribution of the aerosol is measured by photodetectors with samples typically taken at 1000 Hz for the inhalation duration and the Dv50, GSD, FPF<5.0 μm measured and averaged over the inhalation duration.

[0123] The water content of dry respirable dust or dry respirable particles can be measured by a Karl Fisher titration machine, or by thermogravimetric analysis or thermal gravimetric analysis (TGA). The Karl Fischer titration uses colorimetric or volumetric titration to determine traces of water in a sample. TGA is a method of thermal analysis in which changes in the weight of materials are measured as a function of temperature (with constant heating rate), or as a function of time (with constant temperature and/or constant mass loss). TGA can be used to determine the water content or residual solvent content of the material being tested.

[0124] The invention also relates to a dry respirable powder or dry respirable particles produced using any of the methods described herein.

[0125] The respirable dry particles of the invention can also be characterized by the chemical, physical, aerosol components, and the solid state stability of the therapeutic agents and excipients that the respirable dry particles comprise. The chemical stability of constituent salts can affect important characteristics of respirable particles including shelf life, suitable storage conditions and environments for administration, biological compatibility, and the effectiveness of the salts. Chemical stability can be assessed using techniques well known in the art. An example of a technique that can be used to assess chemical stability is reversed-phase high-performance liquid chromatography (RP-HPLC).

Therapeutic Uses and Methods

[0126] The respirable dry powders and respirable dry particles of the invention are suitable for administration to the respiratory tract. The dry powders and dry particles of the invention may be administered to a subject in need thereof for the treatment of respiratory (e.g., lung) diseases, such as chronic bronchitis, emphysema, chronic obstructive pulmonary disease, asthma, hyperreactivity of the airways, seasonal allergic reaction, bronchiectasis, cystic fibrosis, parenchymal lung inflammatory conditions and the like, and for the treatment, reduction in the incidence or severity, and/or prevention of acute exacerbations of these chronic diseases, such as exacerbations caused by viral infections ( e.g., influenza virus, parainfluenza virus, respiratory syncytial virus, rhinovirus, adenovirus, metapneumovirus, coxsackie virus, echo virus, coronavirus, herpes virus, cytomegalovirus, and the like), bacterial infections (e.g., Streptococcus pneumoniae, which is commonly called pneumococcus, Staphylococcus aureus, Burkholderis ssp., Streptococcus agalactiae, Haemophilus influenzae, Haemophilus parainfluenzae, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Moraxella catarrhalis, Chlamydophila pneumoniae, Mycoplasma pneumoniae, Legionella pneumophila, Serratia marcescens, Mycobacterium tuberculosis, Bordetella pertussis, and the like), fungal infections (e.g., Histoplasma capsulatum, Cryptococcus neoformans, Pneumocystis jiroveci, Coccidioides immitis, and the like) or parasitic infections (e.g., Toxoplasma gondii, Strongyloides stercoralis, and the like), or environmental allergens and irritants (e.g. example, aeroallergens, including pollen and cat dander, airborne particulates, and the like). In a preferred embodiment, the lung disease is chronic bronchitis, emphysema, or chronic obstructive pulmonary disease. If desired, the respirable dry powders and respirable dry particles may be administered orally.

[0127] In another preferred embodiment, the lung disease is asthma.

[0128] In some aspects, the invention is a method for treating lung diseases, such as chronic bronchitis, emphysema, chronic obstructive pulmonary disease, asthma, airway hyperreactivity, seasonal allergic reaction, bronchiectasis, cystic fibrosis and similar, which comprise administering to the respiratory tract an effective amount of dry respirable particles or dry powder, to a subject in need thereof, as described herein. In a preferred embodiment, the lung disease is chronic bronchitis, emphysema, or chronic obstructive pulmonary disease.

[0129] In another preferred embodiment, the lung disease is asthma.

[0130] In other aspects, the invention is a method for treating, reducing the incidence or severity, or preventing acute exacerbations of a chronic lung disease, such as chronic bronchitis, emphysema, chronic obstructive pulmonary disease, asthma , airway hyperreactivity, seasonal allergic reaction, bronchiectasis, cystic fibrosis and the like, which comprise administering to the respiratory tract of a subject in need thereof an effective quantity of dry respirable particles or dry powder, as described herein . In a preferred embodiment, the lung disease is chronic bronchitis, emphysema, or chronic obstructive pulmonary disease.

[0131] In another preferred embodiment, the lung disease is asthma.

[0132] In other aspects, the invention is a method for reducing inflammation, including administering to the respiratory tract of a subject in need thereof an effective amount of dry particles or respirable dry powders, as described herein. Thus, dry particles and respirable dry powders can be used to broadly prevent or treat acute and/or chronic inflammations and, in particular, inflammations that characterize a number of lung diseases and conditions including, chronic bronchitis, emphysema, chronic obstructive pulmonary disease ( COPD), asthma, airway hyperreactivity, seasonal allergic reaction, bronchiectasis, cystic fibrosis (CF), parenchymal lung inflammatory diseases/conditions, and the like. In a preferred embodiment, the lung disease is chronic bronchitis, emphysema, or chronic obstructive pulmonary disease. Dry particles and dry powders can be administered to prevent the inflammation inherent in diseases such as COPD, asthma, and CF, and the increased inflammation caused by acute exacerbations of the diseases, both of which play a major role in the pathogenesis of these diseases.

[0133] In a preferred embodiment, the lung disease is chronic bronchitis, emphysema, or chronic obstructive pulmonary disease. In another preferred embodiment, the lung disease is asthma.

[0134] In other aspects, the invention is a method for alleviating the symptoms of a respiratory disease and/or a chronic lung disease, such as chronic bronchitis, emphysema, chronic obstructive pulmonary disease, asthma, airway hyperreactivity, seasonal allergic reaction, bronchiectasis, cystic fibrosis and the like, including administering to the respiratory tract of a subject in need thereof an effective amount of dry particles or respirable dry powders as described herein. In a preferred embodiment, the lung disease is chronic bronchitis, emphysema, or chronic obstructive pulmonary disease. In another preferred embodiment, the lung disease is asthma.

[0135] In other aspects, the invention is a method for improving the lung function of a patient with a respiratory disease and/or a chronic lung disease, such as chronic bronchitis, emphysema, chronic obstructive pulmonary disease, asthma, hyperreactivity respiratory tract, seasonal allergic reaction, bronchiectasis, cystic fibrosis and the like, including administering to the respiratory tract of a subject in need thereof an effective amount of dry particles or respirable dry powders, as described herein. Without wishing to be bound by theory, improving a patient's lung function over a period of time is a long-term way to treat a respiratory disease and prevent acute exacerbations. In a preferred embodiment, the lung disease is chronic bronchitis, emphysema, or chronic obstructive pulmonary disease. In another preferred embodiment, the lung disease is asthma.

[0136] Dry respirable particles and dry powders may be administered to the respiratory tract of a subject in need thereof using any suitable method, such as instillation techniques, and/or an inhalation device, such as a powder inhaler. dry (DPI) or metered dose inhaler (MDI). A variety of DPIs are available, such as the inhaler disclosed in US Patent Nos. 4,995,385 and 4,069,819, Spinhaler® (Fisons, Loughborough, United Kingdom), Rotahalers®, Diskhaler® and Diskus® (GlaxoSmithKline, Research Triangle Technology Park, North Carolina), FlowCaps® (Hovione, Loures, Portugal), Inhalators® (Boehringer-Ingelheim, Germany), Aerolizer® (Novartis, Switzerland), RS-01 high resistance and low resistance (Plastiape, Italy) and others known to those skilled in the art.

[0137] The following scientific journal articles are incorporated by reference for the complete overview of the following dry powder inhaler (DPI) configurations: 1) Single-dose Capsule DPI, 2) Multi-dose Blister DPI, and 3) Reservoir DPI multidoseDPI. N. Islam, E. Gladki, “Dry powder inhalers (DPIs)—A review of device reliability and innovation”, International Journal of Pharmaceuticals, 360(2008):1-11. H. Chystyn, “Diskus Review”, International Journal of Clinical Practice, June 2007, 61, 6, 1022-1036. H. Steckel, B. Muller, “In vitro evaluation of dry powder inhalers I: drug deposition of commonly used devices”, International Journal of Pharmaceuticals, 154(1997):19-29. Some representative capsule-based DPI units are RS-01 (Plastiape, Italy), Turbospin® (PH&T, Italy), Brezhaler® (Novartis, Switzerland), Aerolizer (Novartis, Switzerland), Podhaler® (Novartis, Switzerland) , HandiHaler® (Boehringer Ingelheim, Germany), AIR® (Civitas, Massachusetts), Dose One® (Dose One, Maine) and Eclipse® (Rhone Poulenc Rorer). Some representative unit dose DPIs are Conix® (3M, Minnesota), Cricket® (Mannkind, California), Dreamboat® (Mannkind, California), Occoris® (Team Consulting, Cambridge, United Kingdom), Solis® (Sandoz), Trivair® (Trimel Biopharma, Canada), Twincaps® (Hovione, Loures, Portugal). Some representative blister-based DPI units are Diskus® (GlaxoSmithKline (GSK), UK), Diskhaler® (GSK), Taper Dry® (3M, Minnisota), Gemini® (GSK), Twincer® (University of Groningen, Netherlands), Aspirair® (Vectura, UK), Acu-Breathe® (Respirics, Minnisota, USA), Exubra® (Novartis, Switzerland), Gyrohaler® (Vectura, UK), Omnihaler® (Vectura, UK), Microdose® (MicroDose Therapeutix, USA), Multihaler® (Cipla, India) Prohaler® (Aptar), Technohaler® (Vectura, UK), and Xcelovair® (Mylan, Pennsylvania). Some representative reservoir-based DPI units are Clickhaler® (Vectura), Next DPI® (Chiesi), Easyhaler® (Orion), Novolizer® (Meda), Pulmojet® (sanofi-aventis), Pulvinal® (Chiesi), Skyehaler ® (Skyepharma), Duohaler® (Vectura), Taifun® (Akela), Flexhaler® (AstraZeneca, Sweden), Turbuhaler® (AstraZeneca, Sweden), and Twisthaler® (Merck), and others known to those skilled in the art.

[0138] Generally, inhalation devices (e.g., DPIs) are capable of delivering a maximum amount of dry powder or dry particles in a single inhalation, which is related to the capacity of the blisters, capsules (e.g., size 000, 00, 0E, 0, 1, 2, 3, and 4, with respective volumetric capacities of 1.37ml, 950μl, 770μl, 680μl, 480μl, 360μl, 270μl, and 200μl) or other media containing dry particles or dry powder inside the inhaler. Preferably, the blister has a volume of about 360 microliters or less, about 270 microliters or less, or, more preferably, about 200 microliters or less, about 150 microliters or less, or about 100 microliters or less. Preferably, the capsule is a size 2 capsule, or a size 4 capsule. More preferably, the capsule is a size 3 capsule. Thus, delivery of a desired dose or effective amount may require two or more inhalations. Preferably, each dose that is administered to a subject in need thereof contains an effective amount of dry respirable particles or dry powders and is administered using no more than about 4 inhalations. For example, each dose of dry particles or dry respirable powders can be administered in a single inhalation or 2, 3, or 4 inhalations. Dry particles and respirable dry powders are preferably administered in a single step of active inspiration using a passive DPI. When this type of device is used, the subject's inhalation energy disperses dry respirable particles and sucks them into the respiratory tract.

[0139] The dry particles or respirable dry powders may be preferably released by inhalation to a desired area within the respiratory tract, as desired. It is well known that particles with an aerodynamic diameter (MMAD) of about 1 micron to about 3 microns can be released into the deep lung. Larger MMAD, for example, about 3 microns to about 5 microns can be released into the central and upper airways. Therefore, without wishing to be limited by theory, the invention has an MMAD of about 1 micron to about 5 microns, and preferably, about 2.5 microns to about 4.5 microns, which preferentially deposits more of the therapeutic dose into the central airways than in the upper airways or deep lung.

[0140] For dry powder inhalers, deposition in the oral cavity is dominated by inertial impact and thus characterized by the aerosol Stokes number (DeHaan et al. Journal of Aerosol Science, 35 (3), 309-331, 2003). For equivalent inhaler geometry, breathing pattern and oral cavity geometry, the Stokes number, and thus deposition in the oral cavity, is essentially affected by the aerodynamic size of the inhaled powder. Thus, factors contributing to the oral deposition of a powder include the size distribution of individual particles and the dispersibility of the powder. If the MMAD of the individual particles is very large, for example about 5 μm, then an increased percentage of dust will be deposited in the oral cavity. Likewise, if a powder has poor dispersibility, it is an indication that the particles will leave the powder inhaler dry and enter the oral cavity as clumps. The agglomerated dust will act aerodynamically as an individual particle as large as the agglomerate, therefore, even if the individual particles are small (e.g., MMAD of 5 microns or less), the size distribution of the inhaled dust may have an MMAD greater than 5 μm, leading to optimized deposition in the oral cavity.

[0141] Therefore, it is desirable to have a powder in which the particles are small, dense, and dispersible so that the powders consistently deposit in the desired region of the respiratory tract. For example, respirable dry powders containing respirable dry particles have an MMAD of about 5 microns or less, between about 1 micron and about 5 microns, preferably between about 2.5 microns and about 4.5 microns; are dense particles, e.g., have a high packed density and/or the envelope density is as desired, such as about 0.45 g/cm3 or more, about 0.45 g/cm3 to about 1.2 g/cm3 cm3, about 0.5 g/cm3 or more, about 0.55 g/cm3 or more, about 0.55 g/cm3 to about 1.0 g/cm3, or about 0.6 g/cm3 cm3 at about 1.0 g/cm3; and are highly dispersible (e.g., 1/4 bar or, alternatively, 0.5/4 bar of less than about 2.0, and preferably about 1.5 or less, or about 1.4 or less ). The compacted density and/or envelope density and MMAD are theoretically related to VMGD through the following formula:MMAD = VMGD*sqrt (envelope density or compacted density). If it is desired to deliver a high mass of therapeutic using a fixed volume dosing container, then particles of higher packed density and/or envelope density are desired.

[0142] Respirable dry powders containing respirable dry particles may also have a compact density of at least about 0.4 g/cm3, a compact density greater than 0.4 g/cm3, or a compact density greater than 0.4 g/cm3 to about 1.2 g/cm3.

[0143] The respirable dry powders and respirable dry particles of the invention can be used in compositions suitable for releasing drugs via the respiratory system. For example, such compositions may include mixtures of dry respirable particles of the invention and one or more other dry particles or powders, such as dry particles or powders that contain another active agent, or that consist of or consist essentially of one or more pharmaceutically acceptable excipients. . Respirable dry powders may include mixtures of dry particles with lactose, such as large lactose-carrying particles that are larger than 10 microns, 20 microns to 500 microns, and preferably between 25 microns and 250 microns.

[0144] Respirable dry powders and respirable dry particles suitable for use in the methods of the invention can travel through the upper airways (i.e., the oropharynx and larynx), the lower airways, which include the trachea followed by bifurcations in the bronchi and bronchioles, and through the terminal bronchioles, which in turn divide into respiratory bronchioles leading to the final respiratory zone, the alveoli, or the deep lung. In one embodiment of the invention, the majority of the mass of dry powders or respirable particles is deposited in the deep lung. In another embodiment of the invention, the release is mainly in the central airways. In another modality, the release is to the upper respiratory tract. In a preferred embodiment, the majority of the mass of respirable dry powders or particles is deposited in the conducting airways.

[0145] Appropriate intervals between doses that provide the desired therapeutic effect can be determined based on the severity of the condition, the subject's general well-being and the subject's tolerance to dry particles and respirable dry powders and other considerations. Based on these and other considerations, a doctor can determine appropriate intervals between doses. Generally, respirable dry particles and respirable dry powders are administered once, twice or three times a day as needed.

[0146] If desired or indicated, the dry respirable particles and dry respirable powders described herein can be administered with one or more other therapeutic agents. The other therapeutic agents may be administered by any suitable route, such as orally, parenterally (e.g., intravenous, intra-arterial, intramuscular, or subcutaneous injection), topically, by inhalation (e.g., intrabronchial, intranasal or oral inhalation, intranasal drops ), rectally, vaginally and the like. The dry respirable particles and dry powders can be administered prior to, substantially simultaneously with, or subsequent to the administration of the other therapeutic agent. Preferably, the dry respirable particles and dry powders and the other therapeutic agent are administered so as to provide substantial overlap in their pharmacological activities.

EXEMPLIFICATION

[0147] The materials used in the following Examples and their sources are listed below. Sodium chloride, and L-leucine were obtained from Sigma-Aldrich (St. Louis, MO), Spectrum Chemicals (Gardena, CA), or Merck (Darmstadt, Germany). Tiotropium bromide was obtained from RIA International (East Hanover, NJ). Ultrapure water (ASTM Type II) was obtained from a water purification system (Millipore Corp., Billerica, MA), or equivalent.

Methods:

[0148] Diameter or Geometric Volume. Volume mean diameter (x50 or DV50), which can also be referred to as the volume mean geometric diameter (VMGD), was determined using a laser diffraction technique. The equipment consisted of a HELOS diffractometer and a RODOS dry powder disperser (Sympatec, Inc., Princeton, NJ). The RODOS disperser applies a shear force to a particle sample, controlled by the pressure regulator (typically set at 1.0 bar, with maximum orifice ring pressure) of the incoming compressed dry air. Pressure settings can be varied to vary the amount of energy used to disperse the powder. For example, the dispersion energy can be modulated by changing the pressure regulator from 0.2 bar to 4.0 bar. The powder sample is dispensed from a micro-spatula into the RODOS funnel. The scattered particles travel through a laser beam, where the resulting diffracted light pattern produced is collected, typically using an R1 lens, by an array of detectors. The ensemble diffraction pattern is then translated into a volume-based particle size distribution using the Fraunhofer diffraction model, on the basis that smaller particles diffract light at larger angles. Using this method, the geometric standard deviation (GSD) for the volume diameter was also determined.

[0149] The volume mean diameter can also be measured using a method in which powder is emitted from a dry powder inhalation device. The equipment consisted of a Spraytec (Malvern, Worcestershire, UK) particle size laser diffraction system, “Spraytec”. Powder formulations were manually placed into size 3 HPMC capsules (Capsugel V-Caps) with the weight of the contents measured gravimetrically using an analytical balance (Mettler Tolerdo XS205). A capsule-based passive dry powder inhaler (RS01 Model 7, high resistance, Plastiape S.p.A.) was used and had a specific resistance of 0.036 kPa^LPM-1. Flow rate and inhaled volume were set using a time-controlled solenoid valve with flow control valve (TPK2000, Copley Scientific). The capsules were placed in the dry powder inhaler, punctured, and the inhaler sealed inside a cylinder. The cylinder was connected to a positive pressure air source with a constant air flow through the system measured with a mass flow meter and its duration controlled with a time-controlled solenoid valve. The output of the dry powder inhaler was exposed to ambient pressure and the resulting aerosol jet passed through the laser diffraction particle sizer (Spraytec) in its open bench configuration, before being captured by a vacuum extractor. A constant airflow rate through the system was initiated using the solenoid valve and particle size distribution was measured using the Spraytec at 1 kHz for the duration of a single inhalation maneuver, with a minimum of 2 seconds. Calculated particle size distribution parameters included volume mean diameter (Dv50) and geometric standard deviation (GSD) and fine particle fraction (FPF) of particles less than 5 micrometers in diameter. Upon completion of the inhalation duration, the dry powder inhaler was opened, the capsule removed and reweighed to calculate the mass of powder that had been emitted from the capsule during the inhalation duration (mass of powder emitted from the capsule or CEPM ).

[0150] Fine particle fraction. The aerodynamic properties of powders dispersed from an inhalation device were evaluated with an Andersen Mk-II 1 ACFM Cascade Impactor (Copley Scientific Limited, Nottingham, United Kingdom) (ACI) or a New Generation Impactor (Copley Scientific Limited , Nottingham, United Kingdom) (NGI). The ACI instrument was run under controlled environmental conditions of 18 to 25°C and relative humidity (RH) between 25 and 35%. The instrument consists of eight phases that separate aerosol particles based on inertial impact. In each phase, the aerosol flow passes through a set of nozzles and impacts on a corresponding impaction plate. Particles that have sufficiently small inertia will continue with the aerosol stream to the next phase, while the remaining particles will impact the plate. In each successive phase, the aerosol passes through the nozzles at a higher speed and aerodynamically smaller particles will be collected on the plate. After the aerosol passes through the final phase, a filter collects the smallest particles that remain, in the so-called “final collector filter”. Gravimetric and/or chemical analyzes can then be performed to determine particle size distribution. A short stack cascade impactor, also referred to as a collapsed cascade impactor, is also utilized to allow for reduced work time to evaluate the two aerodynamic particle size cutoffs. With this collapsed cascade impactor, phases are eliminated except those required to establish fine and coarse particle fractions.

[0151] The impaction techniques used allow the collection of two or eight separate powder fractions. Capsules (HPMC, Size 3; Capsugel Vcaps, Peapack, NJ) were manually filled with powder to a specific weight and placed in a portable, breath-activated dry powder inhalation (DPI) device, the heavy-duty RS01 DPI (Plastiape , Osnago, Italy). The capsule was punctured and the powder was sucked through the cascade impactor operated at a flow rate of 60.0 L/min for 2.0 s. At this flow rate, the calibrated cutoff diameters for the eight phases are 8.6, 6.5, 4.4, 3.3, 2.0, 1.1, 0.5 and 0.3 microns and for the two phases used with the short stack cascade impactor, based on the Andersen Cascade impactor, the cutoff diameters are 5.6 microns and 3.4 microns. The fractions were collected by placing the filters in the device and determining the amount of dust impacting them by gravimetric measurements or chemical measurements on an HPLC. The fine particle fraction of the total powder dose (FPFTD) less than or equal to an effective aerodynamic cutoff diameter was calculated by dividing the mass of powder recovered from the desired phases of the impactor by the total mass of particles in the capsule. Results are presented for the eight normal phases of the stack cascade impactor as the fine particle fraction of less than 4.4 microns (FPFTD <4.4 microns) and the fine particle fraction of less than 2.0 microns ( FPFTD <2.0 microns), and the two-phase short stack cascade impactor as the fine particle fraction of less than 5.6 microns (FPFTD <5.6 microns) and the fine particle fraction of less than 3 .4 microns (FPFTD <3.4 microns). The fine particle fraction can alternatively be calculated in relation to the emitted or recovered dose of powder by dividing the mass of powder recovered from the desired phases of the impactor by the total mass of powder recovered in the impactor.

[0152] Likewise, for FPF measurements using the NGI, the NGI instrument was run under controlled environmental conditions of 18 to 25°C and relative humidity (RH) between 25 and 35%. The instrument consists of seven stages that separate aerosol particles based on inertial impact and can be operated with a variety of air flow rates. At each stage, the aerosol stream passes through a set of nozzles and collides with a corresponding impaction surface. Particles having sufficiently small inertia will continue with the aerosol stream to the next phase, while remaining particles will impact the surface. In each successive phase, the aerosol passes through the nozzles at a higher speed and aerodynamically smaller particles are collected on the plate. After the aerosol passes through the final phase, a micro-orifice collector collects the remaining smaller particles. Chemical analyzes can then be performed to determine particle size distribution. Capsules (HPMC, Size 3; Capsugel Vcaps, Peapack, NJ) were manually filled with powder of a specific weight and placed in a portable, breath-activated, RS01 DPI heavy-duty dry powder inhalation (DPI) device (Plastiape, Osnago, Italy). The capsule was punctured and the dust was aspirated through the cascade impactor operated at a specific flow rate of 2.0 liters of inhaled air. At the specified flow rate, the cutoff diameters for the phases were calculated. The fractions were collected by placing moistened filters in the device and determining the amount of dust that impinges on them through chemical measurements on an HPLC. The fine particle fraction of the total powder dose (FPFTD) less than or equal to an effective cutoff aerodynamic diameter was calculated by dividing the mass of powder recovered from the desired phases of the impactor by the total mass of particles in the capsule. Results for NGI were reported as the fraction of fine particles less than 5.0 microns (FPFTD <5.0 microns)

[0153] Aerodynamic diameter. The mass mean aerodynamic diameter (MMAD) was determined using information obtained from the Andersen cascade impactor (ACI). The accumulated mass below the cutoff diameter is calculated for each phase and normalized by the recovered powder dose. The MMAD of the powder is then calculated by linear interpolation of the phase cutoff diameters that fall within the 50th percentile. An alternative method of measuring MMAD is with the Next Generation Pharmaceutical Impactor (NGI). Like ACI, MMAD is calculated as the accumulated mass below the cutoff diameter is calculated for each phase and normalized by the recovered powder dose. The MMAD of the powder is then calculated by linear interpolation of the phase cutoff diameters that fall within the 50th percentile.

[0154] Fine particle dose. The fine particle dose FPD(FPD) is determined using the information obtained by the ACI. Alternatively, the FPD is determined using the information obtained by the NGI. The fine particle dose indicates the mass of one or more therapeutic agents in a specific size range and can be used to predict the mass that will reach a certain region of the respiratory tract. The dose of fine particles can be measured gravimetrically or chemically. If measured gravimetrically, since it is assumed that the dry particles are homogeneous, the mass of powder in each phase and collection filter can be multiplied by the fraction of therapeutic agent in the formulation, to determine the mass of this therapeutic. If measured chemically, the powder from each phase or filter is collected, separated, and analyzed, for example, in an HPLC to determine the amount of therapeutic agent. The accumulated mass deposited on the final collection filter, and phases 6, 5, 4, 3, and 2 for a single dose of powder, contained in one or more capsules, activated in the ACI is equal to the dose of fine particles lower than 4.4 microns (FPD <4.4 microns). The accumulated mass deposited on the final collection filter, and phases 6, 5 and 4, for a single dose of powder, contained in one or more capsules, activated in the ACI is the same for the dose of fine particles less than 2.0 microns (FPD <2.0 microns). The quotient of these two values is expressed as FPD <2.0 μm/FPD <4.4 μm. The higher the ratio, the greater the percentage of therapeutics entering the lungs, which are expected to penetrate the alveolar regions of the lungs. The lower the ratio, the lower the percentage of therapeutics entering the lungs that are expected to penetrate the alveolar regions of the lungs. For some therapies that target the central or conducting airways, lower proportions such as less than 40%, less than 30%, or less than 20% are desirable. For other therapies targeting the deep lung, higher proportions such as 40% or more, 50% or more, or 60% or more are desirable. Similarly, for FPD measurements using the NGI, the NGI instrument was run as described in the fine particle fraction description in the Exemplification section. The accumulated mass deposited on each of the phases at the specified flow rate is calculated and the accumulated mass corresponding to a particle diameter of 5.0 micrometers is interpolated. This accumulated mass for a single dose of powder, contained in one or more capsules, driven into the NGI is equal to the dose of fine particles less than 5.0 microns (FPD<5.0 microns).

[0155] Geometric Diameter or Issued Volume. The volume mean diameter (Dv50) of the powder after being emitted from a dry powder inhaler, which can also be referred to as the volume mean geometric diameter (VMGD), was determined using a laser diffraction technique through the Spraytec diffractometer (Malvern, Inc.). The Powder was filled into size 3 capsules (V-Caps, Capsugel) and placed in a capsule-based dry powder inhaler (RS01 Model 7 high resistance, Plastiape, Italy), or DPI, and the DPI sealed inside a cylinder . The cylinder was connected to a positive pressure air source with a constant air flow through the system measured with a mass flow meter and its duration controlled with a time-controlled solenoid valve. The outlet of the dry powder inhaler was exposed to ambient pressure and the resulting aerosol jet passed through the diffraction particle sizer (Spraytec) laser in its open bench configuration, before being captured by a vacuum extractor. Constant air flow through the system was initiated using the solenoid valve. A constant air flow rate was drawn through the DPI typically at 60L/min for a defined duration, typically 2 seconds. Alternatively, the air flow rate sucked through the DPI was sometimes run at 15L/min, 20L/min, or 30L/min. The resulting geometric particle size distribution of the aerosol was calculated from the software based on the scattering pattern measured at photodetectors with samples typically taken at 1000Hz for the duration of inhalation. The measured Dv50, GSD, FPF <5.0μm were then averaged over the inhalation duration.

[0156] The Emitted Dose (ED) refers to the mass of therapeutic agent that exits a suitable inhaler device after a trigger or dispersion event. ED is determined using a method based on USP Section 601 Aerosols, Metered-Dose Inhalers and Dry Powder Inhalers, Delivered-Dose Uniformity, Sampling the Delivered Dose from Dry Powder Inhalers, United States Pharmacopeia convention, Rockville, MD, 13th Revision, 222 -225, 2007. The contents of the capsules are dispersed using the RS01 HR inhaler at a pressure drop of 4kPa and a typical flow rate of 60 LPM and the emitted powder is collected on a filter in a filter holder sampling device. The sampling apparatus is washed with a suitable solvent such as water and analyzed using an HPLC method. For gravimetric analysis a shorter length filter holder sampling apparatus is used to reduce deposition on the apparatus and the filter is weighed before and after to determine the mass of dust released from the DPI onto the filter. The emitted dose of the therapeutic agents is then calculated based on the therapeutic content of the released powder. The emitted dose can be reported as the mass of therapeutic agent released from the DPI or as a percentage of the filled dose.

[0157] Mass of powder emitted in a capsule. A measure of the emission properties of the powders was determined using information obtained from tests on the Andersen Cascade Impactor or geometric diameter emitted by the Spraytec. The weight of the filled capsule was recorded at the start of the run and the final weight of the capsule was recorded at the conclusion of the run. The difference in weight represented the amount of powder emitted from the capsule (CEPM or mass of powder emitted in capsule). CEPM was reported as a mass of dust or as a percentage by dividing the amount of dust emitted from the capsule by the mass of the total initial particle in the capsule. Although the standard CEPM was measured at 60L/min, it was also measured at 15L/min, 20L/min, or 30L/min.

[0158] Compacted density. Compacted density was measured using a modified method requiring smaller amounts of powder, following USP <616> with the replacement of a 1.5cc microcentrifuge tube (Eppendorf AG, Hamburg, Germany) or a 0.3cc section of a Disposable polystyrene serological micropipette (Grenier Bio-One, Monroe, NC) with polyethylene caps (Kimble Chase, Vineland, NJ) to cap both ends and retain powder. Instruments for measuring compacted density known to those skilled in the art include, among others, a Dual Platform Microprocessor Controlled Tap Density Tester (Vankel, Cary, NC) or a SOTAX Tap Density Tester model TD2 (Horsham, PA). Packed density is an approximate standard measure of the mass density of the envelope. The mass density of the envelope of an isotropic particle is defined as the mass of the particle divided by the minimum spherical envelope volume within which it can be enclosed.

[0159] Apparent density. Bulk density was estimated prior to the compacted density measurement procedure by dividing the weight of the powder by the volume of the powder, as estimated using volumetric measuring equipment.

[0160] Thermogravimetric Analysis. Thermogravimetric analysis (TGA) was performed using a Q500 Thermogravimetric Analyzer (TA Instruments, New Castle, DE). The samples were placed in an open aluminum DSC container, with the tare weight previously recorded by the instrument. The following method was employed: Ramp of 10.00°C/min from room temperature (~35°C) to 200°C. Weight loss has been reported as a function of temperature up to 150°C. TGA allows the calculation of the water content of the dry powder.

[0161] Tiotropium content using HPLC. Tiotropium content was measured using a high performance liquid chromatography (HPLC) system with an ultraviolet (UV) detector. The HPLC method was performed using an HPLC system with UV detection (HPLC-UV; Waters, Milford, MA) with a Waters Xterra MS C18 column (5μm, 3x100mm; Waters, Milford, MA) to identify and quantify tiotropium in a range from 0.03 μg/mL to 1.27 μg/mL. The HPLC-UV system was configured with 100 μL injection volume, 40°C column temperature, 240 nm detection wavelength, and isocratic elution with a 0.1% trifluoroacetic acid mobile phase (Fisher Scientific, Pittsburgh, PA) and acetonitrile (Fisher Scientific, Pittsburgh, PA) (85:15) to determine tiotropium content over a 10-minute run time. The results are reported as tiotropium content and tiotropium bromide content.

[0162] Preparation of liquid raw material for spray drying. Particle spray drying requires that the ingredients of interest be solubilized in solution or suspended in a uniform and stable suspension. Sodium chloride, leucine and tiotropium bromide are sufficiently soluble in water to prepare solutions suitable for spray drying. Alternatively, ethanol or another organic solvent can be used.

[0163] Spray drying using Niro spray dryer. The dried powders were produced by spray drying using a Niro Mobile Minor spray dryer (GEA Process Engineering Inc., Columbia, MD) with dust collection from a cyclone, a product filter, or both. Atomization of the liquid feed was accomplished using a two-fluid co-current nozzle from Niro (GEA Process Engineering Inc., Columbia, MD) or a two-fluid nozzle Spraying System (Carol Stream, IL) with gas cap. 67147 and 2850SS fluid cap, although other two-fluid nozzle configurations are also possible. In some embodiments, the two-fluid nozzle may be in an internal mixing configuration or an external mixing configuration. Other atomization techniques include rotary atomization or a pressure nozzle. The liquid feed was inserted using gear pumps (Cole-Parmer Instrument Company, Vernon Hills, IL) directly into the two-fluid nozzle or into a static mixer (Charles Ross & Son Company, Hauppauge, NY) immediately prior to introduction into the nozzle. of two fluids. An additional liquid feeding technique includes feeding from a pressurized container. Nitrogen or air may be used as a drying gas, provided that the moisture in the air is at least partially removed before use. Nitrogen or pressurized air can be used as the atomizing gas feed for the two-fluid nozzle. The process gas inlet temperature can range from 70°C to 300°C and the outlet temperature from 30°C to 120°C with a liquid feedstock rate of 10 mL/min to 100 mL/min. Gas delivery from the two-fluid atomizer can vary depending on nozzle selection and for the Niro two-fluid co-current nozzle can vary between 5 kg/h to 50 kg/h or for the two-fluid nozzle Spraying Systems with gas cap 67147 and 2850SS fluid cap can range from 30 g/min to 150 g/min. The atomization gas rate can be adjusted to achieve a certain gas to liquid mass ratio, which directly affects the size of the droplet created. The pressure inside the drying drum can vary from +3 “WC to -6 “WC. The pulverized dry powders can be collected in a container at the outlet of the cyclone, in a cartridge or bag filter, or from both a cyclone or cartridge or bag filter.

[0164] Process gas as used in these descriptions refers to drying gas. The two-fluid Spray Systems nozzles were a series of 1/4J nozzles.

[0165] Spray drying using Büchi Spray Dryer. Dry powders were prepared by spray drying in a Büchi B-290 Mini Spray Dryer (BÜCHI Labortechnik AG, Flawil, Switzerland), with dust collection from a standard or high-performance cyclone. The system was run with air or nitrogen as well as with drying gas and atomization in open cycle mode (single pass). When performed using air, the system used the Büchi B-296 dehumidifier to ensure stable temperature and humidity of the air used for spray drying. Additionally, when the relative humidity in the room exceeded 30% RH, an LG outdoor dehumidifier (model 49007903, LG Electronics, Englewood Cliffs, NJ) was running constantly. When run using nitrogen, a pressurized source of nitrogen was used. Additionally, the system vacuum was adjusted to maintain system pressure at -2.0 water columns. Atomization of the fed liquid used a Büchi two-fluid nozzle with a diameter of 1.5 mm or a Schlick 970-0 atomizer with a 0.5 mm insertion liquid (Düsen-Schlick GmbH, Coburg, Germany). Process gas inlet temperature can vary from 100°C to 220°C and outlet temperature from 30°C to 120°C with a liquid raw material flow rate of 3mL/min to 10mL/min. The two-fluid atomizing gas ranged from 25mm to 45mm (300LPH to 530LPH) for the Büchi two-fluid nozzle and for the Schlick atomizer an atomizing air pressure of up to 0.3 bar. The vacuum cleaner rate ranged from 50% to 100%.

[0166] Spray drying using ProCepT Formatrix. The dried powders were prepared by spray drying in a ProCepT Formatrix R&D spray dryer (ProCepT nv, Zelzate, Belgium). The system was run in open circuit configuration using ambient air in a manufacturing location controlled to <60% RH. The drying gas flow rate can vary from 0.2 to 0.5 m3/min. The bi-fluid nozzle was equipped for atomization with 0.15-1.2 mm liquid tips. The atomizing gas pressure can vary from about 0.5 bar to 6 bar. The system was equipped with small or medium cyclone. The inlet temperature of the spray dryer can range from about 100°C to 190°C, with an outlet temperature of about 40°C to about 95°C. The flow rate of liquid raw material can vary from about 0.1 to 15mL/min. Process parameters were controlled via the ProCepT human-machine interface (HMI) and all parameters were recorded electronically.

[0167] Clinical measurements. Clinical measurements reported below include the maximum concentration of tiotropium in the bloodstream (Cmax), the area under the curve for tiotropium in the bloodstream (AUC), and a patient's forced expiratory volume in one second (FEV1). Cmax is the maximum concentration of a therapeutic agent, such as tiotropium, in the bloodstream. It provides pharmacokinetic (PK) information. It is measured in picograms per milliliter (pg/mL). AUC indicates an individual's total systemic exposure to a therapeutic agent, such as tiotropium, over the indicated period of time. Here, AUC was measured during the following time periods, 0-2 hours, 0-6 hours, and 0-24 hours. AUC is measured in picogram-hours per milliliter (pg*h/mL). FEV1 is a person's forced expiratory volume in one second, and can be used to determine the effect of administering a therapeutic agent, such as tiotropium, to a patient. It provides pharmacodynamic (PD) information. FEV1 is measured in liters (L).

Example 1. Tiotropium and dry powder formulations containing salt. A. Powder preparation.

[0168] The raw material solutions were prepared and used to manufacture dry powders consisting of pure, dry particles containing tiotropium bromide, sodium chloride and leucine. Powders were prepared in triplicate. Table 2 lists the components of the raw material formulations used in the preparations of dry powders consisting of dry particles. Weight percentages are presented on a dry basis. Table 2. Raw material compositions

[0169] The raw material solutions that were used for spray-dried particles were prepared as follows. For Formulation I, the liquid raw material was batch mixed, the total solid concentration was 30g/L, the amount of tiotropium bromide in the solution was 0.012g/L, the amount of sodium chloride in the solution was 23.990g/L, the amount of leucine in the solution was 5.998g/L, and the final aqueous raw material was clear. For formulation V, the batch liquid raw material was mixed, the total solid concentration was 30g/L, the amount of tiotropium bromide in the solution was 0.066g/L, the amount of sodium chloride in the solution was of 23.947g/L, the amount of leucine in the solution was 5.989g/L, and the final raw material was clear. Raw material volumes ranged from 0.720 to 1.800L, which supported manufacturing campaigns of 2 to 5 hours.

[0170] The dry powders of Formulations I and V were manufactured from these raw materials obtained by spray drying in the Büchi B-290 Mini Spray Dryer (BÜCHI Labortechnik AG, Flawil, Switzerland), with powder collection in a cyclone. The system was executed in the open cycle module (single pass), using nitrogen as drying and atomization gas. Atomization of the liquid feed used a Schlick 970-0 atomizer with 0.5 mm of liquid inserted. The system vacuum was adjusted to maintain the system pressure at -2.0’’ water column.

[0171] The following spray drying conditions were followed to manufacture the dry powders. For formulations I and V, the liquid raw material solids concentration was 30g/L, the gas inlet temperature in the process was 178°C to 182°C, the gas outlet temperature in the process was 77°C, the drying gas flow rate was 18.0kg/h, the atomization gas flow rate was 1.824kg/h, the atomization gas return pressure at the atomizer inlet was 34 psig to 37 psig and the flow rate of liquid raw material was 6.0mL/min. The resulting dry powder formulations are reported in Table 3. Table 3. Dry powder compositions, on a dry basis

B. Powder characterization.

[0172] Size and density characteristics are found in Table 4. The VMGD of Formulations I and V using the RODOS at a dispersion energy of 1 bar were both 2.30 micrometers, with a standard deviation of 0.04 and 0.03, respectively, indicating that the process was reproducible with triplicate productions from spray drying. The range at 1 bar of 1.45 and 1.47 for Formulations I and V, respectively, indicates a relatively narrow size distribution. The 1 bar/4 bar dispersibility ratio of 1.21 and 1.22 for Formulations I and V, respectively, and the 0.5 bar/4 bar dispersibility ratio of 1.29 for both formulations indicates that they are relatively independent of dispersion energy, a desirable feature that allows relatively similar therapeutic dose to be administered to a varied population of patients.

[0173] The geometric particle size and mass of powder emitted by the capsule (CEPM) measured and/or calculated at simulated patient flow rates of 60 liters per minute (LPM) and 20 LPM were evaluated for the two formulations and reported in the Table 4. Formulations I and V had a CEPM of 100% at 60 LPM and 97% at 20 LPM. Formulation I had a Dv50 of 2.43 microns at 60 LPM and 2.63 microns at 20 LPM, and Formulation V had a Dv50 of 2.39 microns at 60 LPM and 2.62 microns at 20 LPM. The small change in CEPM and geometric size from 60 LPM to 20 LPM indicates that the dry powder formulations are relatively independent of the patient's inspiratory flow rate, indicating that patients breathing at different flow rates can receive a relatively similar therapeutic dose.

[0174] The aerodynamic particle size, fine particle fractions and fine particle doses measured and/or calculated with a two-stage and/or eight-stage Anderson Cascade Impactor (ACI -2 and ACI -8), are reported in Table 4. The fine particle fraction of the total dose less than 2.0, 3.4, 4.4, and 5.6 microns, for Formulation I were 14%, 44%, 56.7% , and 72%, respectively, and for Formulation V they were 14%, 45%, 58.8%, and 73%, respectively. It should be noted that ACI-2 was run with just one capsule containing about 10 mg of dry powder, while ACI-8 was run with 2 capsules, for a total of 20 mg of dry powder. The fine particle dose less than 4.4 micrometers (FPD < 4.4) for Formulation I and V were 3.77 micrograms tiotropium and 21.65 micrograms tiotropium released from two 10 mg capsules of powder each or 1.89 micrograms and 10.82 micrograms per capsule respectively.

[0175] The fine particle dosage for Formulation I and V both indicates a high percentage of the nominal dose that is filled into the capsule that reaches the impactor phases (57% and 59%) and therefore would be expected to be released to the lungs. This reflects a significant improvement in the effectiveness of the formulations of the invention over the commercially available product SPIRIVA (tiotropium bromide) lactose blend-based dry powder HANDIHALER (dry powder inhaler). The SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) product has a nominal dose of 18 micrograms tiotropium inserted into each capsule dose, but delivers a fine particle dose (FPD < 4.4) between 2.5 and 3.7 micrograms from each filled capsule, as calculated by Shur et al., AAPS Journal, 2012 and Chodosh et al., Journal of Aerosol Medicine 2001.

[0176] The MMAD of Formulation I and V were 3.20 microns and 3.17 microns, respectively, indicating deposition in the central and conducting airways. The “FPD <2.0 micron/FPD <4.4 micron” ratio for Formulation I and V were 25% and 24%, respectively, indicating that 75% and 76%, respectively, of FPD less than 4.4 microns have an FPD of 2.0 microns or greater, another indication that the majority of dry powder should be deposited in the central and conducting airways.

[0177] Properties related to the apparent and compacted densities of the formulations are reported in Table 4. The apparent density of both formulations was 0.26g/cm3 and the compacted density of Formulation I was 0.68g/cm3 and of Formulation V was 0.69g/cm3. For Formulations I and V, the Hausner ratios were 2.58 and 2.70, respectively, and the Carr ratio was 61.22 and 62.57, respectively.

[0178] The water content of Formulations I and V were measured with TGA and are reported in Table 4. They are 0.07% and 0.08%, respectively.

[0179] The tiotropium bromide contents of Formulations I and V were measured by HPLC-UV and are reported in Table 4. They are 0.040% and 0.221%, respectively. Table 4. Size and density characteristics of Formulations I and V

Example 2. Formulation V reduces specific airway resistance after methacholine challenge in healthy mice.

[0180] In order to determine the effectiveness of tiotropium bromide present in Formulation V, lung function tests were performed on healthy mice one hour after treatment with Formulation V. Treatments were performed in a full-body exposure chamber using a capsule-based dry powder inhaler system. The dose was varied by changing the number of capsules used for each exposure, in this case, 2 or 3 90mg capsules for an estimated expected dose of 4.4μg or 6.6μg of tiotropium bromide, respectively. A lung function test was performed by measuring specific airway resistance (sRaw) in mice via dual-chamber plethysmography. Baseline sRaw measurements were taken for 5 minutes, followed by 5 minutes of measurement after nebulization of 0mg/ml and 100mg/ml methacholine chloride (MCh) dissolved in 0.9% sodium chloride in the main chamber.

[0181] The results are presented in Table 5. Treatment with both 4.4μg and 6.6μg of tiotropium bromide resulted in a greater than 40% reduction in sRaw compared to untreated mice, p=0.035 and p=0.032 , respectively, after challenge with 100mg/ml MCh. There were no significant changes in sRaw at baseline or after challenge with 0mg/ml MCh. It is known from the literature, for example, “Effect of tiotropium bromide on airway inflammation and remodeling in a mouse model of asthma”, Clinical and Experimental Allergy (40: 1266-1275), that tiotropium bromide results in improved lung function during MCh challenge by antagonizing the M3 muscarinic receptor, the same receptor that recognizes MCh, ultimately leading to bronchoconstriction. Furthermore, tiotropium bromide has been shown to improve lung function, resulting in lower sRaw values, for animals and humans challenged with inhaled MCh in 0.9% sodium chloride. The results reported in Table 5, specifically unchanged sRaw values at baseline and at 0mg/ml together with a significant reduction in sRaw after challenge with 100mg/mL MCh. Formulation V reduced sRaw following MCh challenge in healthy mice. Table 5. MCh challenge following Formulation V treatment in healthy mice. Data are represented as Mean ± SD; *p<0.05 Example 3. Formulation of dry powders containing Tiotropium in combination with other pharmacologically active agents.

A.Powder preparation.

[0182] The raw material solutions were prepared in order to manufacture dry powders consisting of dry particles containing a sodium salt, a non-salt excipient, tiotropium and optionally additional active pharmaceutical agents. Table 6 lists the components of the raw material formulations used in the preparation of dry powders consisting of dry particles. Weight percentages are given on a dry basis. Table 6. Raw Material Compositions of sodium salt with tiotropium and in combination with other pharmaceutically active agents.

[0183] The raw material solutions were prepared according to the parameters in Tables 7 and 8.

[0184] Table 7. Formulation Conditions Table 8. Formulation Conditions

[0185] For all formulations, the liquid raw material was mixed in batches

[0186] Formulations VI to VIII of dry powders were produced by spray drying in the Mini Spray Dryer Büchi B-290 (BÜCHI Labortechnik AG, Flawil, Switzerland), with powder collection from a high-performance cyclone in a container of 60mL glass. The system utilized the Büchi B-296 dehumidifier and an LG outdoor dehumidifier (model 49007903, LG Electronics, Englewood Cliffs, NJ) was run constantly. Atomization of the liquid feed used a Büchi two-fluid nozzle with a diameter of 1.5 mm. The two-fluid atomization gas was set at 40mm (667 LPH). The aspiration rate was adjusted to 80% (32m3/h) for Formulation VI; 70% for Formulations VII and VIII. Air was used as drying gas and atomizing gas. Table 9 below includes details on spray drying conditions. Table 9. Spray Drying Process Conditions

B. Powder characterization.

[0187] Physical and aerosol properties of the powder are summarized in Tables 10 to 14 below. Values with ± indicate standard deviation of the reported value. Two-phase ACI-2 results are reported in Table 10 for FPFTD < 3.4μm and FPFTD < 5.6μm. Formulations VI through VIII had an FPFTD < 3.4 μm greater than 20% and an FPFTD < 3.4 μm greater than 30%. Formulation VI had an FPFTD < 3.4 μm greater than 45%. Formulations VI through VIII had an FPFTD < 5.6 μm greater than 40% and an FPFTD < 5.6 μm greater than 60%. Table 10. Aerodynamic Properties

[0188] Formulations VI through VIII have a compacted density greater than 0.35g/cm3, and Formulations VI and VII have a compacted density greater than 0.40g/cm3. Formulations VI and VII have a compacted density greater than 0.50g/cm3. Formulations VI to VIII have a Hausner ratio greater than or equal to 1.5. Formulations VII and VIII have a Hausner ratio greater than 2.0. Formulation VII has a Hausner ratio of 3.07 (see Table 11). Table 11. Density Properties

[0189] Table 12 shows that Formulations VI through VIII had Dv50 of less than 2.0 microns at 60 LPM. Formulations VI through VIII had a Dv50 of less than 6.0μm at 15 LPM. Formulation VII had a Dv50 of less than 5.0 μm at 15 LPM. Table 12. Geometric Diameters

[0190] Table 13 shows that Formulations VI through VIII have a capsule emitted particle mass (CEPM) greater than 96% at 60 LPM. Formulations VI through VIII have a CEPM greater than 90% at 15 LPM. Table 13. Dispersibility Properties

[0191] Table 14 shows that Formulations VI to VIII have a Dv50 of less than 2.0 μm when using the RODOS at a setting of 1.0 bar. Formulations VI through VIII have a RODOS to 0.5bar/4bar ratio of less than 1.4, and Formulations VI and VIII have a RODOS to 0.5bar/4bar ratio of less than 1.3. Formulations VI through VIII have a RODOS to 1bar/4bar ratio of less than or equal to about 1.1. Table 14. Dispersibility Properties (Geometric Diameter using SQUEEGES) Example 4. Effect of a dry powder based on monovalent cations of tiotropium bromide (Formulation VI) on airway hyperreactivity in a mouse model of ovalbumin allergic asthma.

[0192] In a mouse model of ovalbumin allergic asthma, protocols for sensitization and subsequent challenge with OVA, and lung function tests are described in Examples 6 to 8 of PCT Publication WO 2012/044736 “Monovalent Cation Dry Powders” and are incorporated herein for reference in its entirety.

[0193] It was known from the literature that tiotropium bromide (TioB) improves lung function, resulting in lower sRaw values, for animals and humans challenged with methacholine chloride (MCh) at 0.9% of sodium chloride for inhalation. (Ohta, S. et al. (2010), “Effect of tiotropium bromide on airway inflammation and remodeling in a mouse model of asthma”, Clinical and Experimental Allergy. 40: 1266-1275).

[0194] While the effects of TioB on sRaw were known from the literature, the effect of co-formulating the TioB formulation with a sodium salt was unknown. Formulation VI (34.47% leucine, 65.42% NaCl, and 0.113% tiotropium bromide, w/w on a dry basis) was tested and compared to Placebo-B dry powder (98% leucine, 2 % NaCl, w/w on dry basis). The results for the pulmonary function tests are presented in Table 15. Table 15. Effect of Formulation VI on Specific Airway Resistance.

[0195] These data show that Formulation VI significantly reduced sRaw during MCh challenge compared to Placebo-B (p < 0.00001). Example 5. Pharmacokinetic (PK) and pharmacodynamic (PD) effects of dry powders based on monovalent tiotropium bromide cations (Formulations I-IV).

[0196] In a pilot study, four doses (two per cohort) of a dry powder based on the monovalent cation of tiotropium bromide, Formulations I-IV, were evaluated. Plasma levels of tiotropium bromide were measured over time (Figure 1) and the bronchodilatory effect was measured (Figure 2). The subjects were patients with moderate to severe COPD. Single doses and matching placebos of Formulations I-IV were administered in a double-blind inter-vivo crossover design. The study was designed to enroll a total of 24 COPD subjects randomized to 2 cohorts of 12, each taking part in a 2-way dosing regimen. Within each cohort up to 10 subjects were to receive the active agent (Formulations I-IV, comprising tiotropium bromide) and 2 were to receive the placebo. Formulation I provides a nominal dose of 3μg, Formulation II provides a nominal dose of 6μg, Formulation III provides a nominal dose of 9μg, and Formulation IV provides a nominal dose of 12μg, as indicated in Table 16.

[0197] The data used in this example, including the tables and figures, represents unaudited data from a clinical trial. The formulations used in this example were manufactured with a ProCepT Formatrix R&D spray dryer. Table 16. Formulations I-IV and Placebo.

[0198] Cohort 1 received Formulation I or placebo during the first treatment period (12 subjects: 2 placebo, 10 Formulation I), followed by a treatment-free period of at least seven days. After a safety review, Cohort 2 received Formulation II or placebo during the first treatment period (12 subjects: 2 placebo, 10 Formulation II), followed by a treatment-free period of at least seven days. After a safety review and treatment-free period, Cohort 1 returned to receive Formulation III or placebo for the second treatment period (11 subjects: 2 placebo, 9 Formulation III). After a final safety review and treatment-free period, Cohort 2 returned to IV Formulation or placebo for the second treatment period (10 subjects: 2 placebo, 8 IV Formulation).

[0199] Serum levels of tiotropium bromide were determined using LC-MS/MS (liquid chromatography tandem mass spectrometry) methods well known in the art, for example, as described in Nilsson et al. PLoS One, 5 (7): e11411 (2010). The following analytes were detected: Table 17. Tiotropium analytes.

[0200] Pulmonary function tests were performed with a standard calibrated spirometer. As a measure of the bronchodilatory effects of Formulations I-IV in human subjects, FEV1 (forced expiratory volume in one second) was used. The predicted FEV1 was obtained using the normal prediction equations by the ERS (European Respiratory Society) adjusting for race, sex and age. Quanjer et al. “Lung volume and forced ventilatory flows.” Eur Respir J. 6: Suppl. 16, 5-40 (1993). Spirometry measurements were conducted in accordance with 2005 ATS/ERS (American Thoracic Society/European Respiratory Society) guidelines. The single highest FEV1 value and the single highest FVC (forced vital capacity) value of acceptable and reproducible maneuvers were reported and the FEV1/FVC ratio was determined.

[0201] Unaudited data obtained from the pilot study suggests that tiotropium bromide, exemplified by Formulations I-IV, was effectively delivered to the lungs in monovalent cation-based dry powders, and tiotropium bromide can be measured in the blood. Figure 1A illustrates the geometric mean PK profile of each nominal dose level (Formulations I-IV, 3μg, 6μg, 9μg, and 12μg, respectively) over a 6-hour period. The maximum concentration for tiotropium (Cmax) was detected at approximately 5 minutes after administration. This profile is consistent with the PK profile of the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Figure 1B illustrates the average area under the curve (AUC0-2h) for each Formulation, I-IV. For comparison, the known profile of single doses of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) is indicated by a gray boxed area. Data for the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) can be obtained from the FDA website: www.accessdata.fda.gov/scripts/cder/drugsatfda/. The AUC0-2h for Formulations I-IV supported the profile of the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler).

[0202] Figure 2 illustrates pharmacodynamic data suggesting that all Formulations I-IV improved lung function, each exerting a measurable bronchodilatory effect as measured by increased peak FEV1 over the first 6h after dosing and increased trough of FEV1 24h after dosing. Formulation I (a nominal dose of 3μg tiotropium) resulted in a measurable and sustained increase in FEV1. Formulations II-IV (nominal dose of 6, 9, and 12μg tiotropium, respectively) resulted in greater improvements in FEV1, compared to Formulation I. The profiles obtained for Formulations II-IV were quite similar. The single-dose improvement in lung function of Formulations I-IV is comparable to the single-dose SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) improvement in lung function, as described, for example, by Maesen et al, “Tiotropium bromide, a new long-acting antimuscarinic bronchodilator: a pharmacodynamics study in patients with chronic obstructive pulmonary disease (COPD)” Eur. Respir. J, 8:1506-13 (1995). However, improvements can be achieved with significantly lower nominal doses when compared to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Example 6. An in vitro test comparing the aerodynamic particle sizes of Formulation II tiotropium bromide to the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler)

[0203] The in vitro mass mean aerodynamic diameter (MMAD) test, also referred to as the aerodynamic particle size distribution (aPSD) test, was performed compared to Formulation II with SPIRIVA (tiotropium bromide) HANDIHALER (powder inhaler dry). The objectives of this study were: i) to inform the development of a formulation that achieved similar fine particle dose delivery across a range of flow rates relevant to patients with COPD and ii) to determine whether Formulation II, representative of all Formulations I- IV, was less dependent on a patient's inspiratory flow rate than SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). This MMAD test was performed across a range of flow rates relevant to the intended COPD patient population.

[0204] Formulation II was produced and inserted into size 3 HPMC capsules for dispersion in the RS01 dry powder inhaler. SPIRIVA (tiotropium bromide) was purchased and dispersed from the HANDIHALER (dry powder inhaler). A New Generation Multi-Phase Impactor (NGI) was used to determine the mass distributed over various aerodynamic diameters, the fine particle fraction (FPF) less than 5 micrometers in diameter, and the fine particle dose FPD(FPD) less than 5 micrometers in diameter. Due to the difference in airflow resistance of the two dry powder inhalers, the formulations were compared at similar pressure drops across the inhaler rather than at the same airflow rate. Distributions are shown at a pressure drop of 4kPa across each dry powder inhaler (See Fig. 3 and Table 18 below). For Formulation II, using the RS01 HR device, this pressure drop correlated with an inspiratory flow rate of 60 LPM. For SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), this pressure drop correlated with an inspiratory flow rate of 39 LPM. The test was carried out with replicates, n=5 for Formulation II, using the RS01 HR inhaler and n=3 for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Similar aerodynamic particle size distributions are observed for the 2 products and the fine particle dose (FPD <5.0μm) was found to be comparable for Formulation II using the RS01 inhaler and the SPIRIVA (tiotropium bromide) HANDIHALER ( dry powder), although the nominal dose of Formulation II was 5.8 micrograms and the nominal dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) was 18 micrograms. This means that the delivery efficiency for Formulation II using the RS01 inhaler was over three times more efficient than the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). This result is further illustrated by the difference in the fraction of fine particles (FPF <5.0μm) relative to the nominal dose, which was 54.8% for Formulation II in the RS01 HR, and 15.0% for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). (See Table 19 below). Loss of drug product for the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) was attributed to the relatively larger amounts of therapeutic product left in the capsule, device, mouthpiece adapter, induction port, and in the preseparator. (See Fig. 4 and Table 20 below). A preseparator was not included in the NGI test with Formulation II with the RS01 inhaler because carrier particles were not present in the spray-dried formulation. Table 18. Mass distribution of tiotropium by NGI phase at 4 kPa, for Formulation II using the RS01 inhaler for a nominal dose of 5.8μg of tiotropium and for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) for nominal dose of 18 micrograms (μg). Table 19. Nominal dose of tiotropium and FPF less than 5.0 microns as a percentage of the nominal dose using the RS01 inhaler and the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Table 20. Mass distribution of tiotropium in the various components before entering the NGI at 4kPa for Formulation II using the RS01 inhaler and SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler).

[0205] The aerosol test was performed over a range of peak inspiratory flow (PIF) relevant to the intended population of patients with COPD. A flow rate range of 20L/min to 55L/min, with a midpoint of 39L/min was selected to test SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which covers the range of PIF that has been measured for patients COPD with the product and is specified in the product label in the United States for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). For Formulation II, the corresponding flow range was calculated from the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) by differential pressure combination based on the different resistances of the DPIs, resulting in a PIF range of 28L/min to 84L/min with a midpoint of 60 LPM. The DPI used for Formulation II was the RS01 HR inhaler, while the DPI used for the SPIRIVA (tiotropium bromide) product was the HANDIHALER (dry powder inhaler).

[0206] Formulation II was produced and inserted into size 3 HPMC capsules for dispersion in the RS01 dry powder inhaler. SPIRIVA (tiotropium bromide) was purchased and dispersed from the HANDIHALER (dry powder inhaler). The fine particle dose (FPD <5.0 microns) is presented at pressure drops of 1, 4, and 8 kPa across the dry powder inhaler for Formulation II and SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler ) as determined using an NGI. The fine particle dose was shown to be less sensitive to flow rate for Formulation II compared to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). The flow rate dependence of the fine particle dose (less than 5 micrometers) for the two products is shown in FIG. 5 and Table 21 (n = 3-5 replicates; values shown are mean ± standard deviation). Formulation II, using the RS01 HR inhaler, demonstrated to be less sensitive to the patient's simulated inspiratory flow rate, even with the low DPI pressure drop of 1 kPa. These results indicate that Formulation II using RS01 would provide improved nominal dose delivery efficiency compared to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) and more consistent pulmonary delivery across the patient population. including those with low PIF due to highly compromised lungs. Table 21. Flow rate dependence of Formulation II using RS01 versus SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) Note: N=3 replicates for both inhalers in all conditions, except N=5 for RS01 in the 4 kPa condition. Example 7. Pharmacokinetic and pharmacodynamic (PD) effects of dry powders based on monovalent tiotropium bromide cation (Formulations I - III).

[0207] In a Phase Ib study, three strong doses of a dry powder based on the monovalent cation of tiotropium bromide, Formulations I - III were evaluated. This study was designed to enroll up to 40 patients with COPD in a randomized, double-blind, single-dose 5-way crossover study. Formulations I-III, placebo, and the open-label comparison product were the 5 study arms. There was a free period of at least 7 days between doses. Serum levels of tiotropium bromide were measured over time (see Fig. 6 and Table 23) and the bronchodilatory effect was measured (Fig. 7). The subjects were patients with moderate to severe COPD. Formulation I provides a nominal dose of 3μg, Formulation II provides a nominal dose of 6μg, and Formulation III provides a nominal dose of 9μg, as indicated in Table 22. Table 22. Formulations I-III, Placebo and SPIRIVA (tiotropium bromide ) HANDIHALER (dry powder inhaler) Table 23. Serum PK values at Cmax and AUC values varying over the period Note: All time points and Cmax are in pg/mL and AUC is in pg*h/mL

[0208] Serum levels of tiotropium bromide were determined using LC-MS/MS (Liquid Chromatography-Tandem Mass Spectrometry) methods well known in the art, for example, as described in Nilsson et al. PLoS One, 5(7): e11411 (2010). The following analytes were detected (Table 24): Table 24. Tiotropium Analytes.

[0209] Pulmonary function tests were performed using a standard calibrated spirometer. As a measure of the bronchodilatory effects of Formulations I-III, in human subjects, FEV1 (forced expiratory volume in one second) was used. The predicted FEV1 was obtained using the normal prediction equations by the ERS (European Respiratory Society) adjusting race, sex and age. Quanjer et al. “Lung volume and forced ventilatory flows.” Eur Respir J. 6: Suppl. 16, 5-40 (1993). Spirometry measurements were conducted in accordance with 2005 ATS/ERS (American Thoracic Society/European Respiratory Society) guidelines. The single highest FEV1 value and the single highest FVC (forced vital capacity) value of acceptable and reproducible maneuvers were reported and the FEV1/FVC ratio determined. The data is reported in FIG. 7.

[0210] Unaudited data obtained from the Phase Ib study suggests that tiotropium bromide, exemplified by Formulations I-III, was effectively delivered to the lungs in monovalent cation-based dry powders, and tiotropium bromide can be measured in blood.

[0211] Fig. 6 illustrates the geometric mean PK profile of each nominal dose level (Formulations I - III, 3μg, 6μg, 9μg, respectively), and the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) throughout over a period of 6 hours. The maximum concentration for tiotropium (Cmax) was detected at approximately 5 minutes after administration. SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Serum levels for Formulations I to III increased with dose and exhibited similar kinetics to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Cmax and AUC0-2h are similar between Formulation I and SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). AUC0-24h are similar between Formulation II and SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler).

[0212] Fig. 7 illustrates pharmacodynamic data suggesting that all Formulations I-III improved lung function, each exerting a measurable bronchodilatory effect as measured by the increase in peak FEV1 over the first 6h post dose and increased trough of FEV1 24h after the dose. Formulations I-III resulted in a measurable and sustained increase in FEV1, with the data showing a significant and sustained increase in FEV1 compared to placebo. Formulation I combining the changes in FEV1 for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), in both “peak” and “valley” improvements. Formulations II and III resulted in greater improvement in lung function than SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler).

[0213] These clinical data demonstrate the ability to combine the pulmonary dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) with a significantly lower nominal dose. Formulation I is comparable to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) across multiple parameters and achieves similar improvement in lung function to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) at total systemic exposure significantly lower. Formulation II, which has a similar total systemic exposure to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), as seen by AUC0-24h results in better benefits in lung function at the same exposure dose.

A. AUDITED CLINICAL DATA

[0214] As mentioned above, in this example, the results presented were obtained from unaudited data from the Phase Ib study. Below is a presentation of audited data from the same Phase Ib study as presented previously in this example, which support the observations and conclusions written above. The above data represents unaudited clinical data. It should be noted that the data in Tables 23 and 25 and Fig. 6 are geometric mean values and the data in Table 26 and FIG.7 are mean values. It should be noted that the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) was the comparator for Formulations I - III in the clinical trial. SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) has a nominal dose of 18 micrograms of tiotropium, which is mixed with 5.5mg of lactose monohydrate, and is essentially a 5.5mg full weight capsule. Table 25. Serum PK values at Cmax and AUC values (values reported as geometric mean values. SD = Standard Deviation) Note: All time points and Cmax are in pg/mL and AUC is in pg*hr/mL

[0215] Table 25 reports the audited clinical data for serum PK values (Cmax and AUC) during different time points up to 24 hours after dosing. The Cmax value of Formulation I was 7.85pg/mL ± 6.84pg/mL and corresponds closely to that of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which was 7.22 pg/mL ± 6 .85 pg/ml. The Cmax values of Formulations II and III were 15.9 pg/ mL ± 13.4 pg/ ml and 29.6 pg/ mL ± 18.9 pg/ ml, respectively, which were both higher than the Cmax of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). The AUC0-24 of Formulation I was 8.77 ± 7.00 pg*h/mL, which was about 1/3 of the AUC0-24h of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which was 24.7 ± 12.8 pg*h/ mL. This means that the systemic exposure of Formulation I was about 1/3 that of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). The AUC0-24h of Formulation II was 21.2 ± 12.0 pg*h/mL, which was approximately the same AUC0-24h as SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which was 24. 7 ± 12.8 pg*h/mL. This means that the systemic exposure of Formulation II was approximately the same as that of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). The AUC0-24h of Formulation III was 38.0 ± 18.4 pg*h/mL, which was greater than the AUC0-24h of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). This means that the systemic exposure of Formulation III was greater than that of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler).

[0216] The audited clinical data presented in Table 25 confirms the conclusions made from the unaudited data presented in Table 23 and FIG. 6. Table 26. Absolute value in FEV1 and change in FEV1 from baseline at 6 hours and 24 hours after dosing. (Values reported as mean values. SD = Standard Deviation)

[0217] Table 26 reports the audited clinical data for FEV1 values during different time points up to 24 hours. At the 6 hour time point, administration of Formulation I caused a change in FEV1 of 0.239 L ± 0.163 L, which corresponds closely to the result observed for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), which caused a change in FEV1 of 0.240 L ± 0.189 L, indicating a similar improvement in lung function at 6 hours. Formulations II and III caused a change in FEV1 of 0.294 L ± 0.166 L and 0.292 L ± 0.168 L, respectively, both of which are greater than the change in FEV1 of 0.240 L ± 0.189 L observed for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), indicating a greater improvement in lung function within 6 hours. At the 24-hour time point, which was the point of observation of the FEV1 trough, administration of Formulation I caused a change in FEV1 of 0.151 L ± 0.120 L, which was slightly less than the result observed for SPIRIVA (bromide tiotropium) HANDIHALER (dry powder inhaler), which caused a change in FEV1 of 0.169 L ± 0.157 L, indicating that Formulation I caused an improvement in lung function at 24 hours, but it was slightly less than that observed for SPIRIVA ( tiotropium bromide) HANDIHALER (dry powder inhaler). Formulations II and III caused a change in FEV1 of 0.218 L ± 0.145 L and 0.198 L ± 0.164 L, respectively, both of which are greater than the change in FEV1 of 0.169 L ± 0.157 L observed for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), indicating greater improvement in lung function within 24 hours.

[0218] The audited clinical data presented in Table 26 confirms the conclusions made from the unaudited data presented in FIG. 7.

[0219] Both unaudited and audited data obtained from the Phase Ib study suggest that tiotropium bromide, exemplified by Formulations I-III, was effectively delivered to the lungs in monovalent cation-based dry powders, and that Tiotropium bromide can be measured in the blood. The audited clinical data corresponds closely to the unaudited clinical data and served to confirm the conclusions drawn from the unaudited data.

[0220] These clinical data demonstrate the ability to match the pulmonary dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) with a significantly lower nominal dose. Formulation I was comparable to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) in improving lung function over the 24-hour time period, SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) as demonstrated by similar improvements in FEV1 at the 6-hour time point and the 24-hour trough time point. This similar improvement in lung function was achieved with similar values for Cmax, yet with a significantly lower total systemic exposure, as seen by AUC0-24h results of about 1/3 of those observed for SPIRIVA (tiotropium bromide) HANDIHALER ( dry powder inhaler). Formulation II, which has a similar total systemic exposure to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler), as seen by the AUC0-24h results in increased improvement in lung function at the same exposure dose, for example, change in FEV1 at 6 hour time point and 24 hour trough time point. Example 8. An In Vitro Test Comparing the Aerodynamic Particle Sizes of Formulation II Tiotropium Bromide from RS01 UHR2 with SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler)

[0221] The above example was substantially repeated, as described in Example 6, with a different model of dry powder inhaler releasing Formulation II. In Example 6, Formulation II was delivered using the RS01 HR dry powder inhaler, while in the present example, Formulation II was delivered from the RS01 UHR2 dry powder inhaler, which has a greater resistance to airflow than the RS01 HR model. In vitro aerodynamic particle size distribution (aPSD) testing was performed which compared the released form of Formulation II from the RS01 UHR2 dry powder inhaler to that of the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). The objectives of this study were: i) to inform the development of a formulation that achieved similar fine particle dose delivery across a range of flow rates relevant to patients with COPD and ii) to determine whether Formulation II, representative of all Formulations I to IV , was less dependent on the patient's inspiratory flow rate than SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). This aPSD test was performed across a range of flow rates relevant to the intended COPD patient population.

[0222] Formulation II was produced and inserted into size 3 HPMC capsules for dispersion in the RS01 UHR2 dry powder inhaler. SPIRIVA (tiotropium bromide) was purchased and dispersed from the HANDIHALER (dry powder inhaler). A New Generation Multi-Phase Impactor (NGI) was used to determine the mass mean aerodynamic diameters, the fine particle fraction (FPF) less than 5 micrometers in diameter, and the fine particle dose FPD(FPD) less than 5 micrometers in diameter. As both RS01 UHR2 and SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) dry powder inhalers had similar airflow resistances, the formulations were compared at the same airflow rates and also with similar pressure drop across the inhaler. . Particle size distributions are presented at a pressure drop of 4 kPa across each dry powder inhaler, which corresponds to 39 LPM across each dry powder inhaler. (See Fig. 8 and Table 27 below). The test was carried out with replicates, n = 3 for Formulation II, using the RS01 UHR2 inhaler and n = 3 for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Similar aerodynamic particle size distributions are observed for the 3 products and the fine particle dose (FPD <5.0μm) was shown to be comparable for Formulation II using the RS01 inhaler and the SPIRIVA (tiotropium bromide) HANDIHALER ( dry powder), although the nominal dose of Formulation II was 5.8 micrograms and the nominal dose of SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) was 18 micrograms. This means that the delivery efficiency for Formulation II using the RS01 UHR2 inhaler was more than three times more efficient than SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). This result is further illustrated by the difference in the fine particle fraction (FPF <5.0μm) in relation to the nominal dose, which was 51.4% for Formulation II in the RS01 UHR2 and 15.0% for SPIRIVA (bromide tiotropium) HANDIHALER (dry powder inhaler). (See Table 28 below.) Drug product loss for the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) was attributed to the relatively higher values of therapeutics left in the capsule, device, mouthpiece adapter, induction port and pre-separator. (See FIG. 9 and Table 29 below). A preseparator was not included in NGI's testing of Formulation II with the RS01 UHR2 inhaler because carrier particles were not present in the spray-dried formulation. Table 27. Tiotropium mass distribution by NGI phase at 4 kPa for Formulation II using the RS01 UHR2 inhaler per nominal dose of tiotropium of 5.8 μg and the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) of 18μg nominal dose. Table 28. Tiotropium nominal dose and FPF less than 5.0 microns as a percentage of the nominal dose using the RS01 UHR2 inhaler and the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). Table 29. Mass distribution of Tiotropium in the various components before entry into the NGI at 4kPa for Formulation II using the RS01 UHR2 inhaler and the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler).

[0223] The aerosol test was performed over a range of peak inspiratory flows (PIF) relevant to the intended population of patients with COPD. A flow range of 20 L/min to 55 L/min, with a midpoint of 39 L/min (corresponding to 1, 4 and 8 kPa) was selected to test the SPIRIVA (tiotropium bromide) HANDIHALER (powder inhaler dry) and Formulation II from RS01 UHR2, which covers the PIF range that was measured for patients with COPD, with the product and is specified in the product leaflet for SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) in the United States.

[0224] Formulation II was produced and inserted into size 3 HPMC capsules for dispersion in the RS01 UHR2 dry powder inhaler. SPIRIVA (tiotropium bromide) was purchased and dispersed from the HANDIHALER (dry powder inhaler). The fine particle dose (FPD <5.0 microns) is presented at 1, 4, and 8 kPa pressure drops across the dry powder inhaler for Formulation II from both the RS01 and SPIRIVA (tiotropium bromide ) HANDIHALER (dry powder inhaler) as determined using an NGI. The fine particle dose was shown to be less flow sensitive for Formulation II in the RS01 UHR2 inhaler compared to the SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler). The dose-dependent flow rate of fine particles (less than 5 micrometers) for the two products is shown in FIG. 10 and Table 30 (n = 3 replicates; values shown are the mean ± standard deviation). Formulation II, using the RS01 UHR2 inhaler, proved to be less sensitive to the patient's simulated inspiratory flow, even with the low pressure drop at the DPI of 1 kPa. These results indicate that Formulation II, using the RS01 UHR2 inhaler would provide both improved efficiency in delivering the nominal dose, compared to SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler) and a more consistent lung delivery throughout the patient population, including those with low PIF due to high lung involvement. Table 30. Flow rate dependence of Formulation II using RS01 UHR2 versus SPIRIVA (tiotropium bromide) HANDIHALER (dry powder inhaler)

Claims (22)

1. Container characterized by the fact that it comprises a respirable dry powder consisting of homogeneous respirable dry particles comprising sodium chloride, leucine and tiotropium bromide, wherein the sodium chloride content is 75% to 82% by weight, leucine is 15% to 25% by weight, tiotropium bromide is 0.01% to 0.5% by weight, and optionally one or more additional therapeutic agents up to 20% by weight, wherein all percentages are percentages by weight on a dry basis and all components of the dry respirable particles add up to 100%, wherein the container contains a nominal dose of tiotropium between 1.5 and 12 micrograms, and wherein the container can be contained in a powder inhaler dry. 2. Container according to claim 1, characterized by the fact that the container is a capsule or blister, and wherein the container contains 15 mg of dry powder or less. 3. Container according to claim 1, characterized in that the container contains a nominal dose of tiotropium between 3 and 12 micrograms. 4. Container according to claim 1, characterized in that the container contains a nominal dose of tiotropium between 3 and 9 micrograms. 5. Container according to claim 1, characterized in that the container contains a nominal dose of tiotropium between 3 and 6 micrograms. 6. Container, according to claim 1, characterized by the fact that the dry breathable powder has a fine particle dose of less than 5.0 microns and between 1 microgram and 5 micrograms of tiotropium. 7. Container according to claim 1, characterized in that the respirable dry powder has a fine particle dose of less than 4.4 microns and between 1 microgram and 4 micrograms of tiotropium. 8. Container according to claim 1, characterized in that the respirable dry powder has a fine particle dose of less than 4.4 microns and between 1.5 micrograms and 3.5 micrograms of tiotropium. 9. Container according to claim 1, characterized in that it is a capsule and in which the capsule contains less than 6 mg of dry powder. 10. Container according to claim 1, characterized by the fact that the sodium chloride content is 79.5% to 80.5% by weight and the leucine content is 19.5% to 20.5% by weight. % by weight. 11. Container according to claim 1, characterized by the fact that the dry respirable particles have a volume median geometric diameter (VMGD) of 5 micrometers or less. 12. Container, according to claim 1, characterized by the fact that the respirable dry powder has a mass median aerodynamic diameter (MMAD) between 1 and 5 microns. 13. Container according to claim 1, characterized by the fact that the dry respirable particles have a dispersibility rate of 1/4 bar (1/4 bar) less than 1.5, measured by laser diffraction. 14. Container according to claim 1, characterized in that the dry respirable particles have a tap density of 0.6 g/cm3 to 1.2 g/cm3. 15. Container according to claim 1, characterized by the fact that the one or more additional therapeutic agent is one or more corticosteroids. 16. Container characterized by the fact that it comprises a respirable dry powder consisting of homogeneous respirable dry particles comprising sodium chloride, leucine and tiotropium bromide, wherein the sodium chloride content is 75% to 82% by weight, the leucine content is 15% to 25% by weight, the tiotropium bromide content is 0.01% to 0.5% by weight, and optionally one or more additional therapeutic agents up to 20% by weight, in that all percentages are percentages by weight on a dry basis and all components of the dry respirable particles add up to 100%, wherein the container contains a nominal dose of tiotropium between 0.5 and 6 micrograms, and wherein the container may be contained in a dry powder inhaler. 17. Container according to claim 16, characterized in that the container contains a nominal dose of tiotropium between 0.5 and 3 micrograms. 18. Container, according to claim 16, characterized by the fact that the dry breathable powder has a fine particle dose of less than 5.0 microns and between 1 microgram and 5 micrograms of tiotropium. 19. Container according to claim 16, characterized in that the respirable dry powder has a fine particle dose of less than 4.4 microns and between 1.5 micrograms and 3.5 micrograms of tiotropium. 20. Use of an effective amount of a dry respirable powder as defined in claim 1, characterized in that it is in the manufacture of a medicament for treating a respiratory disease selected from the group consisting of COPD, chronic bronchitis, emphysema , asthma, cystic fibrosis or non-cystic fibrosis bronchiectasis. 21. Use of an effective amount of a dry respirable powder as defined in claim 1, characterized by the fact that it is in the manufacture of a medicament for treating or reducing the incidence or severity of an acute exacerbation of a respiratory disease selected from the group consisting of COPD, chronic bronchitis, emphysema, asthma, cystic fibrosis, or non-cystic fibrosis bronchiectasis. 22. Use according to claim 20, characterized by the fact that the respiratory disease is COPD.