System for treating pulmonary infections

Abstract

To provide a system for treating pulmonary infection.SOLUTION: According to the present invention, there is provided a method for treating various pulmonary infections including mycobacterial infections by providing a system for delivery of an aerosolized liposomal formulation via inhalation. In one aspect, the present invention provides a system for treating or providing prophylaxis against a pulmonary infection. In one embodiment, the system comprises a pharmaceutical formulation comprising: a liposomal complexed aminoglycoside (here, the formulation is a dispersion (e.g., a liposomal solution or suspension), the lipid component of the liposome consists of electrically neutral lipids); and a nebulizer which generates an aerosol of the pharmaceutical formulation at a rate greater than about 0.53 g per minute.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of priority from U.S. Provisional Patent Application No. 61 / 649,830, filed on May 21, 2012, the entire content of which is incorporated herein by reference.

Background Art

[0002] Background of the Invention In certain techniques suitable for administration by inhalation, liposomes are used, and in the lungs, lipid complexes supply the therapeutic effect of the drug over an extended period of time. These techniques also make it possible to provide drugs with sustained activity, target drugs to the disease site, and enhance the uptake of drugs into the disease site.

[0003] The inhalation delivery of liposomes is complicated by the fact that liposomes are susceptible to shear - induced stress during atomization, which can cause changes in their physical characteristics (e.g., encapsulation efficiency, size). However, as long as the changes in characteristics are reproducible and meet the acceptance criteria, they do not necessarily impede pharmaceutical development.

[0004] In patients with cystic fibrosis (CF), thick mucus and / or sputum secretions in the lungs, frequent secondary infections, and biofilms formed by bacterial colonization are seen. All of these fluids and substances are barriers to the effective targeting of infections by aminoglycosides. Liposomal aminoglycoside formulations may be useful in combating bacterial biofilms.

Summary of the Invention

Means for Solving the Problems

[0005] Summary of the Invention The present invention provides a method for treating various pulmonary infections, including mycobacterial infections (e.g., pulmonary infections caused by nontuberculous mycobacteria, herein also referred to as nontuberculous mycobacteria (NTM) infections), by providing a system for delivering aerosolized liposome formulations by inhalation. For example, the systems and methods provided herein are for pulmonary nontuberculous mycobacterial infections, such as pulmonary M. avium, M. avium subsp. hominissuis (MAH), M. abscessus, M. chelonae, M. bolletii, M. kansasii, M. ulcerans, M. avium, M. avium complex (MAC) (M. avium and M. intracellulare), M. conspicuum, M. kansasii, M. peregrinum, M. immunogenum, M. xenopi, M. marinum, M. malmoense, M. marinum, M. mucogenicum, M. nonchromogenicum, M. scrofulaceum, M. simiae, M. smegmatis, M. szulgai, M. terrae, M. terrae complex, M. haemophilum, M. genavense, M. gordonae, M. ulcerans, M. fortuitum or M. fortuitum complex (M. fortuitum and M. chelonae) infections and can be used to treat such infections.

[0006] In one aspect, the present invention provides a system for treating or preventing lung infections. In one embodiment, the system includes a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein the formulation is a dispersion (e.g., a liposome solution or suspension) and the lipid component of the liposome consists of electrically neutral lipids, and a nebulizer that generates an aerosol of the pharmaceutical formulation at a rate greater than about 0.53 g per minute. In one embodiment, the mass median aerodynamic diameter (MMAD) of the aerosol is less than about 4.2 μm when measured by an Anderson Cascade Impactor (ACI), about 3.2 μm to about 4.2 μm when measured by the ACI, or less than about 4.9 μm when measured by a Next Generation Impactor (NGI), about 4.4 μm to about 4.9 μm when measured by the NGI.

[0007] In another embodiment, a system for treating or preventing lung infections includes a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein the formulation is a dispersion (e.g., a liposome solution or suspension) and the lipid component of the liposome consists of electrically neutral lipids, and a nebulizer that generates an aerosol of the pharmaceutical formulation at a rate greater than about 0.53 g per minute. The fine particle fraction (FPF) of the aerosol is greater than or equal to about 64% when measured by an Anderson Cascade Impactor (ACI) or greater than or equal to about 51% when measured by a Next Generation Impactor (NGI).

[0008] In one embodiment, the system provided herein includes a pharmaceutical formulation containing an aminoglycoside. In a further embodiment, the aminoglycoside is amikacin, apramycin, albekacin, astromicin, capreomycin, dibekacin, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamycin or combinations thereof. In yet another embodiment, the aminoglycoside is amikacin. In another embodiment, the aminoglycoside is selected from the aminoglycosides described in Table A below or combinations thereof.

Table A

[0009] The pharmaceutical formulation provided herein is a dispersion of liposomes (i.e., a liposome dispersion or an aqueous liposome dispersion, which may be either a liposome solution or a liposome suspension). In one embodiment, the lipid component of the liposome consists essentially of one or more electrically neutral lipids. In a further embodiment, the electrically neutral lipids include phospholipids and sterols. In a further embodiment, the phospholipid is dipalmitoyl phosphatidylcholine (DPPC) and the sterol is cholesterol.

[0010] In one embodiment, the lipid-to-drug ratio in the aminoglycoside pharmaceutical formulation (aminoglycoside liposome solution or suspension) is about 2:1, about 2:1 or less, about 1:1, about 1:1 or less, or about 0.7:1.

[0011] In one embodiment, the aerosolized aminoglycoside formulation has an aerosol droplet diameter of about 1 μm to about 3.8 μm, about 1.0 μm to 4.8 μm, about 3.8 μm to about 4.8 μm, or about 4.0 μm to about 4.5 μm upon atomization. In a further embodiment, the aminoglycoside is amikacin. In yet another embodiment, the amikacin is amikacin sulfate.

[0012] In one embodiment, about 70% to about 100% of the aminoglycoside present in the formulation is liposome-complexed (e.g., encapsulated in multiple liposomes) prior to atomization. In a further embodiment, the aminoglycoside is selected from the aminoglycosides presented in Table A. In a further embodiment, the aminoglycoside is amikacin. In yet another embodiment, about 80% to about 100% of the amikacin is liposome-complexed or about 80% to about 100% of the amikacin is encapsulated in multiple liposomes. In another embodiment, about 80% to about 100%, about 80% to about 99%, about 90% to about 100%, 90% to about 99%, or about 95% to about 99% of the aminoglycoside present in the formulation prior to atomization is liposome-complexed prior to atomization.

[0013] In one embodiment, the percentage of liposome-complexed (also referred to herein as "liposome association") aminoglycoside after atomization is about 50% to about 80%, about 50% to about 75%, about 50% to about 70%, about 55% to about 75%, or about 60% to about 70%. In a further embodiment, the aminoglycoside is selected from the aminoglycosides presented in Table A. In a further embodiment, the aminoglycoside is amikacin. In yet another embodiment, the amikacin is amikacin sulfate.

[0014] In another aspect, the present invention provides a method for treating or preventing pulmonary infections. In one embodiment, the pulmonary infection is a pulmonary infection caused by Gram-negative bacteria (also referred to herein as Gram-negative bacterial infection). In one embodiment, the pulmonary infection is a Pseudomonas infection, such as a Pseudomonas aeruginosa infection. In another embodiment, the pulmonary infection is caused by one of the species of the genus Pseudomonas presented in Table B below. In one embodiment, the patient is treated for mycobacterial pulmonary infection by one of the systems provided herein. In a further embodiment, the mycobacterial pulmonary infection is a non-tuberculous mycobacterial pulmonary infection, a Mycobacterium abscessus pulmonary infection or a Mycobacterium avium complex pulmonary infection. In one or more of the foregoing embodiments, the patient is a patient with cystic fibrosis.

[0015] In one embodiment, a patient having cystic fibrosis undergoes treatment of a lung infection by one of the systems provided herein. In a further embodiment, the lung infection is caused by Mycobacterium abscessus, Mycobacterium avium complex, or P. aeruginosa. In another embodiment, the lung infection is caused by a nontuberculous mycobacterium selected from M. avium, M. avium subsp. hominissuis (MAH), M. abscessus, M. chelonae, M. bolletii, M. kansasii, M. ulcerans, M. avium, Mycobacterium avium complex (MAC) (M. avium and M. intracellulare), M. conspicuum, M. kansasii, M. peregrinum, M. immunogenum, M. xenopi, M. marinum, M. malmoense, M. marinum, M. mucogenicum, M. nonchromogenicum, M. scrofulaceum, M. simiae, M. smegmatis, M. szulgai, M. terrae, M. terrae complex, M. haemophilum, M. genavense, M. asiaticum, M. shimoidei, M. gordonae, M. nonchromogenicum, M. triplex, M. lentiflavum, M. celatum, M. fortuitum, M. fortuitum complex (M. fortuitum and M. chelonae) or combinations thereof.

[0016] In another aspect, a method for treating or preventing a patient's pulmonary infection is provided. In one embodiment, the method includes aerosolizing a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion of liposomes (e.g., a liposome solution or a liposome suspension) and is aerosolized at a rate greater than about 0.53 grams per minute. The method further includes administering the aerosolized pharmaceutical formulation to the patient's lungs, wherein the aerosolized pharmaceutical formulation comprises a mixture of free aminoglycoside and liposome-complexed aminoglycoside, and the lipid component of the liposomes consists of electrically neutral lipids. In a further embodiment, the mass median aerodynamic diameter (MMAD) of the aerosol is from about 1.0 μm to about 4.2 μm as measured by ACI. In any one of the foregoing embodiments, the MMAD of the aerosol is from about 3.2 μm to about 4.2 μm as measured by ACI. In any one of the foregoing embodiments, the MMAD of the aerosol is from about 1.0 μm to about 4.9 μm as measured by NGI. In any one of the foregoing embodiments, the MMAD of the aerosol is from about 4.4 μm to about 4.9 μm as measured by NGI.

[0017] In one embodiment, the method includes aerosolizing a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate greater than about 0.53 grams per minute. The method further includes administering the aerosolized pharmaceutical formulation to the patient's lungs, wherein the aerosolized pharmaceutical formulation comprises a mixture of free aminoglycoside and liposome-complexed aminoglycoside (e.g., aminoglycoside encapsulated in liposomes), and the liposome component of the formulation consists of electrically neutral lipids. In yet another embodiment, the fine particle fraction (FPF) of the aerosol is greater than or equal to about 64% as measured by ACI or greater than or equal to about 51% as measured by NGI.

[0018] In another aspect, a liposome-complexed aminoglycoside aerosol (e.g., liposome-complexed aminoglycoside) is provided. In one embodiment, the aerosol comprises an aminoglycoside and a plurality of liposomes comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposome-complexed, and the aerosol is produced at a rate greater than about 0.53 grams per minute. In a further embodiment, about 65% to about 75% of the aminoglycoside is liposome-complexed, and the aerosol is produced at a rate greater than about 0.53 grams per minute. In any one of the foregoing embodiments, the aerosol is produced at a rate greater than about 0.54 grams per minute. In any one of the foregoing embodiments, the aerosol is produced at a rate greater than about 0.55 grams per minute. In any one of the foregoing embodiments, the aminoglycoside is selected from the aminoglycosides presented in Table A.

[0019] In one embodiment, the MMAD of the liposome-complexed aminoglycoside aerosol is about 3.2 μm to about 4.2 μm as measured by ACI, or about 4.4 μm to about 4.9 μm as measured by NGI. In a further embodiment, the aerosol comprises an aminoglycoside and a plurality of liposomes comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposome-complexed (e.g., encapsulated in a plurality of liposomes), and the liposome aminoglycoside aerosol is produced at a rate greater than about 0.53 grams per minute. In a further embodiment, the aminoglycoside is selected from the aminoglycosides presented in Table A.

[0020] In one embodiment, the fine particle fraction (FPF) of the lipid-complexed aminoglycoside aerosol is greater than or equal to about 64% when measured by an Andersen Cascade Impactor (ACI), or greater than or equal to about 51% when measured by a Next Generation Impactor (NGI). In a further embodiment, the aerosol comprises an aminoglycoside and a plurality of liposomes comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposome-complexed, for example encapsulated within the plurality of liposomes, and the liposome aminoglycoside aerosol is produced at a rate greater than about 0.53 grams per minute. In any one of the foregoing embodiments, the aerosol is produced at a rate greater than about 0.54 grams per minute. In any one of the foregoing embodiments, the aerosol is produced at a rate greater than about 0.55 grams per minute. In any of the foregoing embodiments, the aminoglycoside is selected from the aminoglycosides presented in Table A.

[0021] In one embodiment, the aerosol comprises an aminoglycoside and a plurality of liposomes comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposome-complexed. In a further embodiment, about 65% to about 75% of the aminoglycoside is encapsulated within the plurality of liposomes. In a further embodiment, the aerosol is produced at a rate greater than about 0.53 grams per minute, greater than about 0.54 grams per minute, or greater than about 0.55 grams per minute. In a further embodiment, the aminoglycoside is amikacin (such as amikacin sulfate).

[0022] In one embodiment, the concentration of the aminoglycoside in the liposome-complexed aminoglycoside is about 50 mg / mL or more. In a further embodiment, the concentration of the aminoglycoside in the liposome-complexed aminoglycoside is about 60 mg / mL or more. In a further embodiment, the concentration of the aminoglycoside in the liposome-complexed aminoglycoside is about 70 mg / mL or more, for example, about 70 mg / mL to about 75 mg / mL. In a further embodiment, the aminoglycoside is selected from the aminoglycosides presented in Table A. In yet another embodiment, the aminoglycoside is amikacin (e.g., amikacin sulfate). In embodiments of the present invention, for example, the following items are provided. (Item 1) A system for treating or preventing pulmonary infections in a patient, the system comprising: (a) a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, the formulation being an aqueous dispersion, and the lipid component of the liposome consisting of electrically neutral lipids; and (b) a nebulizer for generating an aerosol of the pharmaceutical formulation at a rate greater than about 0.53 g per minute, wherein the aerodynamic mass median diameter (MMAD) of the aerosol is less than about 4.2 μm when measured by an Andersen cascade impactor (ACI) or less than about 4.9 μm when measured by a next generation impactor (NGI). (Item 2) A system for treating or preventing pulmonary infections, the system comprising: (a) a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, the formulation being an aqueous dispersion, and the lipid component of the liposome consisting of electrically neutral lipids; and (b) a nebulizer for generating an aerosol of the pharmaceutical formulation at a rate greater than about 0.53 g per minute, A system that includes, and has a fine particle fraction (FPF) of the aerosol that is higher than or equal to about 64% when measured by ACI, or higher than or equal to about 51% when measured by NGI. (Item 3) The system according to item 1 or 2, wherein the aminoglycoside is selected from amikacin, apramycin, albekacin, astromicin, capreomycin, dibekacin, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamycin, or a combination thereof. (Item 4) The system according to any one of items 1 to 3, wherein the aminoglycoside is amikacin. (Item 5) The system according to any one of items 1 to 4, wherein the aminoglycoside is amikacin sulfate. (Item 6) The system according to any one of items 1 to 5, wherein the liposome contains unilamellar vesicles, multilamellar vesicles, or a mixture thereof. (Item 7) The system according to any one of items 1 to 6, wherein the electrically neutral lipid contains an electrically neutral phospholipid or an electrically neutral phospholipid and a sterol. (Item 8) The system according to any one of items 1 to 7, wherein the electrically neutral lipid contains phosphatidylcholine and a sterol. (Item 9) The system according to any one of items 1 to 8, wherein the electrically neutral lipid contains dipalmitoylphosphatidylcholine (DPPC) and a sterol. (Item 10) The system according to any one of items 1 to 9, wherein the electrically neutral lipid contains DPPC and cholesterol. (Item 11) The system according to any one of items 1 to 10, wherein the aminoglycoside is amikacin, the electrically neutral lipid consists of DPPC and cholesterol, and the liposome contains unilamellar vesicles, multilamellar vesicles, or a mixture thereof. (Item 12) The system according to any one of items 1 to 11, wherein the weight ratio of free aminoglycoside to the liposome-complexed aminoglycoside is from about 1:100 to about 100:1. (Item 13) The system according to any one of items 1 to 12, wherein the weight ratio of free aminoglycoside to the liposome-complexed aminoglycoside is from about 1:10 to about 10:1. (Item 14) The system according to any one of items 1 to 13, wherein the weight ratio of free aminoglycoside to the liposome-complexed aminoglycoside is from about 0.3:1 to about 2:1. (Item 15) The system according to any one of items 1 to 14, wherein the volume of the pharmaceutical preparation is about 8 mL. (Item 16) The system according to any one of items 1 to 15, wherein the aerosol contains about 55% to about 75% of liposome-complexed amikacin. (Item 17) The system according to any one of items 1 to 16, wherein the liposome-complexed aminoglycoside has a MMAD of about 3.2 μm to about 4.2 μm as measured by ACI, or about 4.4 μm to about 4.9 μm as measured by NGI. (Item 18) The system according to any one of items 1 to 17, wherein the liposome-complexed aminoglycoside has a MMAD of about 3.6 μm to about 3.9 μm as measured by ACI, or about 4.5 μm to about 4.8 μm as measured by NGI. (Item 19) The system according to any one of items 1 to 18, wherein the FPF of the aerosolized formulation is higher than or equal to about 64% when measured by ACI, or higher than or equal to about 51% when measured by NGI. (Item 20) The system according to any one of items 1 to 19, wherein the FPF of the aerosolized formulation is from about 64% to about 80% when measured by ACI, or from about 51% to about 65% when measured by NGI. (Item 21) The system according to any one of items 1 to 20, wherein the nebulizer generates an aerosol of the pharmaceutical formulation at a rate exceeding about 0.54 g per minute. (Item 22) The system according to any one of items 1 to 21, wherein the aerosol contains a free aminoglycoside in an amount effective to provide immediate bactericidal activity or immediate antibiotic activity against the pulmonary infection, and a liposome-complexed aminoglycoside in an amount effective to provide sustained bactericidal activity or sustained antibiotic activity against the pulmonary infection. (Item 23) The system according to any one of items 1 to 22, wherein the pharmaceutical formulation contains about 500 mg to about 650 mg of aminoglycoside. (Item 24) The system according to any one of items 1 to 22, wherein the pharmaceutical formulation contains about 550 mg to about 625 mg of aminoglycoside. (Item 25) The system according to any one of items 1 to 22, wherein the pharmaceutical formulation contains about 550 mg to about 600 mg of aminoglycoside. (Item 26) The system according to any one of items 1 to 22, wherein the pharmaceutical formulation contains about 560 mg of aminoglycoside. (Item 27) The system according to any one of items 1 to 22, wherein the pharmaceutical formulation contains about 580 mg of aminoglycoside. (Item 28) The system according to any one of items 1 to 22, wherein the pharmaceutical preparation contains about 590 mg of aminoglycoside. (Item 29) The system according to any one of items 1 to 22, wherein the pharmaceutical preparation contains about 600 mg of aminoglycoside. (Item 30) A method for treating or preventing pulmonary infection in a patient, the method comprising: A step of aerosolizing a pharmaceutical preparation containing a liposome-complexed aminoglycoside, wherein the pharmaceutical preparation is an aqueous dispersion and is aerosolized at a rate greater than about 0.53 grams per minute; and A step of administering the aerosolized pharmaceutical preparation to the lungs of the patient comprising wherein the aerosolized pharmaceutical preparation contains a mixture of free aminoglycoside and liposome-complexed aminoglycoside, the lipid component of the liposome consists of electrically neutral lipids, and the MMAD of the aerosol is less than about 4.2 μm when measured by ACI or less than about 4.9 μm when measured by NGI. (Item 31) The method according to item 30, wherein the MMAD of the aerosol is from about 3.2 μm to about 4.2 μm when measured by ACI or from about 4.4 μm to about 4.9 μm when measured by NGI. (Item 32) A method for providing treatment or prevention of pulmonary infection in a patient, the method comprising: A step of aerosolizing a pharmaceutical preparation containing a liposome-complexed aminoglycoside, wherein the pharmaceutical preparation is an aqueous dispersion and is aerosolized at a rate greater than about 0.53 grams per minute; and A step of administering the aerosolized pharmaceutical preparation to the lungs of the patient comprising wherein the aerosolized pharmaceutical preparation contains a mixture of free aminoglycoside and liposome-complexed aminoglycoside, The lipid component of the liposome consists of electrically neutral lipids, and a method wherein the fine particle fraction (FPF) of the aerosol is higher than or equal to about 64% when measured by ACI, or higher than or equal to about 51% when measured by NGI. (Item 33) The method according to any one of Items 30 to 32, wherein the pharmaceutical preparation is aerosolized at a rate exceeding about 0.55 grams per minute. (Item 34) The method according to any one of Items 30 to 33, wherein the pharmaceutical preparation is aerosolized at a rate exceeding about 0.56 grams per minute. (Item 35) The method according to any one of Items 30 to 34, wherein the pharmaceutical preparation is aerosolized at a rate exceeding about 0.58 grams per minute. (Item 36) The method according to any one of Items 30 to 35, wherein the pharmaceutical preparation is aerosolized at a rate of about 0.60 to 0.80 grams per minute. (Item 37) The method according to any one of Items 30 to 36, wherein the pharmaceutical preparation is aerosolized at a rate of about 0.60 to 0.70 grams per minute. (Item 38) The method according to any one of Items 30 to 37, wherein the pharmaceutical preparation contains about 70 to about 75 mg / mL of amikacin, about 32 to about 35 mg / mL of DPPC, and about 16 to about 17 mg / mL of cholesterol. (Item 39) The method according to any one of Items 30 to 38, wherein the pharmaceutical preparation has a volume of about 8 mL. (Item 40) The method according to any one of items 30 to 39, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, albekacin, astromicin, bekanamycin, bohromycin, bulramycin, capreomycin, dibekacin, dactimicin, etimicin, flamicetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomycin, ribostamycin, sisomicin, rhodostreptomycin, solbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamycin, vertilmicin, or a combination thereof. (Item 41) The method according to any one of items 30 to 40, wherein the aerosolized pharmaceutical formulation is administered once a day in a single administration session. (Item 42) The method according to any one of items 30 to 41, wherein the aminoglycoside is amikacin. (Item 43) The method according to any one of items 30 to 42, wherein the aminoglycoside is amikacin sulfate. (Item 44) The method according to any one of items 30 to 43, wherein the free aminoglycoside is in an amount effective to provide immediate bactericidal activity or immediate antibiotic activity against the nontuberculous mycobacterial infection, and the liposome-complexed aminoglycoside is in an amount effective to provide sustained bactericidal activity or sustained antibiotic activity against the nontuberculous mycobacterial infection. (Item 45) The method according to any one of items 30 to 44, wherein the pharmaceutical formulation contains about 500 mg to about 650 mg of aminoglycoside. (Item 46) The method according to any one of items 30 to 44, wherein the pharmaceutical formulation contains about 550 mg to about 625 mg of aminoglycoside. (Item 47) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains from about 550 mg to about 600 mg of aminoglycoside. (Item 48) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains about 560 mg of aminoglycoside. (Item 49) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains about 580 mg of aminoglycoside. (Item 50) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains about 590 mg of aminoglycoside. (Item 51) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains about 600 mg of aminoglycoside. (Item 52) A step of atomizing an aqueous liposome dispersion of about 8 to about 9 grams of aminoglycoside in less than about 16 minutes to obtain a liposome aminoglycoside aerosol, and A step of delivering the liposome aminoglycoside aerosol to the lungs of a patient by inhalation A method for delivering a liposome aminoglycoside aerosol, comprising: (Item 53) A step of atomizing an aqueous liposome dispersion of about 8 to about 9 grams of aminoglycoside in about 10 to about 15 minutes to obtain a liposome aminoglycoside aerosol, and A method for delivering a liposome aminoglycoside aerosol, comprising a step of delivering the liposome aminoglycoside aerosol to the lungs of a patient by inhalation. (Item 54) The method according to item 52 or 53, wherein the aqueous liposome dispersion of aminoglycoside is atomized in less than about 15 minutes, less than about 14 minutes, less than about 13 minutes, less than about 12 minutes, or less than about 11 minutes. (Item 55) The method according to item 52 or 53, wherein the aqueous liposome dispersion of the aminoglycoside is atomized in about 10 minutes to about 14 minutes, about 10 minutes to about 13 minutes, about 10 minutes to about 12 minutes, about 10 minutes to about 11 minutes, about 11 minutes to about 15 minutes, about 12 minutes to about 15 minutes, about 13 minutes to about 15 minutes or about 14 minutes to about 15 minutes. (Item 56) The method according to any one of items 52 to 55, wherein the liposome aminoglycoside aerosol comprises an aminoglycoside and liposomes composed of DPPC and cholesterol, and about 55% to about 75% of the aminoglycoside is complexed with the liposome. (Item 57) The liposome aminoglycoside aerosol has an MMAD of about 3.2 μm to about 4.2 μm when measured by ACI, or about 4.4 μm to about 4.9 μm when measured by NGI; a GSD of about 1.75 to about 1.80; a FPF that is higher than or equal to about 64% when measured by ACI, or higher than or equal to about 51% when measured by NGI; or an FPD of about 35 to about 41 The method according to any one of items 52 to 56. (Item 58) The method according to any one of items 52 to 57, wherein about 25% to about 35% of the liposome aminoglycoside aerosol deposits in the bronchi and the alveolar region of the patient's lungs. (Item 59) The method according to any one of items 52 to 58, wherein the aqueous liposome dispersion of the aminoglycoside comprises an aminoglycoside and liposomes composed of DPPC and cholesterol, and more than about 95% of the aminoglycoside is encapsulated in the liposome. (Item 60) The method according to any one of items 52 to 59, wherein the aqueous liposome dispersion of the aminoglycoside comprises about 70 to about 75 mg / mL of aminoglycoside, about 32 to about 35 mg / mL of DPPC, and about 16 mg / mL to about 17 mg / mL of cholesterol. (Item 61) The method according to any one of Items 52 to 60, wherein the volume of the aqueous liposome dispersion is about 8 mL. (Item 62) The method according to any one of Items 52 to 61, wherein the aminoglycoside is amikacin. (Item 63) The method according to any one of Items 52 to 62, wherein the aminoglycoside is amikacin sulfate. (Item 64) The method according to any one of Items 52 to 63, wherein about 500 mg to about 650 mg of aminoglycoside is delivered to the lungs of the patient by inhalation. (Item 65) The method according to any one of Items 52 to 63, wherein about 550 mg to about 625 mg of aminoglycoside is delivered to the lungs of the patient by inhalation. (Item 66) The method according to any one of Items 52 to 63, wherein about 550 mg to about 600 mg of aminoglycoside is delivered to the lungs of the patient by inhalation. (Item 67) The method according to any one of Items 52 to 63, wherein about 560 mg of aminoglycoside is delivered to the lungs of the patient by inhalation. (Item 68) The method according to any one of Items 52 to 63, wherein about 580 mg of aminoglycoside is delivered to the lungs of the patient by inhalation. (Item 69) The method according to any one of Items 52 to 63, wherein about 590 mg of aminoglycoside is delivered to the lungs of the patient by inhalation. (Item 70) The method according to any one of Items 52 to 63, wherein about 600 mg of aminoglycoside is delivered to the lungs of the patient by inhalation. (Item 71) A liposomal aminoglycoside aerosol comprising an aminoglycoside and liposomes containing DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is complexed with the liposomes, and the liposomal aminoglycoside aerosol is produced at a rate exceeding about 0.53 grams per minute. (Item 72) The liposomal aminoglycoside aerosol according to item 71, produced at a rate higher than about 0.54 grams per minute or equal to about 0.54 grams per minute, or higher than about 0.55 grams per minute or equal to about 0.55 grams per minute, or higher than about 0.60 grams per minute or equal to about 0.60 grams per minute. (Item 73) The liposomal aminoglycoside aerosol according to item 71 or 72, produced at a rate of about 0.60 to about 0.70 grams per minute. (Item 74) The liposomal aminoglycoside aerosol according to any one of items 71 to 73, wherein about 500 mg or about 560 mg of amikacin is provided in a single administration session. (Item 75) The liposomal aminoglycoside aerosol according to item 74, wherein the administration session is once a day. (Item 76) The liposomal aminoglycoside aerosol according to any one of items 71 to 75, wherein the MMAD of the aerosol is less than about 4.2 μm when measured by ACI, or less than about 4.9 μm when measured by NGI. (Item 77) The liposomal aminoglycoside aerosol according to any one of items 71 to 76, wherein the MMAD of the aerosol is about 1.0 μm to about 4.2 μm when measured by ACI, about 2.0 μm to about 4.2 μm when measured by ACI, about 3.2 μm to about 4.2 μm when measured by ACI, about 1.0 μm to about 4.9 μm when measured by NGI, about 2.0 μm to about 4.9 μm when measured by NGI, or about 4.4 μm to about 4.9 μm when measured by NGI. (Item 78) The liposomal aminoglycoside aerosol according to any one of items 71 to 77, wherein the fine particle fraction (FPF) of the aerosol is higher than or equal to about 64% when measured by ACI, or higher than or equal to about 51% when measured by NGI. (Item 79) The liposomal aminoglycoside aerosol according to any one of items 71 to 78, wherein the aerosol contains liposome-complexed amikacin that is higher than or equal to about 50%. (Item 80) The liposomal aminoglycoside aerosol according to any one of items 71 to 79, wherein the aerosol contains liposome-complexed amikacin that is higher than or equal to about 60%. (Item 81) The liposomal aminoglycoside aerosol according to any one of items 71 to 80, wherein the aerosol contains 55% to 85% of liposome-complexed amikacin. (Item 82) The liposomal aminoglycoside aerosol according to any one of items 71 to 81, wherein the aerosol contains 55% to 75% of liposome-complexed amikacin. (Item 83) The liposomal aminoglycoside aerosol according to any one of items 71 to 82, wherein the liposome contains unilamellar vesicles, multilamellar vesicles, or a mixture thereof. (Item 84) The liposomal aminoglycoside aerosol according to any one of items 71 to 83, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, albekacin, astromicin, bekanamycin, bohromycin, bulramycin, capreomycin, dibekacin, dactimicin, etimicin, flamicetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomycin, ribostamycin, sisomicin, rhodostreptomycin, solbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamycin, vertilmicin, or a combination thereof. (Item 85) A liposomal aminoglycoside aerosol comprising a plurality of liposomes containing an aminoglycoside, DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is complexed with the liposomes, and the plurality of liposomes have a diameter of about 245 nm to about 290 nm as measured by light scattering. (Item 86) The liposomal aminoglycoside aerosol according to item 85, which is produced at a rate exceeding about 0.53 grams per minute. (Item 87) The liposomal aminoglycoside aerosol according to item 85 or 86, wherein about 560 mg of amikacin is provided in one administration session. (Item 88) The liposomal aminoglycoside aerosol according to any one of items 85 to 87, wherein the administration session is once a day. (Item 89) When the MMAD of the aerosol is measured by ACI, it is about 1.0 μm to about 4.2 μm, when measured by ACI, it is about 2.0 μm to about 4.2 μm, when measured by ACI, it is about 3.2 μm to about 4.2 μm, when measured by NGI, it is about 1.0 μm to about 4.9 μm, when measured by NGI, it is about 2.0 μm to about 4.9 μm, when measured by NGI, it is about 4.4 μm to about 4.9 μm, the liposomal aminoglycoside aerosol according to any one of items 85 to 88. (Item 90) When the MMAD of the aerosol is measured by ACI, it is about 3.2 μm to about 4.2 μm, or when measured by NGI, it is about 4.4 μm to about 4.9 μm, the liposomal aminoglycoside aerosol according to any one of items 85 to 89. (Item 91) When the FPF of the aerosol is measured by ACI, it is higher than about 64% or equal to about 64%, or when measured by NGI, it is higher than about 51% or equal to about 51%, the liposomal aminoglycoside aerosol according to any one of items 85 to 90. (Item 92) The aerosol contains more than about 55% of liposome-complexed amikacin, more than about 60% of liposome-complexed amikacin, more than about 65% of liposome-complexed amikacin, or more than about 70% of liposome-complexed amikacin, the liposomal aminoglycoside aerosol according to any one of items 85 to 91. (Item 93) The aerosol contains about 55% to about 75% of liposome-complexed amikacin, the liposomal aminoglycoside aerosol according to any one of items 85 to 92. (Item 94) The liposome contains unilamellar vesicles, multilamellar vesicles, or a mixture thereof, the liposomal aminoglycoside aerosol according to any one of items 85 to 93. (Item 95) The liposomal aminoglycoside aerosol according to any one of items 85 to 94, wherein the aminoglycoside is selected from amikacin, apramycin, albekacin, astromicin, capreomycin, dibekacin, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamycin, or a combination thereof. (Item 96) The liposomal aminoglycoside aerosol according to any one of items 71 to 95, which is generated at a rate of about 0.55 to about 0.70 grams per minute, or at a rate of about 0.60 to about 0.70 grams per minute, or at a rate of about 0.65 to about 0.70 grams per minute. (Item 97) The liposomal aminoglycoside aerosol according to any one of items 85 to 96, wherein the average liposome size is about 265 nm when measured by light scattering. (Item 98) The method according to any one of items 30 to 51, wherein the patient has cystic fibrosis. (Item 99) The method according to any one of items 30 to 51 and 98, wherein the pulmonary infection is a nontuberculous mycobacterial infection. (Item 100) The method according to any one of items 30 to 51 and 98, wherein the pulmonary infection is a Pseudomonas infection. (Item 101) The method according to any one of items 30 to 51 and 98, wherein the pulmonary infection is a Burkholderia infection. (Item 102) The method according to item 100, wherein the Pseudomonas infection is a Pseudomonas aeruginosa infection. (Item 104) The method according to item 101, wherein the Burkholderia infection is caused by B. pseudomallei, B. cepacia, B. cepacia complex, B. dolosa, B. fungorum, B. gladioli, B. multivorans, B. vietnamiensis, B. pseudomallei, B. ambifaria, B. andropogonis, B. anthina, B. brasilensis, B. caledonica, B. caribensis or B. caryophylli. (Item 105) The method according to item 99, wherein the non-tuberculous mycobacterial infection is M. avium. (Item 106) The method according to item 105, wherein the M. avium infection is Mycobacterium avium subsp. hominissuis infection. (Item 107) The method according to item 99, wherein the non-tuberculous mycobacterial infection is Mycobacterium abscessus infection. (Item 108) The method according to item 99, wherein the non-tuberculous mycobacterial infection is Mycobacterium avium complex (M. avium and M. intracellulare). (Item 109) The method according to item 99, wherein the non-tuberculous mycobacterial infection is selected from M. avium, M. avium subsp. hominissuis (MAH), M. abscessus, M. chelonae, M. bolletii, M. kansasii, M. ulcerans, M. avium, M. avium complex (MAC) (M. avium and M. intracellulare), M. conspicuum, M. kansasii, M. peregrinum, M. immunogenum, M. xenopi, M. marinum, M. malmoense, M. marinum, M. mucogenicum, M. nonchromogenicum, M. scrofulaceum, M. simiae, M. smegmatis, M. szulgai, M. terrae, M. terrae complex, M. haemophilum, M. genavense, M. asiaticum, M. shimoidei, M. gordonae, M. nonchromogenicum, M. triplex, M. lentiflavum, M. celatum, M. fortuitum, M. fortuitum complex (M. fortuitum and M. chelonae), or a combination thereof. (Item 110) The system according to any one of items 1 to 29, wherein the concentration of the aminoglycoside is about 50 mg / mL or more, or about 60 mg / mL or more, or about 70 mg / mL or more. (Item 111) The system according to any one of items 1 to 29, wherein the concentration of the aminoglycoside is about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 76 mg / mL, about 77 mg / mL, about 78 mg / mL, or about 79 mg / mL. (Item 112) The system according to any one of items 1 to 29, wherein the concentration of the aminoglycoside is about 60 mg / mL to about 80 mg / mL. (Item 113) The system according to any one of items 110 to 112, wherein the aminoglycoside is amikacin. (Item 114) The system according to item 113, wherein the aminoglycoside is amikacin sulfate. (Item 115) The method according to any one of items 30 to 70 and 98 to 109, wherein the concentration of the aminoglycoside is about 50 mg / mL or more, or about 60 mg / mL or more, or about 70 mg / mL or more. (Item 116) The method according to any one of items 30 to 70 and 98 to 109, wherein the concentration of the aminoglycoside is about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 76 mg / mL, about 77 mg / mL, about 78 mg / mL or about 79 mg / mL. (Item 117) The method according to any one of items 30 to 70 and 98 to 109, wherein the concentration of the aminoglycoside is about 60 mg / mL to about 80 mg / mL. (Item 118) The method according to any one of items 30 to 70 and 98 to 109, wherein the concentration of the aminoglycoside is about 70 mg / mL to about 80 mg / mL. (Item 119) The method according to any one of items 116 to 118, wherein the aminoglycoside is amikacin. (Item 120) The method according to item 119, wherein the aminoglycoside is amikacin sulfate. (Item 121) The liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is about 50 mg / mL or more, or about 60 mg / mL or more, or about 70 mg / mL or more. (Item 122) The liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 76 mg / mL, about 77 mg / mL, about 78 mg / mL or about 79 mg / mL. (Item 123) The liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is about 60 mg / mL to about 80 mg / mL. (Item 124) The liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is about 70 mg / mL to about 80 mg / mL. (Item 125) The liposomal aminoglycoside aerosol according to any one of items 122 to 124, wherein the aminoglycoside is amikacin. (Item 126) The liposomal aminoglycoside aerosol according to item 125, wherein the aminoglycoside is amikacin sulfate. (Item 127) The system according to any one of items 1 to 29, wherein the lung infection is a non-tuberculous mycobacterial lung infection. (Item 128) The system according to any one of items 1 to 29, wherein the lung infection is a Pseudomonas lung infection. (Item 129) The system according to any one of items 1 to 29, wherein the lung infection is a Burkholderia lung infection. (Item 130) The system according to item 128, wherein the Pseudomonas infection is a Pseudomonas aeruginosa lung infection. (Item 131) The system according to item 129, wherein the Burkholderia lung infection is a lung infection caused by B. pseudomallei, B. cepacia, B. cepacia complex, B. dolosa, B. fungorum, B. gladioli, B. multivorans, B. vietnamiensis, B. pseudomallei, B. ambifaria, B. andropogonis, B. anthina, B. brasilensis, B. caledonica, B. caribensis or B. caryophylli. (Item 132) The system according to item 127, wherein the non-tuberculous mycobacterial lung infection is a lung infection caused by M. avium, M. abscessus, M. chelonae, M. bolletii, M. kansasii, M. ulcerans, M. avium, M. avium complex (MAC) (M. avium and M. intracellulare), M. conspicuum, M. kansasii, M. peregrinum, M. immunogenum, M. xenopi, M. marinum, M. malmoense, M. marinum, M. mucogenicum, M. nonchromogenicum, M. scrofulaceum, M. simiae, M. smegmatis, M. szulgai, M. terrae, M. terrae complex, M. haemophilum, M. genavense, M. asiaticum, M. shimoidei, M. gordonae, M. nonchromogenicum, M. triplex, M. lentiflavum, M. celatum, M. fortuitum, M. fortuitum complex (M. fortuitum and M. chelonae), or a combination thereof. (Item 133) The system according to item 132, wherein the non-tuberculous mycobacterial lung infection is an M. abscessus lung infection. (Item 134) The system according to item 132, wherein the non-tuberculous mycobacterial lung infection is an M. avium lung infection. (Item 135) The system according to item 134, wherein the non-tuberculous mycobacterial pulmonary infection is M. avium subsp. hominissuis pulmonary infection. (Item 137) The method according to any one of items 30 to 39, 41 and 44 to 70, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, albekacin, astromicin, bekanamycin, bohoromycin, bulramycin, capreomycin, dibekacin, dactimicin, etimicin, framycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomycin, ribostamycin, sisomicin, rhodostreptomycin, solbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamycin, vertilmicin, or a combination thereof. (Item 138) The system according to any one of items 1 to 3, 6 to 29 and 110 to 114, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, albekacin, astromicin, bekanamycin, bohoromycin, bulramycin, capreomycin, dibekacin, dactimicin, etimicin, framycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomycin, ribostamycin, sisomicin, rhodostreptomycin, solbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamycin, vertilmicin, or a combination thereof. (Item 139) The system according to any one of items 1 to 29, 110 to 114 and 138, wherein the aqueous dispersion is an aqueous suspension of liposome-complexed aminoglycoside. (Item 140) The system according to any one of items 1 to 29, 110 to 114, and 138, wherein the aqueous dispersion is an aqueous solution of a liposome-complexed aminoglycoside. (Item 141) The method according to any one of items 30 to 70, 115 to 120, and 137, wherein the aqueous dispersion is an aqueous suspension of a liposome-complexed aminoglycoside. (Item 142) The method according to any one of items 30 to 70, 115 to 120, and 137, wherein the aqueous dispersion is an aqueous solution of a liposome-complexed aminoglycoside. (Item 143) The method according to any one of items 30 to 51 and 98, wherein the lung infection is associated with bronchiectasis. (Item 144) The method according to item 143, wherein the aqueous dispersion is an aqueous suspension of a liposome-complexed aminoglycoside. (Item 145) The method according to item 143, wherein the aqueous dispersion is an aqueous solution of a liposome-complexed aminoglycoside. (Item 146) The system according to any one of items 1 to 29, wherein the lung infection is associated with bronchiectasis.

Brief Description of the Drawings

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[0037] DETAILED DESCRIPTION OF THE INVENTION One aspect of the invention described herein is directed to a system for administering an aminoglycoside pharmaceutical formulation to a subject's lung, for example, to treat a lung disorder.

[0038] The term "treating" includes (1) preventing or delaying the onset of clinical symptoms of a condition, disorder or state in a subject who is suffering from or may be predisposed to the condition, disorder or state but has not yet experienced or exhibited clinical or pre-clinical symptoms of the condition, disorder or state; (2) inhibiting the condition, disorder or state (i.e., suppressing, reducing or delaying the occurrence of at least one clinical or pre-clinical symptom of the disease or, in the case of maintenance treatment, its recurrence); and / or (3) alleviating the state (i.e., causing at least a regression of the condition, disorder or state, or at least one of its clinical or pre-clinical symptoms). The benefit to the subject being treated is statistically significant or at least perceptible to the subject or the physician.

[0039] In one embodiment, the systems and formulations provided herein can treat lung infections caused by the following bacteria: Pseudomonas (e.g., P. aeruginosa, P. paucimobilis, P. putida, P. fluorescens, and P. acidovorans), Burkholderia (e.g., B. pseudomallei, B. cepacia, B. cepacia complex, B. dolosa, B. fungorum, B. gladioli, B. multivorans, B. vietnamiensis, B. pseudomallei, B. ambifaria, B. andropogonis, B. anthina, B. brasilensis, B. caledonica, B. caribensis, B. caryophylli), Staphylococcus (e.g., S. aureus, S. auricularis, S. carnosus, S. epidermidis, S. lugdunensis), methicillin-resistant Staphylococcus aureus (MRSA), Streptococcus (e.g., Streptococcus pneumoniae), Escherichia coli, Klebsiella, Enterobacter, Serratia, Haemophilus, Yersinia pestis, Mycobacterium (e.g., nontuberculous mycobacteria).

[0040] In one embodiment, a patient receives treatment for nontuberculous mycobacterial lung infection by one of the systems provided herein. In a further embodiment, the nontuberculous mycobacterial lung infection is a refractory nontuberculous mycobacterial lung infection.

[0041] In one embodiment, the systems provided herein are used for the treatment of a patient having a lung infection caused by Pseudomonas. In a further embodiment, the lung infection is caused by a species of the genus Pseudomonas selected from the species presented in Table B below.

Table B-1

Table B-2

[0042] In one embodiment, nontuberculous mycobacterial pulmonary infection is selected from M. avium, M. avium subsp. hominissuis (MAH), M. abscessus, M. chelonae, M. bolletii, M. kansasii, M. ulcerans, M. avium, M. avium complex (MAC) (M. avium and M. intracellulare), M. conspicuum, M. kansasii, M. peregrinum, M. immunogenum, M. xenopi, M. marinum, M. malmoense, M. marinum, M. mucogenicum, M. nonchromogenicum, M. scrofulaceum, M. simiae, M. smegmatis, M. szulgai, M. terrae, M. terrae complex, M. haemophilum, M. genavense, M. asiaticum, M. shimoidei, M. gordonae, M. nonchromogenicum, M. triplex, M. lentiflavum, M. celatum, M. fortuitum, M. fortuitum complex (M. fortuitum and M. chelonae) or combinations thereof. In a further embodiment, nontuberculous mycobacterial pulmonary infection is M. abscessus or M. avium. In a further embodiment, M. avium infection is M. avium subsp. hominissuis. In one embodiment, nontuberculous mycobacterial pulmonary infection is refractory nontuberculous mycobacterial pulmonary infection.

[0043] In another embodiment, a patient with cystic fibrosis undergoes treatment of a bacterial infection by one of the systems provided herein. In a further embodiment, the bacterial infection is a lung infection by Pseudomonas aeruginosa. In yet another embodiment, the patient undergoes treatment of a lung infection associated with bronchiectasis by one of the systems provided herein.

[0044] As used herein, "prevention" can mean the complete prevention of an infectious disease or disorder, or the prevention of the occurrence of the symptoms of that infectious disease or disorder; the delay in the onset of an infectious disease or disorder or its symptoms; or a reduction in the severity of an infectious disease or disorder or its symptoms that occur later.

[0045] The term "antibacterial" is recognized in the art and refers to the ability of the compounds of the present invention to prevent, inhibit or destroy the microbial growth of bacteria. Examples of bacteria are presented above.

[0046] The term "antimicrobial" is recognized in the art and refers to the ability of the aminoglycoside compounds of the present invention to prevent, inhibit, delay or destroy the growth of microorganisms such as bacteria, fungi, protozoa and viruses.

[0047] "Effective amount" means an amount of an aminoglycoside (e.g., amikacin) used in the present invention that is sufficient to produce a desired therapeutic response. The formulations of the effective amount provided herein include both free aminoglycoside and liposome-complexed aminoglycoside. For example, liposome-complexed aminoglycoside, in one embodiment, includes aminoglycoside encapsulated in liposomes, or aminoglycoside complexed with liposomes, or a combination thereof.

[0048] In one embodiment, the aminoglycoside is selected from amikacin, apramycin, albekacin, astromicin, capreomycin, dibekacin, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin or verdamycin. In another embodiment, the aminoglycoside is selected from the aminoglycosides described in Table C below.

Table C

[0049] In one embodiment, the aminoglycoside is an aminoglycoside free base, or a salt, solvate, or other non-covalent derivative thereof. In a further embodiment, the aminoglycoside is amikacin. Suitable aminoglycosides for use in the pharmaceutical formulations of the present invention include pharmaceutically acceptable addition salts and complexes of the drug. If the compound can have one or more chiral centers, unless otherwise specified, the present invention includes each unique racemic compound, as well as each unique non-racemic compound. If the active agent has an unsaturated carbon-carbon double bond, both the cis (Z) isomer and the trans (E) isomer are included within the scope of the present invention. If the active agent exists in a tautomeric form such as a keto-enol tautomer, each tautomeric form is considered to be included in the present invention. Amikacin is present in the pharmaceutical formulation, in one embodiment, as amikacin base, or an amikacin salt, such as amikacin sulfate, i.e., amikacin disulfate. In one embodiment, one or more combinations of the above aminoglycosides are used in the formulations, systems and methods described herein. In a further embodiment, the combination includes amikacin.

[0050] A treatment response can be any response that a user (e.g., a clinician) would recognize as an effective response to a treatment. A treatment response will generally be a reduction, inhibition, delay, or prevention of the growth or proliferation of one or more of the bacteria described above, or the killing of one or more of the bacteria described above. A treatment response can also be reflected in an improvement in lung function, such as forced expiratory volume in one second (FEV1). Furthermore, based on the evaluation of the treatment response, determining an appropriate treatment duration, an appropriate dosage, and any potential combination treatments is within the skill of those in the art.

[0051] A "liposome dispersion" refers to a solution or suspension containing a plurality of liposomes.

[0052] As used herein, an "aerosol" is a gas-phase suspension of liquid particles. The aerosols provided herein contain particles of the liposome dispersion.

[0053] A "nebulizer" or "aerosol generator" is a device that converts a liquid into an aerosol of a size that can be inhaled into the airways. Pneumatic nebulizers, ultrasonic nebulizers, electronic nebulizers, such as passive electronic mesh nebulizers, active electronic mesh nebulizers, and vibrating mesh nebulizers, are suitable for use with the present invention if the particular nebulizer releases an aerosol of the required properties at the required delivery rate.

[0054] The process of converting a bulk liquid into small droplets with air is called atomization. For the operation of a pneumatic nebulizer, a pressurized gas supply is required as the driving force for liquid atomization. An ultrasonic nebulizer converts a liquid into breathable droplets using electricity introduced by a piezoelectric element in a liquid reservoir. Various types of nebulizers are described in Respiratory Care, Vol. 45, No. 6, pp. 609 - 622 (2000), the disclosure of which is hereby incorporated by reference in its entirety. The terms "nebulizer" and "aerosol generator" are used interchangeably throughout this specification. In the literature, the terms "inhalation device", "inhalation system" and "atomizer" are also used interchangeably with the terms "nebulizer" and "aerosol generator".

[0055] As used herein, "Fine particle fraction" or "FPF" refers to the fraction of aerosol having a particle size less than 5 μm when measured by cascade impaction. FPF is usually expressed as a percentage.

[0056] "Mass median diameter" or "MMD" is determined by laser diffraction or impacter measurement and is the average particle diameter on a mass basis.

[0057] "Aerodynamic mass median diameter" or "MMAD" is standardized with respect to the aerodynamic separation of aqueous aerosol droplets and is determined by impacter measurement, such as an Andersen cascade impacter (ACI) or a next generation impacter (NGI). The gas flow rate is, in one embodiment, 28 liters per minute for an Andersen cascade impacter (ACI) and 15 liters per minute for a next generation impacter (NGI). "Geometric standard deviation" or "GSD" is a measure of the spread of the aerodynamic particle size distribution.

[0058] In one embodiment, the present invention provides a system for treating or preventing pulmonary infections. The treatment is achieved via delivery of an aminoglycoside formulation by inhalation via nebulization. In one embodiment, the pharmaceutical formulation comprises an aminoglycoside agent, such as an aminoglycoside.

[0059] The pharmaceutical formulation provided herein is a liposome dispersion. Specifically stated, the pharmaceutical formulation is a dispersion comprising "liposome-complexed aminoglycoside" or "aminoglycoside encapsulated in liposomes". "Liposome-complexed aminoglycoside" encompasses embodiments in which an aminoglycoside (or combination of aminoglycosides) is encapsulated in liposomes, and includes any form of aminoglycoside composition in which at least about 1% by weight of the aminoglycoside is associated with the liposomes, either as part of a complex with the liposomes, or as liposomes in which the aminoglycoside is in the aqueous phase or the hydrophobic bilayer phase, or present in the interfacial headgroup region of the liposome bilayer.

[0060] In one embodiment, the lipid component of the liposome comprises an electrically neutral lipid, a positively charged lipid, a negatively charged lipid, or a combination thereof. In another embodiment, the lipid component comprises an electrically neutral lipid. In a further embodiment, the lipid component consists essentially of an electrically neutral lipid. In yet another embodiment, the lipid component consists of an electrically neutral lipid, such as a sterol and a phospholipid.

[0061] As presented above, liposome-complexed aminoglycoside embodiments include embodiments in which the aminoglycoside is encapsulated within the liposome. Additionally, liposome-complexed aminoglycoside refers to any composition, solution or suspension in which at least about 1% by weight of the aminoglycoside is associated with the lipid, either as part of a complex with the liposome, or where the aminoglycoside is in the aqueous or hydrophobic bilayer phase, or may be present in the interfacial head group region of the liposome bilayer as a liposome. In one embodiment, prior to nebulization, at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% of the aminoglycoside in the formulation is so associated. The association is, in one embodiment, measured by filtration separation such that the lipid and lipid-associated drug are retained (i.e., retained in the retention fluid) and the free drug enters the filtrate.

[0062] The formulations, systems and methods provided herein include lipid-encapsulated aminoglycoside agents or lipid-associated aminoglycoside agents. The lipids used in the pharmaceutical formulations of the present invention can be synthetic, semi-synthetic or natural lipids, including phospholipids, tocopherols, sterols, fatty acids, charged lipids and cationic lipids.

[0063] In one embodiment, at least one phospholipid is present in the pharmaceutical formulation. In one embodiment, the phospholipid is phosphatidylcholine (EPC), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE), and phosphatidic acid (PA); soybean counterparts, soy phosphatidylcholine (SPC); SPG, SPS, SPI, SPE, and SPA; hydrogenated eggs and soybean counterparts (e.g., HEPC, HSPC), esters of fatty acids at the 2- and 3-positions of glycerol containing chains of 12 to 26 carbon atoms, and various head groups at the 1-position of glycerol including choline, glycerol, inositol, serine, ethanolamine, and the corresponding phosphatidic acids. The carbon chains of these fatty acids can be saturated or unsaturated, and the phospholipids can be composed of fatty acids with different chain lengths and degrees of unsaturation.

[0064] In one embodiment, the pharmaceutical formulation contains dipalmitoylphosphatidylcholine (DPPC), which is a major component of natural lung surfactant. In one embodiment, the lipid component of the pharmaceutical formulation contains DPPC and cholesterol, consists essentially of DPPC and cholesterol, or consists of DPPC and cholesterol. In a further embodiment, DPPC and cholesterol have a molar ratio in the range of about 19:1 to about 1:1, or about 9:1 to about 1:1, or about 4:1 to about 1:1, or about 2:1 to about 1:1, or about 1.86:1 to about 1:1. In yet another embodiment, DPPC and cholesterol have a molar ratio of about 2:1 or about 1:1. In one embodiment, DPPC and cholesterol are provided in an aminoglycoside formulation, for example, an aminoglycoside formulation.

[0065] Other examples of lipids used with the present invention include dimyristoylphosphatidychloline (DMPC), dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidcholine (DSPC), distearoylphosphatidylglycerol (DSPG), dioleoylphosphatidylethanolamine (DOPE), mixed phospholipids such as palmitoyl stearoyl phosphatidylcholine (PSPC), and monoacylated phospholipids such as monooleoyl - phosphatidylethanolamine (MOPE), but are not limited thereto.

[0066] In one embodiment, the at least one lipid component includes a sterol. In a further embodiment, the at least one lipid component comprises or consists essentially of or consists of a sterol and a phospholipid. Sterols used with the present invention include cholesterol, esters of cholesterol such as cholesterol hemisuccinate, salts of cholesterol such as cholesterol hydrogen sulfate and cholesterol sulfate, ergosterol, esters of ergosterol such as ergosterol hemisuccinate, salts of ergosterol such as ergosterol hydrogen sulfate and ergosterol sulfate, lanosterol, esters of lanosterol such as lanosterol hemisuccinate, salts of lanosterol such as lanosterol hydrogen sulfate, lanosterol sulfate, and tocopherol, but are not limited thereto. Tocopherol can include tocopherol, esters of tocopherol such as tocopherol hemisuccinate, salts of tocopherol such as tocopherol hydrogen sulfate and tocopherol sulfate. The term "sterol compound" encompasses sterols, tocopherols, and the like.

[0067] In one embodiment, at least one cationic lipid (positively charged lipid) is provided in the system described herein. The cationic lipids that can be used can include ammonium salts of fatty acids, phospholipids, and glycerides. Fatty acids include fatty acids having a carbon chain length of 12 to 26 carbon atoms that are saturated or unsaturated. Some specific examples include: myristylamine, palmitylamine, laurylamine, and stearylamine, dilauroyl ethylphosphocholine (DLEP), dimyristoyl ethylphosphocholine (DMEP), dipalmitoyl ethylphosphocholine (DPEP), and distearoyl ethylphosphocholine (DSEP), N-(2,3-di-(9-(Z)-octadecenyl oxy)-prop-1-yl-N,N,N-trimethylammonium chloride (DOTMA), and 1,2-bis(oleoyloxy)-3-(trimethylammonio)propane (DOTAP).

[0068] In one embodiment, at least one anionic lipid (negatively charged lipid) is provided in the system described herein. Negatively charged lipids that can be used include phosphatidyl-glycerol (PG), phosphatidic acid (PA), phosphatidylinositol (PI), and phosphatidylserine (PS), etc. Examples include DMPG, DPPG, DSPG, DMPA, DPPA, DSPA, DMPI, DPPI, DSPI, DMPS, DPPS, and DSPS.

[0069] Without wishing to be bound by theory, phosphatidylcholines such as DPPC help the uptake of aminoglycoside agents by cells in the lung (e.g., alveolar macrophages) and help maintain aminoglycoside agents in the lung. Negatively charged lipids such as PG, PA, PS, and PI are thought to play a role in reducing particle aggregation, as well as in the sustained activity characteristics of inhaled formulations and the transport (transcytosis) of formulations across the lung for systemic uptake. Without wishing to be bound by theory, sterol compounds are thought to affect the release characteristics of the formulation.

[0070] A liposome is a completely closed lipid bilayer membrane that contains a certain amount of encapsulated water. Liposomes can be unilamellar vesicles (having a single membrane bilayer) or multilamellar vesicles (onion-like structures characterized by multiple membrane bilayers each separated from the next by an aqueous layer) or combinations thereof. The bilayer is composed of two lipid monolayers having a hydrophobic "tail" region and a hydrophilic "head" region. The structure of the membrane bilayer is such that the hydrophobic (nonpolar) "tails" of the lipid monolayers are oriented towards the center of the bilayer while the hydrophilic "heads" are oriented towards the aqueous phase.

[0071] Liposomes can be prepared by various methods (see, for example, Cullis et al. (1987)). In one embodiment, one or more of the methods described in U.S. Patent Application Publication No. 2008 / 0089927 are used herein to prepare an aminoglycoside-encapsulating lipid formulation (liposome dispersion). The disclosure of U.S. Patent Application Publication No. 2008 / 0089927 is hereby incorporated by reference in its entirety for all purposes. For example, in one embodiment, at least one lipid and an aminoglycoside are mixed with a coacervate (i.e., a separate liquid phase) to form a liposome formulation. The coacervate can be formed prior to mixing with the lipid, during mixing with the lipid, or after mixing with the lipid. Also, the coacervate can be a coacervate of an active agent.

[0072] In one embodiment, a liposome dispersion is formed by dissolving one or more lipids in an organic solvent to form a lipid solution, and an aqueous solution of an aminoglycoside is mixed with the lipid solution to form an aminoglycoside coacervate. In a further embodiment, the organic solvent is ethanol. In yet another embodiment, the one or more lipids include phospholipids and sterols.

[0073] In one embodiment, liposomes are prepared by sonication, extrusion, homogenization, swelling, electroformation, inverted emulsion or reverse phase evaporation. In the Bangham method (J. Mol. Biol. (1965)), normal multilamellar vesicles (MLV) are prepared. Lenk et al. (U.S. Pat. Nos. 4,522,803, 5,030,453 and 5,169,637), Fountain et al. (U.S. Pat. No. 4,588,578) and Cullis et al. (U.S. Pat. No. 4,975,282) have disclosed methods for preparing multilamellar liposomes in which the interlamellar solute distribution in each of the aqueous compartments is substantially equal. U.S. Pat. No. 4,235,871 to Paphadjopoulos et al. discloses the preparation of oligolamellar liposomes by reverse phase evaporation. Each of these methods is suitable for use with the present invention.

[0074] Unilamellar vesicles can be prepared from MLV by several techniques, such as the extrusion techniques of U.S. Pat. Nos. 5,008,050 and 5,059,421. Sonication and homogenization can also be used to prepare small unilamellar liposomes from large liposomes (see, for example, Paphadjopoulos et al. (1968); Deamer and Uster (1983); and Chapman et al. (1968)).

[0075] In the liposome preparation of Bangham et al. (J. Mol. Biol. 13, 1965, pp. 238-252), phospholipids are suspended in an organic solvent and then evaporated to dryness, leaving a phospholipid film on the reaction vessel. Next, an appropriate amount of aqueous phase is added and the mixture is "swelled", and the resulting liposomes consisting of multilamellar vesicles (MLV) are dispersed by mechanical means. This preparation forms the basis for the development of small sonicated unilamellar vesicles and large unilamellar vesicles described by Papahadjopoulos et al. (Biochim. Biophys. Acta. 135, 1967, pp. 624-638).

[0076] Techniques for making large unilamellar vesicles (LUVs), such as reverse-phase evaporation, infusion methods, and detergent dilution, can be used to produce liposomes for use in the pharmaceutical formulations provided herein. An overview of these and other methods for making liposomes can be found in Chapter 1 of the book "Liposome", edited by Marc Ostro, Marcel Dekker, Inc., New York, 1983, which is incorporated herein by reference. Similarly, see also Szoka, Jr. et al. (Ann. Rev. Biophys. Bioeng. 9, 1980, p. 467), which is incorporated herein by reference for all purposes.

[0077] Other techniques for making liposomes include those that form reverse-phase evaporation vesicles (REV), U.S. Patent No. 4,235,871. Another type of liposome that can be used is characterized by having a substantially equal lamellar solute distribution. This type of liposome is called stable plurilamellar vesicles (SPLV) in the definition of U.S. Patent No. 4,522,803 and includes the single-phase vesicles described in U.S. Patent No. 4,588,578 and the above-mentioned freeze-thaw multilamellar vesicles (FATMLV).

[0078] Liposomes are formed using various sterols and their water-soluble derivatives, such as cholesterol hemisuccinate. See, for example, U.S. Patent No. 4,721,612. PCT Publication No. WO85 / 00968 by Mayhew et al. describes a method for reducing the toxicity of a drug by encapsulating the drug in liposomes containing α-tocopherol and certain derivatives thereof. Also, liposomes are formed using various tocopherols and their water-soluble derivatives. See PCT Publication No. 87 / 02219.

[0079] In one embodiment, the pharmaceutical preparation contains liposomes having an average diameter, as measured by the light scattering method, in the range of approximately 0.01 micron to approximately 3.0 microns, for example, about 0.2 to about 1.0 micron, before atomization. In one embodiment, the average diameter of the liposomes in the preparation is about 200 nm to about 300 nm, about 210 nm to about 290 nm, about 220 nm to about 280 nm, about 230 nm to about 280 nm, about 240 nm to about 280 nm, about 250 nm to about 280 nm, or about 260 nm to about 280 nm. The sustained activity profile of the liposome product can be controlled by the nature of the lipid membrane and by including other excipients in the composition.

[0080] In order to minimize the administration volume and reduce the patient administration time, in one embodiment, the liposomal encapsulation of aminoglycosides (for example, amikacin which is an aminoglycoside) is significantly efficient, and the L / D ratio takes a value as low as possible and / or as low as practicable, and at the same time, it is important to keep the liposomes small enough to penetrate the patient's mucus and biofilms, for example, Pseudomonas biofilms. In one embodiment, the L / D ratio in the liposomes provided herein is 0.7 or about 0.7 (w / w). In a further embodiment, the liposomes provided herein are small enough to effectively penetrate bacterial biofilms (for example, Pseudomonas biofilms). In yet another embodiment, the average diameter of the liposomes is about 260 to about 280 nm when measured by light scattering.

[0081] In one embodiment, the lipid-to-drug ratio in the pharmaceutical preparation provided herein is 3 to 1 or less, 2.5 to 1 or less, 2 to 1 or less, 1.5 to 1 or less, or 1 to 1 or less. In another embodiment, the lipid-to-drug ratio in the pharmaceutical preparation provided herein is less than 3 to 1, less than 2.5 to 1, less than 2 to 1, less than 1.5 to 1, or less than 1 to 1. In a further embodiment, the lipid-to-drug ratio is about 0.7 to 1 or less or about 0.7 to 1. In one embodiment, one of the lipids or lipid combinations in Table 1 below is used in the pharmaceutical preparation of the present invention.

Table 1

[0082] In one embodiment, the system provided herein includes an aminoglycoside formulation, such as an amikacin formulation, such as an amikacin base formulation. In one embodiment, the amount of aminoglycoside provided in the system is about 450 mg, about 500 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, about 600 mg or about 610 mg. In another embodiment, the amount of aminoglycoside provided in the system is about 500 mg to about 600 mg, or about 500 mg to about 650 mg, or about 525 mg to about 625 mg, or about 550 mg to about 600 mg. In one embodiment, the amount of aminoglycoside administered to the subject is about 560 mg and is provided in an 8 mL formulation. In one embodiment, the amount of aminoglycoside administered to the subject is about 590 mg and is provided in an 8 mL formulation. In one embodiment, the amount of aminoglycoside administered to the subject is about 600 mg and is provided in an 8 mL formulation. In one embodiment, the aminoglycoside is amikacin and the amount of amikacin provided in the system is about 450 mg, about 500 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, about 600 mg or about 610 mg. In another embodiment, the aminoglycoside is amikacin and the amount of amikacin provided in the system is about 500 mg to about 650 mg, or about 525 mg to about 625 mg, or about 550 mg to about 600 mg. In one embodiment, the aminoglycoside is amikacin and the amount of amikacin administered to the subject is about 560 mg and is provided in an 8 mL formulation. In one embodiment, the aminoglycoside is amikacin and the amount of amikacin administered to the subject is about 590 mg and is provided in an 8 mL formulation. In one embodiment, the aminoglycoside is amikacin and the amount of aminoglycoside administered to the subject is about 600 mg and is provided in an 8 mL formulation.

[0083] In one embodiment, the system provided herein includes an aminoglycoside formulation, such as amikacin (base formulation). In one embodiment, the aminoglycoside formulation provided herein contains about 60 mg / mL of aminoglycoside, about 65 mg / mL of aminoglycoside, about 70 mg / mL of aminoglycoside, about 75 mg / mL of aminoglycoside, about 80 mg / mL of aminoglycoside, about 85 mg / mL of aminoglycoside, or about 90 mg / mL of aminoglycoside. In a further embodiment, the aminoglycoside is amikacin.

[0084] In one embodiment, the system provided herein includes about 8 mL of a liposomal amikacin formulation. In one embodiment, the density of the liposomal amikacin formulation is about 1.05 grams / mL, and in one embodiment, approximately 8.4 grams of the liposomal amikacin formulation per single dose is present in the system of the present invention. In a further embodiment, the total volume of the formulation is administered to a subject in need thereof.

[0085] In one embodiment, the pharmaceutical formulation provided herein contains at least one aminoglycoside, at least one phospholipid, and a sterol. In a further embodiment, the pharmaceutical formulation contains an aminoglycoside, DPPC, and cholesterol. In one embodiment, the pharmaceutical formulation is the formulation presented in Table 2 below.

Table 2

[0086] It should be noted that increasing only the aminoglycoside concentration will not lead to a reduction in the administration time. For example, in one embodiment, as the lipid-to-drug ratio is fixed and the amikacin concentration is increased (and therefore the lipid concentration is also increased since their ratio is fixed at, for example, about 0.7:1), the viscosity of the solution also increases, which slows down the atomization time.

[0087] In one embodiment, prior to atomization of the aminoglycoside formulation, about 70% to about 100% of the aminoglycoside present in the formulation is liposome-complexed. In a further embodiment, the aminoglycoside is one aminoglycoside. In yet another embodiment, the aminoglycoside is amikacin. In another embodiment, prior to atomization, about 80% to about 99%, or about 85% to about 99%, or about 90% to about 99%, or about 95% to about 99%, or about 96% to about 99% of the aminoglycoside present in the formulation is liposome-complexed. In a further embodiment, the aminoglycoside is amikacin or tobramycin. In yet another embodiment, the aminoglycoside is amikacin. In another embodiment, prior to atomization, about 98% of the aminoglycoside present in the formulation is liposome-complexed. In a further embodiment, the aminoglycoside is amikacin or tobramycin. In yet another embodiment, the aminoglycoside is amikacin.

[0088] In one embodiment, upon atomization, about 20% to about 50% of the liposome-complexed aminoglycoside agent is released due to shear stress on the liposome. In a further embodiment, the aminoglycoside agent is amikacin. In another embodiment, upon atomization, about 25% to about 45%, or about 30% to about 40% of the liposome-complexed aminoglycoside agent is released due to shear stress on the liposome. In a further embodiment, the aminoglycoside agent is amikacin.

[0089] As presented herein, the present invention provides a method and system for treating lung infections by inhalation of a liposome aminoglycoside formulation via atomization. In one embodiment, the formulation is administered via a nebulizer that provides an aerosol mist of the formulation for delivery to the lungs of a subject.

[0090] In one embodiment, the nebulizer described herein generates (i.e., achieves the total discharge rate) an aerosol of an aminoglycoside pharmaceutical formulation at a rate greater than about 0.53 g per minute, greater than about 0.54 g per minute, greater than about 0.55 g per minute, greater than about 0.58 g per minute, greater than about 0.60 g per minute, greater than about 0.65 g per minute, or greater than about 0.70 g per minute. In another embodiment, the nebulizer described herein generates (i.e., achieves the total discharge rate) an aerosol of an aminoglycoside pharmaceutical formulation at a rate of from about 0.53 g per minute to about 0.80 g per minute, from about 0.53 g per minute to about 0.70 g per minute, from about 0.55 g per minute to about 0.70 g per minute, from about 0.53 g per minute to about 0.65 g per minute, or from about 0.60 g per minute to about 0.70 g per minute. In yet another embodiment, the nebulizer described herein generates (i.e., achieves the total discharge rate) an aerosol of an aminoglycoside pharmaceutical formulation at a rate of from about 0.53 g per minute to about 0.75 g per minute, from about 0.55 g per minute to about 0.75 g per minute, from about 0.53 g per minute to about 0.65 g per minute, or from about 0.60 g per minute to about 0.75 g per minute.

[0091] Upon atomization, the liposomes in the pharmaceutical formulation leak the drug. In one embodiment, the amount of liposome-complexed aminoglycoside after atomization is from about 45% to about 85%, or from about 50% to about 80% or from about 51% to about 77%. These percentages are also referred to herein as the "percent associated aminoglycoside after atomization". As presented herein, in one embodiment, the liposomes contain an aminoglycoside, such as amikacin. In one embodiment, the percent associated aminoglycoside after atomization is from about 60% to about 70%. In a further embodiment, the aminoglycoside is amikacin. In another embodiment, the percent associated aminoglycoside after atomization is about 67%, or from about 65% to about 70%. In a further embodiment, the aminoglycoside is amikacin.

[0092] In one embodiment, the percent of associated aminoglycoside after atomization is measured by regenerating the aerosol from air by condensation in a cold trap, and then the liquid is assayed for free aminoglycoside and encapsulated aminoglycoside (associated aminoglycoside).

[0093] In one embodiment, the MMAD of the aerosol of the pharmaceutical formulation is less than 4.9 μm, less than 4.5 μm, less than 4.3 μm, less than 4.2 μm, less than 4.1 μm, less than 4.0 μm or less than 3.5 μm when measured by ACI at a gas flow rate of about 28 L / min or with a Next Generation Impactor NGI at a gas flow rate of about 15 L / min.

[0094] In one embodiment, the MMAD of the aerosol of the pharmaceutical formulation is from about 1.0 μm to about 4.2 μm, from about 3.2 μm to about 4.2 μm, from about 3.4 μm to about 4.0 μm, from about 3.5 μm to about 4.0 μm, or from about 3.5 μm to about 4.2 μm when measured by ACI. In one embodiment, the MMAD of the aerosol of the pharmaceutical formulation is from about 2.0 μm to about 4.9 μm, from about 4.4 μm to about 4.9 μm, from about 4.5 μm to about 4.9 μm, or from about 4.6 μm to about 4.9 μm when measured by NGI.

[0095] In another embodiment, the nebulizer described herein generates an aerosol of an aminoglycoside pharmaceutical formulation at a rate greater than about 0.53 g per minute, at a rate greater than about 0.55 g per minute, or at a rate greater than about 0.60 g per minute, or at a rate of about 0.60 g per minute to about 0.70 g per minute. In a further embodiment, the FPF of the aerosol is greater than or equal to about 64% when measured by ACI, greater than or equal to about 70% when measured by ACI, greater than or equal to about 51% when measured by NGI, or greater than or equal to about 60% when measured by NGI.

[0096] In one embodiment, the system provided herein includes a nebulizer selected from an electronic mesh nebulizer, a pneumonic (jet) nebulizer, an ultrasonic nebulizer, a breath-enhanced nebulizer, and a breath-actuated nebulizer. In one embodiment, the nebulizer is portable.

[0097] The operating principle of a pneumonic nebulizer is widely known to those skilled in the art and is described, for example, in Respiratory Care, Vol. 45, No. 6, pp. 609-622 (2000). Briefly, in a pneumonic nebulizer, a pressurized gas supply is used as the driving force for liquid atomization. Compressed gas is delivered, which creates a region of negative pressure. Next, the solution to be aerosolized is delivered into the gas stream and sheared into a liquid film. This film is unstable and breaks up into droplets due to surface tension. Next, by placing a baffle in the aerosol stream, smaller particles, i.e., particles having the MMAD and FPF properties described above, can be formed. In one embodiment of a pneumonic nebulizer, the gas and the solution are mixed before interacting with the baffle away from the outlet port (nozzle). In another embodiment, the liquid and the gas are not mixed until they leave the outlet port (nozzle). In one embodiment, the gas is air, O2, and / or CO2.

[0098] In one embodiment, the droplet size and ejection rate are adjusted within a pneumonic nebulizer. However, it should be considered whether the formulation to be atomized and the nature of the formulation (e.g., associative aminoglycoside %) change with the adjustment of the nebulizer. For example, in one embodiment, the gas velocity and / or the pharmaceutical formulation velocity are adjusted to achieve the ejection rate and droplet size of the present invention. In addition to or instead of this, the flow rate of the gas and / or the solution can be adjusted to achieve the droplet size and ejection rate of the present invention. For example, an increase in the gas velocity decreased the droplet size in one embodiment. In one embodiment, the ratio of the pharmaceutical formulation flow rate to the gas flow rate is adjusted to achieve the droplet size and ejection rate of the present invention. In one embodiment, an increase in the ratio of the liquid flow rate to the gas flow rate increases the particle size.

[0099] In one embodiment, the ejection rate of the pneumonic nebulizer is increased by increasing the filling volume in the liquid reservoir. Without wishing to be bound by theory, the increase in the ejection rate may be due to a reduction in the dead volume in the nebulizer. In one embodiment, the atomization time is reduced by increasing the flow rate driving the nebulizer. See, for example, Clay et al. (1983) Lancet 2, pp. 592-594 and Hess et al. (1996) Chest 110, pp. 498-505.

[0100] In one embodiment, a reservoir bag is used to capture the aerosol during the atomization process, and then the aerosol is provided to the subject by inhalation. In another embodiment, the nebulizer provided herein includes an open vent design with a valve. In this embodiment, when the patient inhales through the nebulizer, the ejection volume of the nebulizer increases. During the exhalation phase, a one-way valve redirects the patient flow away from the nebulizer chamber.

[0101] In one embodiment, the nebulizer provided herein is a continuous nebulizer. In other words, there is no need to refill the nebulizer with the pharmaceutical formulation during a single dose administration. Conversely, the nebulizer has a volume of at least 8 mL or at least 10 mL.

[0102] In one embodiment, an aminoglycoside formulation of the present invention is delivered to a patient in need thereof using a vibrating mesh nebulizer. In one embodiment, the nebulizer membrane vibrates at an ultrasonic frequency of about 100 kHz to about 250 kHz, about 110 kHz to about 200 kHz, about 110 kHz to about 200 kHz, about 110 kHz to about 150 kHz. In one embodiment, when an electric current is applied, the nebulizer membrane vibrates at a frequency of about 117 kHz.

[0103] In one embodiment, the nebulizer provided herein does not use an air compressor and thus does not generate an air flow. In one embodiment, an aerosol is created by an aerosol head that enters the mixing chamber of the device. When the patient inhales, air enters the mixing chamber through a one-way inhalation valve on the back of the mixing chamber and carries the aerosol to the patient through the mouthpiece. During exhalation, the patient's breath flows through a one-way exhalation valve on the mouthpiece of the device. In one embodiment, the nebulizer continues to generate an aerosol in the mixing chamber, which is then inhaled by the patient during the next breath. This cycle continues until the dosing reservoir of the nebulizer is empty.

[0104] The present invention is performed, in one aspect, using one of the aerosol generators (nebulizers) illustrated in FIGS. 1, 2, 3 and 4, but is not limited thereto. Also, the system of the present invention includes, in one embodiment, a nebulizer described in European Patent Application Nos. 11169080.6 and / or 10192385.2. These applications are hereby incorporated by reference in their entirety.

[0105] Figure 1 shows a therapeutic aerosol device 1 having a membrane aerosol generator 4 with an atomization chamber 2, a mouthpiece 3, and a diaphragm 5. The diaphragm can be vibrated, for example, by an annular piezoelectric element (not shown), an example of which is described in WO1997 / 29851.

[0106] In use, a pharmaceutical formulation is placed on one side of the diaphragm 5 (see FIGS. 1, 2, and 4), and then this liquid is transported through an opening in the diaphragm 5 and released as an aerosol into the atomization chamber 2 on the other side of the diaphragm 5 (see the lower part of FIGS. 1 and 2). The patient can inhale the aerosol present in the atomization chamber 2 through the mouthpiece 3.

[0107] The diaphragm 5 includes a plurality of through-holes. When an aminoglycoside pharmaceutical formulation passes through the membrane, droplets of the aminoglycoside formulation are generated. In one embodiment, the membrane is a so-called active electronic mesh nebulizer that can vibrate, such as the eFlow® nebulizer of PARI Pharma, the HL100 nebulizer of Health and Life, or the Aeroneb Go® of Aerogen (Novartis). In a further embodiment, the membrane vibrates at an ultrasonic frequency of about 100 kHz to about 150 kHz, about 110 kHz to about 140 kHz, or about 110 kHz to about 120 kHz. In a further embodiment, the membrane vibrates at a frequency of about 117 kHz when a current is applied. In a further embodiment, the membrane is fixed and another part of the fluid reservoir or fluid supply is vibratable, a so-called passive electronic mesh nebulizer, such as the Omron MicroAir Electronic Nebulizer Model U22 or the I-Neb I-neb AAD inhalation system of Philips Respironics.

[0108] In one embodiment, the length of the nozzle portion of the through-holes formed in the membrane (e.g., a vibratable membrane) affects the total output rate (TOR) of the aerosol generator. In particular, it has been found that the length of the nozzle portion is directly proportional to the total output rate, and the shorter the nozzle portion, the higher the TOR, and vice versa.

[0109] In one embodiment, the nozzle portion is sufficiently short and has a small diameter compared to the upstream portion of the through-hole. In a further embodiment, the length of the portion upstream of the nozzle portion within the through-hole has no significant effect on the TOR.

[0110] In one embodiment, the length of the nozzle portion affects the geometric standard deviation (GSD) of the droplet size distribution of the aminoglycoside pharmaceutical formulation. A low GSD characterizes a narrow droplet size distribution (droplets of uniform size), which is advantageous for targeting aerosols to the respiratory system, for example, in the treatment of bacterial infections (such as Pseudomonas or Mycobacteria) in patients with cystic fibrosis, or in the treatment of nontuberculous mycobacteria, bronchiectasis (such as in patients with cystic fibrosis or non-cystic fibrosis), Pseudomonas or Mycobacteria in patients. That is, the longer the nozzle portion, the lower the GSD. In one embodiment, the average droplet diameter is less than 5 μm and has a GSD in the range of 1.0 to 2.2, or about 1.0 to about 2.2, or 1.5 to 2.2, or about 1.5 to about 2.2.

[0111] In one embodiment, as presented above, the system provided herein includes a nebulizer that generates an aerosol of an aminoglycoside pharmaceutical formulation at a rate greater than about 0.53 g per minute, or at a rate greater than about 0.55 g per minute. In a further embodiment, the nebulizer includes a vibratable membrane having a first side in contact with the fluid and a second side opposite the side where the droplets emerge.

[0112] A membrane, such as a stainless steel membrane, can be vibrated using a piezoelectric actuator or any other suitable means. The membrane has a plurality of through-holes penetrating the membrane in an extension direction from a first surface to a second surface. The through-holes can be formed as described above by a laser light source, electroforming, or any other suitable process. When the membrane is vibrating, an aminoglycoside pharmaceutical formulation passes through the through-holes from the first surface to the second surface, generating an aerosol on the second surface. In one embodiment, each of the through-holes includes an inlet opening and an outlet opening. In a further embodiment, each of the through-holes includes a nozzle portion extending from the outlet opening towards the inlet opening through a part of the through-hole. The nozzle portion includes the minimum diameter of the through-hole and is defined by a continuous portion of the through-hole in the extension direction bounded by a larger diameter of the through-hole. In one embodiment, the larger diameter of the through-hole is defined as 3 times, about 3 times, 2 times, about 2 times, 1.5 times, or the diameter closest to about 1.5 times the minimum diameter.

[0113] In one embodiment, the minimum diameter of the through-hole is the diameter of the outlet opening. In another embodiment, the minimum diameter of the through-hole is a diameter of about 0.5×, about 0.6×, about 0.7×, about 0.8×, or about 0.9× the diameter of the outlet opening.

[0114] In one embodiment, the nebulizer provided herein includes through-holes in which the ratio of the total length in the extension direction of at least one through-hole to the length in the extension direction of each nozzle portion of the through-holes is at least 4, or at least about 4, or at least 4.5, or at least about 4.5, or at least 5, or at least about 5, or greater than about 5. In another embodiment, the nebulizer provided herein includes through-holes in which the ratio of the total length in the extension direction of most of the through-holes to the length in the extension direction of each nozzle portion of the through-holes is at least 4, or at least about 4, or at least 4.5, or at least about 4.5, or at least 5, or at least about 5, or greater than about 5.

[0115] The extension ratios described above, in one embodiment, provide an increased total ejection rate and also provide a sufficient GSD as compared to known nebulizers. With these ratio configurations, in one embodiment, the application period is shortened, which leads to increased patient comfort and increased effectiveness of the aminoglycoside compound. This is particularly advantageous when the aminoglycoside compound in the formulation is prepared at a low concentration due to its nature and thus a larger volume of the aminoglycoside pharmaceutical formulation must be administered within an acceptable time, for example, within a single dosing session.

[0116] According to one embodiment, the nozzle portion ends in the same plane as the second surface. Therefore, the length of the nozzle portion, in one embodiment, starts from the second surface towards the first surface and is defined as the portion that reaches a diameter closest to about 3 times, about 2 times, about 2.5×, or about 1.5× the minimum diameter and is bounded there. The minimum diameter in this embodiment is the diameter of the outlet opening.

[0117] In one embodiment, the minimum diameter (i.e., one boundary of the nozzle portion) is located at the end of the nozzle portion in the extension direction adjacent to the second surface. In one embodiment, the larger diameter of the through-hole located at the other boundary of the nozzle portion is located upstream of the minimum diameter in the direction in which the fluid passes through the plurality of through-holes during operation.

[0118] According to one embodiment, the minimum diameter is less than about 4.5 μm, less than about 4.0 μm, less than about 3.5 μm, or less than about 3.0 μm.

[0119] In one embodiment, the total length of at least one through-hole in the extending direction is at least about 50 μm, at least about 60 μm, at least about 70 μm, or at least about 80 μm. In a further embodiment, the total length of at least one of the plurality of through-holes is at least about 90 μm. In one embodiment, the total length of most of the plurality of through-holes in the extending direction is at least about 50 μm, at least about 60 μm, at least about 70 μm, or at least about 80 μm. In a further embodiment, the total length of most of the plurality of through-holes is at least about 90 μm.

[0120] In one embodiment, the length of the nozzle portion is less than about 25 μm, less than about 20 μm or less than about 15 μm.

[0121] According to one embodiment, the through-hole is a through-hole formed by a laser drill and is formed in at least two steps, namely, one step for forming the nozzle portion and the remaining steps for forming the remaining portion of the through-hole.

[0122] In another embodiment, the manufacturing method used results in a substantially cylindrical or conical nozzle portion with a tolerance of less than +100% of the minimum diameter, less than +75% of the minimum diameter, less than +50% of the minimum diameter, less than +30% of the minimum diameter, less than +25% of the minimum diameter, or less than +15% of the minimum diameter.

[0123] Alternatively, or in addition, the through-hole is formed by an electroforming process. In one embodiment, the through-hole has a first funnel-shaped portion on a first surface and a second funnel-shaped portion on a second surface, and the nozzle portion is located between the first funnel-shaped portion and the second funnel-shaped portion and is defined between the outlet opening and a larger diameter. In this case, the total length of the through-hole can also be defined solely by the distance from the first surface to the outlet opening (minimum diameter).

[0124] In addition, the total output rate (TOR) can be further increased by increasing the number of through-holes provided in the membrane. In one embodiment, an increase in the number of through-holes is achieved by increasing the active perforated surface of the membrane and keeping the distance between the through-holes at the same level. In another embodiment, the number of through-holes is increased by reducing the distance between the through-holes and maintaining the active area of the membrane. In addition, combinations of the above strategies can also be used.

[0125] In one embodiment, the total output rate of the nebulizer described herein is increased by increasing the density of the through-holes in the membrane. In one embodiment, the average distance between the through-holes is about 70 μm, or about 60 μm, or about 50 μm.

[0126] In one embodiment, the membrane includes about 200 to about 8,000 through-holes, about 1,000 to about 6,000 through-holes, about 2,000 to about 5,000 through-holes, or about 2,000 to about 4,000 through-holes. In one embodiment, the number of through-holes described above increases the TOR, and the TOR increases regardless of whether the nozzle parameters are realized as described above. In one embodiment, the nebulizer provided herein includes about 3,000 through-holes. In a further embodiment, the through-holes are in a hexagonal arrangement and are located, for example, approximately at the center of the membrane (e.g., a stainless steel membrane). In a further embodiment, the average distance between the through-holes is about 70 μm.

[0127] Figure 3 shows an aerosol generator (nebulizer) disclosed in WO2001 / 032246, which is incorporated herein by reference in its entirety. This aerosol generator includes a fluid reservoir 21 for containing a pharmaceutical formulation that is released in aerosol form into the mixing chamber 3 and inhaled through the mouthpiece 4 via the opening 41.

[0128] This aerosol generator includes a vibratable membrane 22 that is vibrated using a piezoelectric actuator 23. The vibratable membrane 22 has a first surface 24 facing the fluid container 21 and a second opposite surface 25 facing the mixing chamber 3. In use, the first surface 24 of the vibratable membrane 22 contacts the fluid contained in the fluid container 21. A plurality of through holes 26 penetrating the membrane from the first surface 24 to the second surface 25 are provided in the membrane 22. In use, the fluid passes from the fluid container 21 through the through holes 26 from the first surface 24 to the second surface 25, and at this time, the membrane 22 is vibrated to generate an aerosol at the second surface 25 and discharge it into the mixing chamber 3. This aerosol can then be inhaled by the patient through the mouthpiece 4 and its inhalation opening 41 from the mixing chamber 3.

[0129] Figure 5 shows a computed tomography cross-sectional scan showing three through holes 26 of such a vibratable membrane 22. The through holes 26 of this particular embodiment are formed by laser drilling using three-stage, respectively different process parameters. In the first stage, a portion 30 is formed. In the second stage, a portion 31 is formed, and in the third stage, a nozzle portion 32 is formed. In this particular embodiment, the length of the nozzle portion 32 is about 26 μm, while the portion 31 has a length of about 51 μm. The first portion 30 has a length of about 24.5 μm. As a result, the total length of each through hole is the sum of the lengths of the portion 30, the portion 31, and the nozzle portion 32, which is about 101.5 μm in this particular embodiment. Therefore, the ratio of the total length in the extension direction E of each through hole 26 to the length in the extension direction E of the nozzle portion 32 of each is approximately 3.9.

[0130] In the embodiment of Figure 6, the first portion 30 has a length of about 27 μm, the portion 31 has a length of about 55 μm, and the nozzle portion has a length of about 19 μm. As a result, the total length of the through hole 26 is about 101 μm. Therefore, in this embodiment, the ratio of the total length of the through hole 26 to the length of the corresponding nozzle portion 32 is approximately 5.3.

[0131] The vibratable membranes of FIGS. 5 and 6 were both manufactured to have 6,000 through-holes 26. The following table (Table 3) shows the mass median diameter (MMD) determined by laser diffraction of the particles released from the second surface of the membrane, the time required to completely release a certain amount of liquid (atomization time), and the TOR. The tests were conducted using a liposomal formulation of amikacin.

Table 3

[0132] Table 3 shows that the membrane 2 with a short nozzle portion gives an increased TOR and an atomization time reduced by about 36% and 5.3 minutes shorter compared to membrane 1. Table 3 also shows that the MMD did not vary significantly for each membrane tested. This is in contrast to the differences in TOR observed for each membrane. Thus, in one embodiment, in the nebulizer described herein, the atomization time is significantly reduced compared to prior art nebulizers without affecting the droplet diameter measured by MMD.

[0133] In addition to the membranes shown in FIGS. 5 and 6, membranes having a further reduced nozzle portion and 3,000 through-holes 26 were also manufactured (membranes 3 and 4 in Table 3). Specifically, in membrane 3, laser drilling was performed to shorten the nozzle portion, and membrane 4 was manufactured to have a shorter nozzle portion than membrane 3. Table 3 shows that even with 3,000 holes (membranes 3 and 4), reducing the length of the nozzle portion increases the TOR compared to membrane 1 having 6,000 holes. Comparing membranes 3 and 4 with membrane 2 further shows that reducing the number of holes (6,000 compared to 3,000) and the length of the nozzle portion increases the TOR of the nebulizer.

[0134] In one embodiment, it is advantageous to use a laser drill process for manufacturing the through-holes rather than electroforming. The through-holes shown in FIGS. 5 and 6 manufactured by laser drilling are substantially cylindrical or conical as compared to the funnel-shaped inlet and outlet of the through-holes by electroforming as disclosed in, for example, WO01 / 18280. When the through-holes are substantially cylindrical or conical as compared to the funnel-shaped inlet and outlet of the through-holes by electroforming, the vibration of the membrane, i.e., its vibration velocity, can be transmitted to the pharmaceutical formulation over a larger area by utilizing friction. Next, the pharmaceutical formulation is ejected from the outlet opening of the through-hole due to its own inertia, resulting in a liquid jet that disintegrates to form an aerosol. Without wishing to be bound by theory, it is believed that the surface or area for transmitting energy from the membrane to the liquid is reduced because the membrane by electroforming includes a significantly bent through-hole surface.

[0135] However, the present invention can also be realized with a membrane by electroforming, in which case the nozzle portion is defined by a continuous portion of the through-hole that starts from the minimum diameter of the through-hole in the extending direction and continues towards the first surface until it reaches a diameter that is 2× or 3× the minimum diameter of the hole. In one embodiment, the total length of the through-hole is measured from the minimum diameter to the first surface.

[0136] Referring again to FIG. 1, the mouthpiece 3 has an opening 6 sealed by an elastic valve element 7 (exhalation valve) so that the patient does not need to remove the treatment device after inhaling the aerosol nor lower it from the mouth. When the patient exhales into the mouthpiece 3 and thus into the atomization chamber 2, the elastic valve element 7 opens so that the exhaled air can escape from the interior of the therapeutic aerosol. During inhalation, ambient air flows through the atomization chamber 2. The atomization chamber 2 has an opening (not shown) sealed by a further elastic valve element (inhalation valve). When the patient inhales through the mouthpiece 3 and draws in from the atomization chamber 2, the elastic valve element opens so that ambient air enters the atomization chamber and is mixed with the aerosol and remains inside the atomization chamber 2 to be inhaled. A further description of this process is presented in U.S. Patent No. 6,962,151, which is hereby incorporated by reference in its entirety for all purposes.

[0137] The nebulizer shown in FIG. 2 includes a cylindrical reservoir 10 for supplying the liquid introduced into the membrane 5. As shown in FIG. 2, the vibrating membrane 5 can be arranged on the end wall 12 of the cylindrical liquid reservoir 10 to ensure that the liquid poured into the liquid reservoir when the aerosol generator is held in the position shown in FIG. 1 comes into direct contact with the membrane 5. However, other methods can also be used to introduce the liquid into the vibrating membrane without any need to modify the design of the device of the present invention for creating a negative pressure in the liquid reservoir.

[0138] On the face facing the end wall 12, the cylindrical liquid container 10 is open. The opening is used to pour the liquid into the liquid reservoir 10. There is a protrusion 15 slightly below the opening on the outer surface 13 of the peripheral wall 14, which serves as a support when the liquid container is inserted into a suitably realized opening of the housing 35.

[0139] The open end of the liquid container 10 is closed by a flexible sealing element 16. The sealing element 16 is at the end of the peripheral wall 14 of the liquid container 10 and extends into the liquid container 10 in a pot shape, so that a conically extending wall section 17 is formed in the sealing element 16 and is closed by the flat wall section 18 of the sealing element 16. As further discussed below, since the force acts through this flat wall section 18 on the sealing element 16, in one embodiment, the flat wall section 18 is thicker than other sections of the sealing element 16. There is a distance to the conical wall section 17 at the outer peripheral part of the flat wall section 18 so that the conical wall section 17 can be folded when the flat wall section 18 moves upward compared to the display in FIG. 2.

[0140] On the surface of the flat wall section 18 on the side opposite to the inside of the liquid container, there is a protrusion including a frustoconical section 19 and a cylindrical section 20. Since the flexible material of the sealing element 16 allows the deformation of the frustoconical section 19, with this design, the protrusion can be introduced into the opening adapted so that the cylindrical section fits and can be latched.

[0141] In one embodiment, the aerosol generator 4 includes a slidable sleeve 21 having an opening of this type, which is a hollow cylinder substantially open on one side. The opening for attaching the sealing element 16 is realized in the end wall of the slidable sleeve 21. When the frustoconical section 19 is latched in a predetermined position, the end wall of the slidable sleeve 21 including the opening is on the flat sealing element wall section 18. By latching the frustoconical section 19 to the slidable sleeve, it becomes possible to transmit a force from the slidable sleeve 21 to the flat wall section 18 of the sealing element 16 so that the sealing section 18 follows the movement of the slidable sleeve 21 in the central longitudinal axis direction of the liquid container 10.

[0142] In a generalized form, the slidable sleeve 21 can be regarded as a slidable element that can also be realized as a slidable rod that can be fitted into or inserted into, for example, a drill hole. The feature of the slidable element 21 is the fact that it can be used to apply a substantially linear force on the flat wall element 18 of the sealing element 16. Overall, a decisive factor for the mode of operation of the aerosol generator of the present invention is the fact that the slidable element transmits a linear movement to the sealing element so that an increase in volume occurs within the liquid reservoir 10. Since the liquid reservoir 10 is airtight in other respects, this causes the generation of a negative pressure in the liquid reservoir 10.

[0143] The sealing element 16 and the slidable element 21 can be made integrally, that is, in one step, but from different materials. The manufacturing technology for this can be utilized such that the integrated components of the nebulizer are produced, for example, in fully automated manufacturing steps.

[0144] In one embodiment, the slidable sleeve 21 is open at the end facing the drill hole for the frustum of a cone, and at least two diametrically opposed protrusions 22 and 23 project radially inside the slidable sleeve 21. The flange 24 surrounding the slidable sleeve extends radially outward. The flange 24 is used as a support for the slidable sleeve 21 in the position shown in FIG. 5, but the protrusions 22 and 23 protruding inside the slidable sleeve 21 are used to absorb the forces acting on the slidable sleeve 21, particularly the forces parallel to the central longitudinal axis. In one embodiment, these forces are generated using two helical grooves 25 on the outer side of the peripheral wall of the rotating sleeve 26.

[0145] In one embodiment, a nebulizer can be realized using one of the protrusions 22 or 23 and one groove 25. In a further embodiment, two or more protrusions arranged to be evenly distributed and a corresponding number of grooves are provided.

[0146] In one embodiment, the rotating sleeve 26 is also a cylinder with one open side, and by having its open end disposed within the slidable sleeve 21, it faces the frustum 19 and allows the frustum 19 to enter into the rotating sleeve 26. Additionally, the rotating sleeve 26 is disposed within the slidable sleeve 21 such that the protrusions 22 and 23 are within the helical groove 25. The inclination of the helical groove 25 is designed such that when the rotating sleeve 26 is rotated relative to the slidable sleeve 21, the protrusions 22 and 23 slide along the helical groove 25, and a force in a direction parallel to the central longitudinal axis is exerted on the sliding protrusions 22 and 23, and thus on the slidable sleeve 21. This force displaces the slidable sleeve 21 in the direction of the central longitudinal axis, so that the sealing element 16, which is latched within the drill hole of the slidable sleeve using the frustum, also substantially displaces in a direction parallel to the central longitudinal axis.

[0147] The displacement of the sealing element 16 in the central longitudinal axis direction of the liquid container 10 creates a negative pressure in the liquid container 10, which depends in particular on the distance by which the slidable sleeve 21 has displaced in the direction of the central longitudinal axis. This displacement increases the initial volume V RI of the airtight liquid container 10 to a volume V RN thereby creating a negative pressure. Also, this displacement is defined by the design of the helical groove 25 in the rotating sleeve 26. Thus, the aerosol generator of the present invention ensures that a negative pressure can be created in the liquid reservoir 10 using simple structural means.

[0148] To ensure that the force applied to create a negative pressure when handling the device is kept low, the rotating sleeve 26 is realized integrally with a handle 27 having a size selected such that a user can easily rotate the handle 27, and thus the rotating sleeve 26, by hand. The handle 27 has a shape of a flat cylinder or frustum with one open side, such that a peripheral gripping region 28 where the user's hand touches to turn the handle 27 is formed at the outer edge of the handle 27.

[0149] Due to the design of the helical groove 25 and the fact that the distance the slidable sleeve 21 should move longitudinally to generate sufficient negative pressure is relatively short overall, in one embodiment, it is sufficient to rotate the handle 27, and thus the rotating sleeve 26, by a relatively small rotation angle. In one embodiment, the rotation angle is in the range of 45 degrees to 360 degrees. This embodiment enables easy handling of the device of the present invention and the therapeutic aerosol generator equipped with it.

[0150] In order to create a unit that can be operated simply and uniformly from the slidable sleeve 21 and the rotating sleeve 26 including the handle 27, in one embodiment, the aerosol generator described herein has a bearing sleeve 29 for receiving the slidable sleeve 21, which substantially includes a flat cylinder with one side open. The diameter of the peripheral wall 30 of the bearing sleeve 29 is smaller than the inner diameter of the handle 27 and, in the example of the described embodiment, is provided concentrically with the gripping region 28 of the handle 27, but is matched to the inner diameter of the cylindrical latch retaining ring 31 having a smaller diameter on the surface of the handle 27 where the rotating sleeve 26 is also arranged. On the surface of the cylindrical latch retaining ring 31 facing the rotating sleeve, an edge latch retaining edge 32 is realized that can engage with a latch retaining projection 33 spaced apart on the peripheral wall 30 of the bearing sleeve 29. This enables the handle 27 to be placed on the bearing sleeve 29, whereby, as shown in FIG. 5, the handle 27 is placed on the open end of the bearing sleeve 29 and the latch retaining edge 32 is interlaced with the latch retaining projection 33.

[0151] To hold the slidable sleeve 21, an opening is provided at the center of the sealed end of the bearing sleeve 29 so that it can be seen in FIG. 2, and the slidable sleeve 21 is disposed therein. The flange 24 of the slidable sleeve 21 is at the position shown in FIG. 2 on the surface of the end wall of the bearing sleeve 29 facing the handle. Extending into the bearing opening are two diametrically opposed protrusions 51 and 52, which project into two longitudinal grooves 53 and 54 on the edge surface of the slidable sleeve 21. The longitudinal grooves 53 and 54 run parallel to the longitudinal axis of the slidable sleeve 21. The guide protrusions 51 and 52 and the longitudinal grooves 53 and 54 provide anti-rotation locking for the slidable sleeve 21, so that the rotational movement of the rotating sleeve 26 results in a linear displacement rather than a rotation of the slidable sleeve 21. As is apparent from FIG. 2, this ensures that the slidable sleeve 21 is axially displaceable but locked against rotation and held within the combination of the handle 27 and the bearing sleeve 29. Rotating the handle 27 relative to the bearing sleeve 29 causes the rotating sleeve 26 to also rotate relative to the slidable sleeve 21, thereby moving the sliding protrusions 22 and 23 along the helical groove 25. This will axially displace the slidable sleeve 21 at the opening of the bearing sleeve 29.

[0152] It is also possible to dispense with the guide protrusions 51 and 52 in the bearing opening and the longitudinal grooves 53 and 54 in the slidable sleeve 21. In one embodiment, the guide protrusions 51 and 52 and the longitudinal grooves 53 and 54 are absent in the aerosol generator, and a large-area support for the slidable sleeve 21 that holds the frustum 19, the cylindrical section 20 of the sealing element 16, and the frustum on the flat sealing element section 18 achieves anti-rotation locking of the slidable sleeve 21 using friction. In a further embodiment, the sealing element 16 is fixed so that it cannot rotate relative to the bearing sleeve 29.

[0153] In one embodiment, an annular first sealing lip 34 concentric with the opening for holding the slidable sleeve is provided on the surface of the sealed end of the bearing sleeve 19 opposite to the handle. The diameter of the first sealing lip 34 coincides with the diameter of the peripheral wall 14 of the liquid container 10. As shown in FIG. 2, this ensures that the first sealing lip 34 presses the sealing element 16 against the liquid reservoir 10 at the end of the peripheral wall in such a way that the liquid reservoir 10 is sealed. In addition, the first sealing lip 34 can also fix the sealing element 16 so that it cannot rotate relative to the liquid reservoir 10 and the bearing sleeve 29. In one embodiment, it is not necessary to apply extreme forces to ensure that the aforementioned components of the device cannot rotate relative to each other.

[0154] In one embodiment, the required force is generated, at least to some extent, using the interaction between the handle 27 and the housing 35. In the housing, a pharmaceutical preparation reservoir is integrally realized or the pharmaceutical preparation (liquid) reservoir 10 is inserted as shown in FIG. 2. In this case, the pharmaceutical preparation reservoir 10 inserted into the casing using the edge protrusion 15 is placed at intervals on a support 36 in the housing 35 that extends radially inside the housing 35. This makes it possible to easily remove the liquid reservoir 10 from the housing 35 for cleaning. In the embodiment shown in FIG. 2, since the supports are only provided intermittently, openings for the ambient air when the patient inhales are provided, as will be described in detail below.

[0155] In FIG. 2, a rotational lock realized using the handle 27 on the one hand and the housing 35 on the other hand can be verified. Locking protrusions 62 and 63 on the housing 35 are shown. However, as long as the device of the present invention is related to the generation of negative pressure in the liquid reservoir 10, there are no special requirements regarding the design of the rotational lock.

[0156] In one embodiment, when a 8 mL volume of liquid (e.g., an aminoglycoside pharmaceutical formulation) to be released, for example, in the form of an aerosol is contained (filled or injected) in the liquid reservoir 10, the liquid reservoir 10 has a volume V of at least 16 mL, at least about 16 mL, at least 18 mL, at least about 18 mL, at least 20 mL or at least about 20 mL so as to obtain an air cushion of 8 mL or about 8 mL. RN That is, it is configured to have a volume V L of the liquid in the liquid reservoir 10 with respect to the volume V RN of at least 2.0, and the ratio between the volume V A of the gas and the volume V L of the liquid is at least 1.0. Liquid reservoirs having volumes V of about 15.5 mL, about 19.5 mL and about 22.5 mL have been shown to be efficient, and the efficiency increases with an increase in V RN . RN

[0157] In one embodiment, the ratio between V RN and V L is at least 2.0, at least about 2.0, at least 2.4, at least about 2.4, at least 2.8 or at least about 2.8. In one embodiment, the ratio between V A and V L is at least 1.0, at least 1.2, at least 1.4, at least 1.6 or at least 1.8. In another embodiment, the ratio between V A and V L is at least about 1.0, at least about 1.2, at least about 1.4, at least about 1.6 or at least about 1.8.

[0158] In one embodiment, the volume of the air cushion is at least 2 mL, at least about 2 mL, at least 4 mL, at least about 4 mL, at least 6 mL, at least about 6 mL, at least 8 mL, at least about 8 mL, at least 10 mL, at least about 10 mL, at least 11 mL, at least about 11 mL, at least 12 mL, at least about 12 mL, at least 13 mL, at least about 13 mL, at least 14 mL or at least about 14 mL. In one embodiment, the volume of the air cushion is at least about 11 mL or at least about 14 mL. In one embodiment, the volume of the air cushion is from about 6 mL to about 15 mL, V RN and V L The ratio between is at least about 2.0 to at least about 3.0. In a further embodiment, the ratio between V RN and V L is at least about 2.0 to at least about 2.8.

[0159] In one embodiment, the volume of the air cushion is about 2 mL, about 4 mL, about 6 mL, about 8 mL, about 10 mL, about 11 mL, about 12 mL, about 13 mL, or about 14 mL.

[0160] In one embodiment, the ratio of the volume V L to the volume V RN of the liquid is at least 2.0. Theoretically, the infinite expansion of the volume V RN when the liquid reservoir 10 increases will result in a nearly stable negative pressure range. In one embodiment, the ratio of the volume V L to the volume V RN of the liquid is in the range of 2.0 to 4.0, and in a further embodiment, it is 2.4 to 3.2. For different initial liquid volumes V L from 4 mL to 8 mL, two examples of the ratio range (V RN / V L ) are presented in Table 4 below.

Table 4

[0161] The systems provided herein can be used for the treatment of various pulmonary infections in subjects in need of treatment for pulmonary infections. Pulmonary infections that can be treated by the methods of the present invention (such as those in patients with cystic fibrosis) include gram-negative infections. In one embodiment, infections caused by the following bacteria are treatable with the systems and formulations provided herein: Pseudomonas (e.g., P. aeruginosa (green pus bacillus), P. paucimobilis, P. putida, P. fluorescens, and P. acidovorans), Burkholderia (e.g., B. pseudomallei, B. cepacia, B. cepacia complex, B. dolosa, B. fungorum, B. gladioli, B. multivorans, B. vietnamiensis, B. pseudomallei, B. ambifaria, B. andropogonis, B. anthina, B. brasilensis, B. caledonica, B. caribensis, B. caryophylli), Staphylococcus (e.g., S. aureus (Staphylococcus aureus), S. auricularis, S. carnosus, S. epidermidis, S. lugdunensis), methicillin-resistant Staphylococcus aureus (Staphylococcus aureus) (MRSA), Streptococcus (e.g., Streptococcus pneumoniae (pneumococcus)), Escherichia coli (E. coli), Klebsiella, Enterobacter, Serratia, Haemophilus, Yersinia pestis (plague bacillus), Mycobacterium, nontuberculous Mycobacterium (e.g., M. avium, M. avium subsp. hominissuis (MAH), M. abscessus, M. chelonae, M. bolletii, M. kansasii, M. ulcerans, M. avium, M. avium complex (MAC) (M. avium and M. intracellulare), M. conspicuum, M. kansasii, M. peregrinum, M. immunogenum, M. xenopi, M.marinum, M. malmoense, M. marinum, M. mucogenicum, M. nonchromogenicum, M. scrofulaceum, M. simiae, M. smegmatis, M. szulgai, M. terrae, M. terrae complex, M. haemophilum, M. genavense, M. asiaticum, M. shimoidei, M. gordonae, M. nonchromogenicum, M. triplex, M. lentiflavum, M. celatum, M. fortuitum, M. fortuitum complex (M. fortuitum and M. chelonae)).

[0162] In one embodiment, the system described herein is used for the treatment of infectious diseases caused by nontuberculous mycobacterial infections. In one embodiment, the system described herein is used for the treatment of infectious diseases caused by Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium or M. avium complex. In a further embodiment, a patient having cystic fibrosis is treated with one or more of the systems described herein for Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium, or Mycobacterium avium complex infections. In yet another embodiment, the Mycobacterium avium infection is Mycobacterium avium subsp. hominissuis.

[0163] In one embodiment, a patient having cystic fibrosis is treated with one of the systems provided herein for a lung infection. In a further embodiment, the lung infection is a Pseudomonas infection. In yet another embodiment, the Pseudomonas infection is P. aeruginosa. In a further embodiment, the aminoglycoside in the system is amikacin.

[0164] In one embodiment, the system provided herein is used for the treatment or prevention of Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium or Mycobacterium avium complex lung infections in patients with cystic fibrosis or non-cystic fibrosis. In a further embodiment, the system provided herein comprises a liposomal aminoglycoside formulation. In a further embodiment, the aminoglycoside is selected from amikacin, apramycin, albekacin, astromicin, capreomycin, dibekacin, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamycin or combinations thereof. In yet another embodiment, the aminoglycoside is amikacin, such as amikacin sulfate.

[0165] For the treatment of infectious diseases such as Pseudomonas aeruginosa, a major cause of chronic diseases in patients with cystic fibrosis, an obstacle is drug penetration into the sputum / biofilm barrier on epithelial cells (Figure 7). In Figure 7, the donut shape represents liposome / complexed aminoglycoside, the "+" symbol represents free aminoglycoside, the "-" symbol represents mucin, alginate, and DNA, and the solid bar symbol represents Pseudomonas aeruginosa. This barrier contains both colonized and planktonic P. aeruginosa embedded in alginate or extracellular polysaccharide derived from bacteria, each having a net negative charge, as well as DNA derived from damaged white blood cells and mucin derived from lung epithelial cells. The negative charge binds positively charged drugs such as aminoglycosides, preventing their penetration and rendering them biologically ineffective (Mendelman et al., 1985). Although not wishing to be bound by theory, encapsulation of aminoglycosides into liposomes or lipid complexes shields or partially shields the aminoglycosides from non-specific binding to the sputum / biofilm, allowing penetration of the liposome or lipid complex (containing the encapsulated aminoglycoside) (Figure 7).

[0166] In another embodiment, a patient is treated for a non-tuberculous mycobacterial lung infection with one of the systems provided herein. In a further embodiment, the system provided herein comprises a liposomal amikacin formulation.

[0167] In another embodiment, the system provided herein is used for the treatment or prevention of one or more bacterial infections in patients with cystic fibrosis. In a further embodiment, the system provided herein comprises a liposomal aminoglycoside formulation. In a further embodiment, the aminoglycoside is amikacin.

[0168] In another embodiment, the system provided herein is used for the treatment or prevention of one or more bacterial infections in patients with bronchiectasis. In a further embodiment, the system provided herein comprises a liposomal aminoglycoside formulation. In a further embodiment, the aminoglycoside is amikacin or amikacin sulfate.

[0169] In yet another embodiment, the system provided herein is used for the treatment or prevention of Pseudomonas aeruginosa lung infections in non-CF bronchiectasis patients. In a further embodiment, the system provided herein comprises a liposomal aminoglycoside formulation. In a further embodiment, the aminoglycoside is amikacin.

[0170] As presented herein, the present invention provides an aminoglycoside formulation for administration by inhalation. In one embodiment, the MMAD of the aerosol is from about 3.2 μm to about 4.2 μm when measured by an Andersen Cascade Impactor (ACI), or from about 4.4 μm to about 4.9 μm when measured by a Next Generation Impactor (NGI).

[0171] In one embodiment, the atomization time of the effective amount of the aminoglycoside formulation provided herein is less than 20 minutes, less than 18 minutes, less than 16 minutes or less than 15 minutes. In one embodiment, the atomization time of the effective amount of the aminoglycoside formulation provided herein is less than 15 minutes or less than 13 minutes. In one embodiment, the atomization time of the effective amount of the aminoglycoside formulation provided herein is about 13 minutes.

[0172] In one embodiment, the formulation described herein is administered once daily to a patient in need thereof.

Examples

[0173] The present invention is further illustrated with reference to the following examples. However, it should be noted that these examples are illustrative as in the above embodiments and should in no way be construed as limiting the scope of the present invention. Example 1: Comparison of Nebulizer Reservoir Volumes

[0174] In this example, the aerosol generator was a modified investigational eFlow® nebulizer from Pari Pharma GmbH (Germany) for use with the liposomal aminoglycoside formulation provided herein. The first aerosol generator had an initial reservoir volume V RI of 13 mL (A), the second was 17 mL (B), the third was 22 mL (C), and the fourth was 20 mL (D). That is, the first increased volume V RN was 15.5 mL, the second was 19.5 mL, the third was 24.5 mL, and the fourth was 22.5 mL.

[0175] 8 mL of the liposomal amikacin formulation was poured into reservoir 10. As shown in FIG. 8, an 8 mL air cushion provided an aerosol generation period of 14 - 16 minutes upon complete discharge of the 8 mL formulation in the reservoir. However, a 12 mL air cushion decreased the aerosol generation time to the range of 12 - approximately 13 minutes. A 17 mL air cushion further decreased the aerosol generation time to an amount in the range of 10 - 12 minutes (FIG. 6).

[0176] Furthermore, the first (A) and third (C) aerosol generators were used with 8 mL of the liposomal amikacin formulation. An initial negative pressure of 50 mbar or less was generated within the reservoir. In addition, the negative pressure during aerosol generation was measured. It is shown in FIG. 9 against the aerosol generation time. In other words, FIG. 9 shows experimental data comparing the negative pressure ranges during aerosol generation for a reservoir (C) having a volume V RN of 24.5 mL and a reservoir (A) having a volume V RN of 15.5 mL. The initial amount V L of the amikacin formulation was 8 mL and the initial negative pressure was approximately 50 mbar. The graph shows that the larger air cushion prevents the negative pressure from increasing above the critical value of 300 mbar.

[0177] The dependence of the aerosol generator efficiency (proportional to the liquid ejection rate or total ejection rate) on different negative pressures was measured with the nebulizer described above. A liposomal amikacin formulation (thixotropic) having a viscosity in the range of 5.5 - 14.5 mPa·s at a shear force of 1.1 - 7.4 Pa was used in the experiment. As shown in Figure 10, the efficiency is optimal in the negative pressure range of 150 mbar to 300 mbar. Also as shown in Figure 10, the efficiency decreases at negative pressures of approximately less than 150 mbar and above 300 mbar.

[0178] Furthermore, the same liposomal amikacin formulation as in Figure 8 was used in four different aerosol generators based on modified eFlow®. Here, the first aerosol generator (A) is a modified eFlow® with an increasing volume V RN of 19.5 mL and filled with 8 mL of the liposomal amikacin formulation.

[0179] The second aerosol generator (B) has a reservoir with an increasing volume V RN of 16 mL and filled with 8 mL of the above liposomal amikacin formulation, and the third aerosol generator (C) has an increasing volume V RN of 24.5 mL and is filled with 8 mL of the above liquid. The fourth aerosol generator has an increasing volume V RN of 22.5 mL and is filled with 8 mL of the above liposomal amikacin formulation.

[0180] Figure 11 shows the experimental data of these four aerosol generators filled with 8 mL of the liposomal amikacin formulation. These results show the aerosol generation time for completely discharging the liposomal amikacin formulation in the liquid reservoir in relation to the ratio of the increasing volume of the liquid reservoir (V L ) to the initial volume of the liquid in the liquid reservoir before use (V RN ). Figure 11 shows that for the modified aerosol generator device (A), an aerosol generation time of approximately 16 minutes was required, while the aerosol generation time was the ratio V RN / V LIt shows that it decreased with the increase. This data also shows that in the third aerosol generator device (C), the aerosol generation time could be shortened by approximately 4 minutes to less than 12 minutes.

[0181] Therefore, the data obtained in Example 1 shows that increasing the air cushion can enable the aerosol generator to operate for a long time within an efficient negative pressure range, thus significantly reducing the total aerosol generation time. Therefore, even a large amount of liquid, for example 8 mL, can be atomized (released in the form of an aerosol) within a period of less than 12 minutes. Example 2: Aerosol characteristics of amikacin formulation

[0182] Eleven liposomal amikacin formulations with different lots were examined using a modified eFlow (registered trademark) nebulizer having a modified 40-mesh membrane fabricated as described herein, a liquid volume of 8 mL, and a reservoir with the above-described air cushion (i.e., modified for use with the liposomal aminoglycoside formulations described herein). Cascade impaction was performed using either an Andersen Cascade Impactor (ACI) or a Next Generation Impactor (NGI) to determine the aerosol characteristics, namely the aerodynamic mass median diameter (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF). Measurement of aerodynamic mass median diameter (MMAD) by ACI

[0183] The Andersen Cascade Impactor (ACI) was used for MMAD measurement, and the atomization operation was performed inside a ClimateZone chamber (Westech Instruments Inc., Georgia) to maintain the temperature and relative humidity % during atomization. ClimateZone was preset to a temperature of 18 °C and a relative humidity of 50%. The ACI was assembled and loaded inside the ClimateZone. A probe thermometer (VWR dual thermometer) was attached to the surface of stage 3 of the ACI to monitor the temperature of the ACI. Atomization was started when the temperature of the ACI reached 18 ± 0.5 °C.

[0184] When the 8 mL handset was loaded with 8 mL, it was found that the ACI could not handle the entire 8 mL dose. That is, the amikacin liposome formulation deposited on the ACI plate 3 overflowed. As long as there was no liquid overflow in the ACI stage 3, it was determined that the percentage of drug distribution at each ACI stage was not affected by the amount of liposomal amikacin formulation collected inside the ACI (unpublished data). Therefore, for atomization, the nebulizer was filled with 4 mL of liposomal amikacin formulation and atomized until empty, or filled with 8 mL of liposomal amikacin formulation and atomized over a collection time of about 6 minutes (i.e., about 4 mL).

[0185] The atomized product was collected in the ACI cooled to 18 °C at a flow rate of 28.3 L / min. The atomization time was recorded, and the atomization rate was calculated based on dividing the weight difference (atomization amount) by the time interval.

[0186] After collecting the atomized product, the ACI collection plates 0, 1, 2, 3, 4, 5, 6, and 7 were taken out and placed on their respective Petri dishes. To dissolve the formulation deposited on each plate, an appropriate amount of extraction solution (20 mL for plates 2, 3, and 4; 10 mL for plates 0, 1, 5, 6, and 7) was added to each Petri dish. Samples from plates 0, 1, 2, 3, 4, 5, and 6 were further appropriately diluted with mobile phase C for HPLC analysis. The sample from plate 7 was analyzed directly by HPLC without further dilution. The ACI filter was also transferred to a 20 mL vial, 10 mL of extraction solution was added, and the capped vial was vortexed to dissolve all the formulation adhering thereto. The liquid sample from the vial was filtered (0.2 μm) into an HPLC vial for HPLC analysis. The introduction port and connectors were also rinsed with 10 mL of extraction solution to dissolve the formulation deposited thereon, the samples were collected, and HPLC analysis was performed with a two-fold dilution. Based on the amount of amikacin deposited at each stage of the impactor, the aerodynamic mass median diameter (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF) were calculated.

[0187] For the nebulizer loaded with 8 mL and atomized for 6 minutes, in order to compare the fine particle dose (FPD) across all experiments, the FPD was standardized against the volume of the atomized formulation. The FPD (standardized against the volume of the atomized formulation) was calculated according to the following equation.

Number

[0188] The next generation impactor (NGI) was also used for MMAD measurement, and the atomization operation was carried out inside a ClimateZone chamber (Westech Instruments Inc., Georgia) to maintain the temperature and RH% during atomization. ClimateZone was preset to a temperature of 18 °C and a relative humidity of 50%. The NGI was assembled and loaded inside the ClimateZone. A probe thermometer (VWR dual thermometer) was attached to the surface of the NGI to monitor the temperature of the NGI. Atomization was started when the temperature of the NGI reached 18 ± 0.5 °C.

[0189] 8 mL of the liposomal amikacin formulation was added to the nebulizer and atomized. The timer was stopped when no new aerosol was observed. The atomized product was collected at a flow rate of 15 L / min in the NGI cooled to 18 °C. The atomization time was recorded, and the atomization rate was calculated based on the weight difference (atomization amount) divided by the time interval.

[0190] After aerosol collection, the NGI tray was removed from the NGI together with the tray holder. An appropriate amount of extraction solution was added to NGI cups 1, 2, 3, 4, 5, 6, 7, and MOC to dissolve the formulation deposited in these cups. This material was transferred to volumetric flasks respectively. For NGI cups 1, 2, and 6, 25 mL volumetric flasks were used, and for NIG cups 2, 3, 4, 50 mL volumetric flasks were used. Additional extraction solution was added to the cups and transferred to the volumetric flasks again. This procedure was repeated several times to completely transfer the formulation deposited in the NGI cups to the volumetric flasks. The volumetric flasks were topped up to a final volume of 25 mL or 50 mL, shaken well, and then sampled. Samples from cups 1, 2, 3, 4, 5, 6, and 7 were further appropriately diluted with mobile phase C for HPLC analysis. Samples from MOC were analyzed directly by HPLC without further dilution. The NGI filter was also transferred to a 20 mL vial, 10 mL of extraction solution was added, and the capped vial was vortexed to dissolve all the formulation adhering thereto. The liquid sample from the vial was filtered (0.2 micron) into an HPLC vial for HPLC analysis. The inlet port and connectors were also rinsed with 10 mL of extraction solution to dissolve the formulation deposited thereon, the samples were collected, and HPLC analysis was performed with an 11-fold dilution.

[0191] Based on the amount of amikacin deposited at each stage of the impactor, MMAD, GSD, and FPF were calculated.

[0192] To compare FPD cross-sectionally for all experiments, FPD was normalized with respect to the volume of the atomized formulation. FPD (normalized with respect to the volume of the atomized formulation) was calculated according to the following equation.

Equation

[0193] The results of these experiments are presented in FIGS. 12 and 13 and Table 5 below.

Table 5-1

Table 5-2

[0194] The atomization rate study (grams of formulation atomized per minute) was conducted in a biosafety cabinet (Model 1168, Type B2, FORMA Scientific). The assembled nebulizer (handset with mouthpiece and aerosol head) was first weighed empty (W1), then a fixed volume of formulation was added and the nebulizer device was weighed again (W2). The nebulizer and timer were started, and the atomized formulation was collected in a cooling impinger at a flow rate of approximately 8 L / min (see Figure 14 for details of the experimental setup). The timer was stopped when no more aerosol was observed. The nebulizer was weighed again (W3), and the atomization time (t) was recorded. The total amount of atomized formulation was calculated as W2 - W3, and the total remaining drug amount after atomization was calculated as W3 - W1. The atomization rate of the formulation was calculated according to the following formula:

Equation

[0195] The atomization rate (in g / min) and other related results for liposomal amikacin atomized using nebulizers fabricated according to this specification (24 aerosol heads were selected and used in these studies) are recorded in Table 6.

Table 6-1

Table 6-2

Table 6-3

[0196] The free amikacin and liposome-complexed amikacin in the atomized product of Example 3 were measured. As described in Example 3, the atomized product was collected in a cooling impinger (Figure 14) at a flow rate of 8 L / min.

[0197] The atomized product collected in the impinger was rinsed with 1.5% NaCl and transferred to a 100 mL or 50 mL volumetric flask. Next, the impinger was rinsed several times with 1.5% NaCl in order to transfer all of the formulation deposited on the impinger to the flask. To measure the free amikacin concentration of the atomized product, 0.5 mL of the diluted atomized product inside the volumetric flask was taken and placed on an Amicon (registered trademark) Ultra-0.5 mL 30K centrifugal filtration device (regenerated cellulose, 30K MWCO, Millipore), and this device was centrifuged at 5000 G and 15 °C for 15 minutes. An appropriate amount of the filtrate was taken and diluted 51-fold with mobile phase C solution. The amikacin concentration was determined by HPLC. To measure the total amikacin concentration of the atomized product, an appropriate amount of the diluted atomized product inside the volumetric flask was taken, diluted 101-fold (and dissolved) in an extraction solution (perfluoropentanoic acid: 1-propanol: water (25:225:250, v / v / v)), and the amikacin concentration was determined by HPLC.

[0198] The percentage of associated amikacin after atomization was calculated by the following formula.

Equation

[0199] The percentage of associated amikacin after atomization and the total dose recovery rate from the atomization experiments described in Table 6 are summarized in Table 7. The corresponding atomization rates are also included in Table 7.

Table 7-1

Table 7-2

Table 7-3

[0200] During this study, the total concentration of amikacin in the liposomal amikacin formulation was measured using the same HPLC and amikacin standard with the remaining samples. The value obtained was 64 mg / mL amikacin. The percentage values of associated amikacin after atomization ranged from 58.1% to 72.7%, and the average value was 65.5 ± 2.6%. When 8 mL of the liposomal amikacin formulation was atomized, the total recovered amount of amikacin ranged from 426 mg to 519 mg, and the average value was 476 ± 17 mg. The calculated amount of atomized amikacin (based on the weight of the atomized liposomal amikacin formulation in Table 7) ranged from 471 mg to 501 mg, and the average value was 490 ± 8 mg. The total amikacin recovery rate ranged from 91% to 104%, and the average value was 97 ± 3% (n = 72). Liposome size

[0201] The liposomal amikacin formulation (64 mg / mL amikacin) before or after atomization was appropriately diluted with 1.5% NaCl, and the liposome particle size was measured by light scattering using a Nicomp 380 Submicron Particle Sizer (Nicomp, Santa Barbara, CA).

[0202] The liposome size of the atomized liposomal amikacin formulation was measured using 24 nebulizer aerosol heads with an 8 mL reservoir handset. The liposome size ranged from 248.9 nm to 288.6 nm, and the average was 264.8 ± 6.7 nm (n = 72). These results are presented in Table 8. The average liposome diameter before atomization was approximately 285 nm (284.5 nm ± 6.3 nm).

Table 8-1

Table 8-2

Table 8-3

[0203] All documents, patents, patent applications, publications, product descriptions, and protocols cited in this application are hereby incorporated by reference in their entirety for all purposes into this specification.

[0204] The embodiments illustrated and described in this specification are merely intended to teach those skilled in the art the best method known to the inventors for making and using the present invention. As will be understood by those skilled in the art in light of the foregoing teachings, modifications and changes can be made to the above embodiments of the present invention without departing from the present invention. Therefore, it goes without saying that the present invention can be practiced in a manner different from the specifically described methods within the scope of the claims of this application and their equivalents.

Claims

**Claim 1**: A system for treating or preventing pulmonary infections, the system comprising: (a) a pharmaceutical formulation comprising liposome-complexed amikacin, the pharmaceutical formulation being an aqueous dispersion, and the lipid component of the liposome comprising dipalmitoylphosphatidylcholine (DPPC) and cholesterol; (b) a vibrating mesh nebulizer comprising a vibratable membrane having a plurality of through-holes, each of the plurality of through-holes having an inlet opening, an outlet opening, and a nozzle portion extending from the outlet opening toward the inlet opening with a length of less than 25 μm; wherein: the ratio of the length in the extending direction of each nozzle portion of the through-holes to the total length in the extending direction of one or more of the plurality of through-holes is at least 4; the average distance between the through-holes is about 50 μm, about 60 μm, or about 70 μm; the vibrating mesh nebulizer generates an aerosol of the pharmaceutical formulation at a rate of 0.60 g per minute to about 0.80 g per minute; the fine particle fraction (FPF) of the aerosol is higher than about 64% or equal to about 64% as measured by an Andersen Cascade Impactor (ACI), or higher than about 51% or equal to about 51% as measured by a Next Generation Impactor (NGI); the percentage of associated amikacin after atomization is 60% to 70%, and the FPF refers to the fraction of the aerosol having a particle size of less than 5 μm. **Claim 2**: The system according to claim 1, wherein the amikacin is amikacin sulfate. **Claim 3**: The system according to claim 1 or 2, wherein the liposomes comprise unilamellar vesicles, multilamellar vesicles, or a mixture thereof. **Claim 4**: The system according to any one of claims 1 to 3, wherein the weight ratio of free amikacin to the liposome-complexed amikacin is about 1:100 to about 100:

1. **Claim 5**: The system according to any one of claims 1 to 4, wherein the weight ratio of free amikacin to the liposome-complexed amikacin is about 1:10 to about 10:

1. **Claim 6**: The system according to any one of claims 1 to 5, wherein the weight ratio of free amikacin to the liposome-complexed amikacin is about 0.3:1 to about 2:

1. **Claim 7**: The system according to any one of claims 1 to 6, wherein the volume of the pharmaceutical formulation is about 8 mL. **Claim 8**: The system according to any one of claims 1 to 7, wherein the aerosol has a MMAD of about 3.2 μm to about 4.2 μm when measured by ACI, or about 4.4 μm to about 4.9 μm when measured by NGI. **Claim 9**: The system according to any one of claims 1 to 8, wherein the aerosol has a MMAD of about 3.6 μm to about 3.9 μm when measured by ACI, or about 4.5 μm to about 4.8 μm when measured by NGI. **Claim 10**: The system according to any one of claims 1 to 9, wherein the FPF of the aerosol is about 64% to about 80% when measured by ACI, or about 51% to about 65% when measured by NGI. **Claim 11**: The system according to any one of claims 1 to 10, wherein the aerosol comprises a free amikacin in an amount effective to confer immediate bactericidal or immediate antibiotic activity against the lung infection, and a liposome-complexed amikacin in an amount effective to confer sustained bactericidal or sustained antibiotic activity against the lung infection. **Claim 12**: The system according to any one of claims 1 to 11, wherein the pharmaceutical formulation comprises about 500 mg to about 650 mg of amikacin. **Claim 13**: The system according to any one of claims 1 to 11, wherein the pharmaceutical formulation comprises about 550 mg to about 625 mg of amikacin. **Claim 14**: The system according to any one of claims 1 to 11, wherein the pharmaceutical formulation comprises about 550 mg to about 600 mg of amikacin. **Claim 15**: The system according to any one of claims 1 to 11, wherein the pharmaceutical formulation comprises about 560 mg of amikacin. **Claim 16**: The system according to any one of claims 1 to 11, wherein the pharmaceutical formulation comprises about 580 mg of amikacin. **Claim 17**: The system according to any one of claims 1 to 11, wherein the pharmaceutical formulation comprises about 590 mg of amikacin. **Claim 18**: The system according to any one of claims 1 to 11, wherein the pharmaceutical formulation comprises about 600 mg of amikacin. **Claim 19**: The system according to any one of claims 1 to 18, wherein the lung infection is a non-tuberculous mycobacterial lung infection. **Claim 20**: The system according to claim 19, wherein the non-tuberculous mycobacterial lung infection is an M. avium lung infection.

21. The system according to claim 20, wherein the Mycobacterium avium pulmonary infection is Mycobacterium avium subsp. hominisuis pulmonary infection.

22. The system according to claim 19, wherein the non-tuberculous mycobacterial pulmonary infection is Mycobacterium abscessus pulmonary infection.

23. The system according to claim 19, wherein the non-tuberculous mycobacterial pulmonary infection is Mycobacterium avium complex (M. avium and M. intracellulare) pulmonary infection.

24. The system according to any one of claims 1 to 23, wherein a ratio of a total length in an extending direction of one or more of the plurality of through-holes to a length in the extending direction of each nozzle portion of the through-holes is at least 4.

5.

25. The system according to any one of claims 1 to 23, wherein a ratio of a total length in an extending direction of one or more of the plurality of through-holes to a length in the extending direction of each nozzle portion of the through-holes is at least 5.

26. The system according to any one of claims 1 to 25, wherein an average distance between the through-holes is about 50 μm.

27. The system according to any one of claims 1 to 25, wherein an average distance between the through-holes is about 60 μm.

28. The system according to any one of claims 1 to 25, wherein an average distance between the through-holes is about 70 μm.

29. The system according to any one of claims 1 to 28, wherein a lipid component of the liposome consists of DPPC and cholesterol.

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