A system for treating lung infections

Abstract

We provide a system for treating lung infections. [Solution] The present invention provides a method for treating various lung infections, including mycobacterial infections, by providing a system for delivering an aerosolized liposome formulation by inhalation. In one embodiment, the present invention provides a system for treating or preventing lung infections. In one embodiment, the system comprises a pharmaceutical formulation containing 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 approximately 0.53 g per minute.

Description

[Technical Field]

[0001] Cross-references to related applications This application claims priority from U.S. Provisional Patent Application No. 61 / 649,830, filed on 21 May 2012, the entire contents of which U.S. Provisional Patent Application are incorporated herein by reference. [Background technology]

[0002] Background of the Invention Certain technologies suitable for inhalation administration utilize liposomes, where lipid complexes deliver the therapeutic effects of the drug over extended periods in the lungs. These technologies also provide sustained-activity drugs and enable targeting of the drug to the disease site, enhancing drug uptake at the site.

[0003] Liposome inhalation delivery is complicated by the fact that liposomes are susceptible to shear-induced stress during atomization, which can lead to changes in their physical characteristics (e.g., encapsulation rate, size). However, as long as these changes are reproducible and meet acceptable standards, they do not necessarily hinder drug development.

[0004] Patients with cystic fibrosis (CF) exhibit thick mucus and / or sputum secretion in the lungs, frequent secondary infections, and biofilm formation due to bacterial colonization. All of these fluids and materials pose barriers to effective targeting of infections with aminoglycosides. Liposomal aminoglycoside formulations may be helpful in combating bacterial biofilms. [Overview of the Initiative] [Means for solving the problem]

[0005] Summary of the Invention The present invention provides a system for delivering an aerosolized liposomal formulation by inhalation, and provides a method for treating various lung infections, including mycobacterial infections (e.g., lung infections caused by nontuberculous mycobacteria, also referred herein as nontuberculous mycobacterial (NTM) infections). For example, the system and method provided herein are for nontuberculous mycobacterial infections of the lung, e.g., M. avium of the lung, M. avium subsp.hominissuis (MAH), M.abscessus, M.chelonae, M.bolletii, M.kansasii, M.ulcerans, M.avium, M.avium complex (MAC) (M.avium and 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.haemophilu M. genavense, M. gordonae, M.ulcerans, M. fortuitum or M. fortuitum complex (M. fortuitum and M. chelonae) infections.

[0006] In one embodiment, the present invention provides a system for treating or preventing pulmonary infections. In one embodiment, the system comprises a pharmaceutical formulation containing 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 approximately 0.53 g per minute. In one embodiment, the mass median aerodynamic diameter (MMAD) of the aerosol is less than approximately 4.2 μm when measured by an Anderson Cascade Impactor (ACI), approximately 3.2 μm to approximately 4.2 μm when measured by an ACI, or less than approximately 4.9 μm when measured by a Next Generation Impactor (NGI), approximately 4.4 μm to approximately 4.9 μm when measured by an NGI.

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

[0008] In one embodiment, the system provided herein comprises a pharmaceutical formulation containing an aminoglycoside. In a further embodiment, the aminoglycoside is amikacin, apramycin, arbekacin, astromycin, capreomycin, dibekacin, flamycetin, gentamicin, hygromycin B, isepamycin, kanamycin, neomycin, netylmycin, paromomycin, rhodestreptomycin, ribostamycin, shisomycin, spectinomycin, streptomycin, tobramycin, verdamicin, or a combination thereof. In yet another embodiment, the aminoglycoside is amikacin. In yet another embodiment, the aminoglycoside is selected from the aminoglycosides listed in Table A below or a combination thereof. [Table A]

[0009] The pharmaceutical formulations provided herein are liposome dispersions (i.e., liposome dispersions or aqueous liposome dispersions, which may be either liposome solutions or liposome suspensions). In one embodiment, the lipid component of the liposome essentially consists 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 dipalmitoylphosphatidylcholine (DPPC) and the sterol is cholesterol.

[0010] In one embodiment, the lipid-to-drug ratio in the aminoglycoside pharmaceutical preparation (aminoglycoside liposome solution or suspension) is approximately 2:1, approximately 2:1 or less, approximately 1:1, approximately 1:1 or less, or approximately 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 is liposome-complexed prior to atomization.

[0013] In one embodiment, the liposome-complexed (also referred to herein as "liposome association") aminoglycoside percentage 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 embodiment, the present invention provides a method for treating or preventing a pulmonary infection. In one embodiment, the pulmonary infection is a pulmonary infection caused by Gram-negative bacteria (also referred to herein as a Gram-negative bacterial infection). In one embodiment, the pulmonary infection is a Pseudomonas infection, for example, a Pseudomonas aeruginosa (Pseudomonas aeruginosa) infection. In another embodiment, the pulmonary infection is caused by one of the species of the genus Pseudomonas shown in Table B below. In one embodiment, the patient receives treatment for a mycobacterial pulmonary infection with 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 embodiments described above, the patient is a patient with cystic fibrosis.

[0015] In one embodiment, a patient with cystic fibrosis receives treatment for a pulmonary infection using one of the systems provided herein. In a further embodiment, the pulmonary infection is caused by Mycobacterium abscessus, Mycobacterium avium complex, or P. aeruginosa (Pseudomonas aeruginosa). In another embodiment, the pulmonary infection is caused by 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. scrofula It is caused by nontuberculous mycobacteria selected from M. ceum, 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 embodiment, a method is provided for treating or preventing a patient's pulmonary infection. In one embodiment, the method comprises aerosolizing a pharmaceutical formulation comprising a liposome-conjugated aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion of liposomes (e.g., a liposome solution or liposome suspension) and is aerosolized at a rate exceeding approximately 0.53 grams per minute. The method further comprises administering the aerosolized pharmaceutical formulation to the patient's lungs, wherein the aerosolized pharmaceutical formulation comprises a mixture of free aminoglycosides and liposome-conjugated aminoglycosides, and the lipid component of the liposomes consists of electrically neutral lipids. In a further embodiment, the aerodynamic median mass diameter (MMAD) of the aerosol is approximately 1.0 μm to approximately 4.2 μm, as measured by ACI. In any one of the embodiments described above, the MMAD of the aerosol is approximately 3.2 μm to approximately 4.2 μm, as measured by ACI. In any of the embodiments described above, the MMAD of the aerosol, when measured by NGI, is approximately 1.0 μm to approximately 4.9 μm. In any of the embodiments described above, the MMAD of the aerosol, when measured by NGI, is approximately 4.4 μm to approximately 4.9 μm.

[0017] In one embodiment, the method comprises aerosolizing a pharmaceutical formulation containing a liposome-encompassed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate greater than approximately 0.53 grams per minute. The method further comprises administering the aerosolized pharmaceutical formulation to the lungs of a patient, wherein the aerosolized pharmaceutical formulation comprises a mixture of free aminoglycosides and liposome-encompassed aminoglycosides (e.g., aminoglycosides 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 approximately 64% when measured by ACI, or greater than or equal to approximately 51% when measured by NGI.

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

[0019] In one embodiment, the MMAD of the liposome-encompassed aminoglycoside aerosol is approximately 3.2 μm to 4.2 μm when measured by ACI, or approximately 4.4 μm to 4.9 μm when measured by NGI. In a further embodiment, the aerosol comprises an aminoglycoside and a plurality of liposomes containing DPPC and cholesterol, where approximately 65% ​​to 75% of the aminoglycoside is liposome-encompassed (e.g., encapsulated in a plurality of liposomes), and the liposome-encompassed aminoglycoside aerosol is produced at a rate exceeding approximately 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 FPF of the lipid-complexed aminoglycoside aerosol is higher than or equal to approximately 64% when measured by an Andersen Cascade Impactor (ACI), or higher than or equal to approximately 51% when measured by a Next-Generation Impactor (NGI). In a further embodiment, the aerosol comprises an aminoglycoside and a plurality of liposomes containing DPPC and cholesterol, where approximately 65% ​​to 75% of the aminoglycoside is liposome-complexed, for example, encapsulated in a plurality of liposomes, and the liposome-complexed aminoglycoside aerosol is produced at a rate exceeding approximately 0.53 grams per minute. In any of the embodiments described above, the aerosol is produced at a rate exceeding approximately 0.54 grams per minute. In any of the embodiments described above, the aerosol is produced at a rate exceeding approximately 0.55 grams per minute. In any of the embodiments described above, 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 containing DPPC and cholesterol, where approximately 65% ​​to 75% of the aminoglycoside is liposome-complexed. In a further embodiment, approximately 65% ​​to 75% of the aminoglycoside is encapsulated within the plurality of liposomes. In a further embodiment, the aerosol is produced at a rate exceeding approximately 0.53 grams per minute, a rate exceeding approximately 0.54 grams per minute, or a rate exceeding approximately 0.55 grams per minute. In a further embodiment, the aminoglycoside is amikacin (e.g., amikacin sulfate).

[0022] In one embodiment, the concentration of aminoglycoside in the liposome-conjugated aminoglycoside is approximately 50 mg / mL or higher. In a further embodiment, the concentration of aminoglycoside in the liposome-conjugated aminoglycoside is approximately 60 mg / mL or higher. In a further embodiment, the concentration of aminoglycoside in the liposome-conjugated aminoglycoside is approximately 70 mg / mL or higher, for example, approximately 70 mg / mL to approximately 75 mg / mL. In a further embodiment, the aminoglycoside is selected from the aminoglycosides shown 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 providing treatment or prevention of pulmonary infections in patients, wherein the system is (a) A pharmaceutical formulation comprising a liposome-encompassed aminoglycoside, wherein the formulation is an aqueous dispersion, and the lipid component of the liposome is an electrically neutral lipid; (b) A nebulizer that generates an aerosol of the pharmaceutical preparation at a rate exceeding approximately 0.53 g per minute. A system comprising the aerodynamic median mass diameter (MMAD) of the aerosol being less than approximately 4.2 μm when measured by an Anderson Cascade Impactor (ACI), or less than approximately 4.9 μm when measured by a Next Generation Impactor (NGI). (Item 2) A system for providing treatment or prevention of lung infections, wherein the system is (a) A pharmaceutical formulation comprising a liposome-encompassed aminoglycoside, wherein the formulation is an aqueous dispersion, and the lipid component of the liposome is an electrically neutral lipid; (b) A nebulizer that generates an aerosol of the pharmaceutical preparation at a rate exceeding approximately 0.53 g per minute. A system comprising the aerosol having a fine particle fraction (FPF) of approximately 64% or equal to approximately 64% when measured by ACI, or approximately 51% or equal to approximately 51% when measured by NGI. (Item 3) The system according to item 1 or 2, wherein the aminoglycoside is selected from amikacin, apramycin, arbekacin, astromycin, capreomycin, dibekacin, flamycetin, gentamicin, hygromycin B, isepamycin, kanamycin, neomycin, netylmycin, paromomycin, rhodestreptomycin, ribostamycin, shisomycin, spectinomycin, streptomycin, tobramycin, verdamicin, 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 comprises a unilamellar vesicle, a multilamellar vesicle, or a mixture thereof. (Item 7) The system according to any one of items 1 to 6, wherein the electrically neutral lipid comprises an electrically neutral phospholipid or an electrically neutral phospholipid and sterol. (Item 8) The system according to any one of items 1 to 7, wherein the electrically neutral lipid comprises phosphatidylcholine and sterols. (Item 9) The system according to any one of items 1 to 8, wherein the electrically neutral lipid comprises dipalmitoylphosphatidylcholine (DPPC) and sterols. (Item 10) The system according to any one of items 1 to 9, wherein the electrically neutral lipid comprises 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 comprises 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-encompassed aminoglycoside is approximately 1:100 to approximately 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-encompassed aminoglycoside is approximately 1:10 to approximately 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-encompassed aminoglycoside is approximately 0.3:1 to approximately 2:1. (Item 15) A system described in any one of items 1 to 14, wherein the volume of the pharmaceutical preparation is approximately 8 mL. (Item 16) The system according to any one of items 1 to 15, wherein the aerosol contains approximately 55% to approximately 75% liposome-encompassed amikacin. (Item 17) The system according to any one of items 1 to 16, wherein the liposome-encompassed aminoglycoside has an MMAD of approximately 3.2 μm to approximately 4.2 μm when measured by ACI, or approximately 4.4 μm to approximately 4.9 μm when measured by NGI. (Item 18) The system according to any one of items 1 to 17, wherein the liposome-encompassed aminoglycoside has an MMAD of approximately 3.6 μm to approximately 3.9 μm when measured by ACI, or approximately 4.5 μm to approximately 4.8 μm when measured by NGI. (Item 19) The system according to any one of items 1 to 18, wherein the FPF of the aerosolized formulation is greater than or equal to about 64% when measured by ACI, or greater 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 approximately 64% to approximately 80% when measured by ACI, or approximately 51% to approximately 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 preparation at a rate exceeding approximately 0.54 g per minute. (Item 22) The system according to any one of items 1 to 21, wherein the aerosol comprises a free aminoglycoside in an amount effective to provide immediate bactericidal or immediate antibiotic activity against the lung infection, and a liposome-encomplexed aminoglycoside in an amount effective to provide sustained bactericidal or sustained antibiotic activity against the lung infection. (Item 23) The system according to any one of items 1 to 22, wherein the pharmaceutical preparation contains approximately 500 mg to approximately 650 mg of aminoglycoside. (Item 24) The system according to any one of items 1 to 22, wherein the pharmaceutical preparation contains approximately 550 mg to approximately 625 mg of aminoglycoside. (Item 25) The system according to any one of items 1 to 22, wherein the aforementioned pharmaceutical preparation contains approximately 550 mg to approximately 600 mg of aminoglycoside. (Item 26) The system according to any one of items 1 to 22, wherein the pharmaceutical preparation contains approximately 560 mg of aminoglycoside. (Item 27) The system according to any one of items 1 to 22, wherein the pharmaceutical preparation contains approximately 580 mg of aminoglycoside. (Item 28) The system according to any one of items 1 to 22, wherein the pharmaceutical preparation contains approximately 590 mg of aminoglycoside. (Item 29) The system according to any one of items 1 to 22, wherein the pharmaceutical preparation contains approximately 600 mg of aminoglycoside. (Item 30) A method for treating or preventing a pulmonary infection in a patient, wherein the method is: A step of aerosolizing a pharmaceutical formulation containing a liposome-encompassed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate exceeding approximately 0.53 grams per minute; and The process of administering the aerosolized pharmaceutical preparation to the patient's lungs. Includes, The aerosolized pharmaceutical formulation comprises a mixture of free aminoglycosides and liposome-conjugated aminoglycosides. The lipid component of the liposome consists of electrically neutral lipids, and A method wherein the MMAD of the aerosol is less than approximately 4.2 μm when measured by ACI, or less than approximately 4.9 μm when measured by NGI. (Item 31) The method according to item 30, wherein the MMAD of the aerosol is approximately 3.2 μm to approximately 4.2 μm when measured by ACI, or approximately 4.4 μm to approximately 4.9 μm when measured by NGI. (Item 32) A method for providing treatment or prevention of pulmonary infection in a patient, wherein the method is: A step of aerosolizing a pharmaceutical formulation containing a liposome-encompassed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate exceeding approximately 0.53 grams per minute; and The process of administering the aerosolized pharmaceutical preparation to the patient's lungs. Includes, The aerosolized pharmaceutical formulation comprises a mixture of free aminoglycosides and liposome-conjugated aminoglycosides. The lipid component of the liposome consists of electrically neutral lipids, and A method wherein the FPF of the aerosol is greater than or equal to approximately 64% when measured by ACI, or greater than or equal to approximately 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 approximately 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 approximately 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 approximately 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 approximately 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 approximately 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 comprises approximately 70 to 75 mg / mL of amikacin, approximately 32 to 35 mg / mL of DPPC, and approximately 16 to 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 approximately 8 mL. (Item 40) The method according to any one of items 30 to 39, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astromycin, bekanamycin, voformycin, bulramycin, capreomycin, dibekacin, dactymicin, ethymicin, furamycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamycin, KA-5685, kanamycin, neomycin, netylmycin, paromomycin, prazomycin, ribostamycin, shisomycin, rhodostreptomycin, sorbistine, spectinomycin, sporalysin, streptomycin, tobramycin, verdamicin, vertilmycin, or a combination thereof. (Item 41) The method according to any one of items 30 to 40, wherein the aerosolized pharmaceutical preparation is administered once daily in one 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 or immediate antibiotic activity against the nontuberculous mycobacterial infection, and the liposome-encompassed aminoglycoside is in an amount effective to provide sustained bactericidal 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 preparation contains approximately 500 mg to approximately 650 mg of aminoglycoside. (Item 46) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains approximately 550 mg to approximately 625 mg of aminoglycoside. (Item 47) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains approximately 550 mg to approximately 600 mg of aminoglycoside. (Item 48) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains approximately 560 mg of aminoglycoside. (Item 49) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains approximately 580 mg of aminoglycoside. (Item 50) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains approximately 590 mg of aminoglycoside. (Item 51) The method according to any one of items 30 to 44, wherein the pharmaceutical preparation contains approximately 600 mg of aminoglycoside. (Item 52) A process to obtain a liposome aminoglycoside aerosol by atomizing an aqueous liposome dispersion of approximately 8 to 9 grams of aminoglycosides in less than 16 minutes, and The process of delivering the liposomal aminoglycoside aerosol to the patient's lungs by inhalation. A method for delivering liposomal aminoglycoside aerosols containing [the specified substance]. (Item 53) A step of obtaining a liposome aminoglycoside aerosol by atomizing an aqueous liposome dispersion of approximately 8 to 9 grams of aminoglycosides in approximately 10 to 15 minutes, and The process of delivering the liposomal aminoglycoside aerosol to the patient's lungs by inhalation. A method for delivering liposomal aminoglycoside aerosols containing [the specified substance]. (Item 54) The method according to item 52 or 53, wherein an aqueous liposome dispersion of aminoglycosides is atomized in less than approximately 15 minutes, less than approximately 14 minutes, less than approximately 13 minutes, less than approximately 12 minutes, or less than approximately 11 minutes. (Item 55) The method according to item 52 or 53, wherein the aqueous liposome dispersion of aminoglycosides is atomized in approximately 10 to 14 minutes, 10 to 13 minutes, 10 to 12 minutes, 10 to 11 minutes, 11 to 15 minutes, 12 to 15 minutes, 13 to 15 minutes, or 14 to 15 minutes. (Item 56) The method according to any one of items 52 to 55, wherein the liposomal aminoglycoside aerosol comprises an aminoglycoside and liposomes composed of DPPC and cholesterol, and approximately 55% to approximately 75% of the aminoglycoside is liposomally complexed. (Item 57) The liposomal aminoglycoside aerosol, When measured by ACI, the MMAD is approximately 3.2 μm to 4.2 μm, or when measured by NGI, it is approximately 4.4 μm to 4.9 μm. GSD of approximately 1.75 to 1.80; FPF that is higher than or equal to approximately 64% when measured by ACI, or higher than or equal to approximately 51% when measured by NGI; or Approximately 35 to 41 FPDs The method described in any one of items 52 to 56, wherein the method is as described in item 52 to 56. (Item 58) The method according to any one of items 52 to 57, wherein approximately 25% to approximately 35% of the liposomal aminoglycoside aerosol is deposited in the bronchi and the alveolar regions 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 the aminoglycoside and liposomes composed of DPPC and cholesterol, wherein more than 95% of the aminoglycoside is encapsulated in the liposomes. (Item 60) The method according to any one of items 52 to 59, wherein the aqueous liposomal dispersion of aminoglycosides comprises approximately 70 to approximately 75 mg / mL of aminoglycosides, approximately 32 to approximately 35 mg / mL of DPPC, and approximately 16 mg / mL to approximately 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 approximately 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 approximately 500 mg to approximately 650 mg of aminoglycoside is delivered to the patient's lungs by inhalation. (Item 65) The method according to any one of items 52 to 63, wherein approximately 550 mg to approximately 625 mg of aminoglycoside is delivered to the patient's lungs by inhalation. (Item 66) The method according to any one of items 52 to 63, wherein approximately 550 mg to approximately 600 mg of aminoglycoside is delivered to the patient's lungs by inhalation. (Item 67) The method according to any one of items 52 to 63, wherein approximately 560 mg of aminoglycoside is delivered to the patient's lungs by inhalation. (Item 68) The method according to any one of items 52 to 63, wherein approximately 580 mg of aminoglycoside is delivered to the patient's lungs by inhalation. (Item 69) The method according to any one of items 52 to 63, wherein approximately 590 mg of aminoglycoside is delivered to the patient's lungs by inhalation. (Item 70) The method according to any one of items 52 to 63, wherein approximately 600 mg of aminoglycoside is delivered to the patient's lungs by inhalation. (Item 71) A liposomal aminoglycoside aerosol comprising liposomes containing aminoglycosides, DPPC, and cholesterol, wherein approximately 65% ​​to approximately 75% of the aminoglycosides are liposome-complexed, and the liposomal aminoglycoside aerosol is produced at a rate exceeding approximately 0.53 grams per minute. (Item 72) A liposome aminoglycoside aerosol as described in item 71, produced at a rate of more than or equal to approximately 0.54 grams per minute, more than or equal to approximately 0.55 grams per minute, or more than or equal to approximately 0.60 grams per minute. (Item 73) A liposomal aminoglycoside aerosol as described in item 71 or 72, produced at a rate of approximately 0.60 to 0.70 grams per minute. (Item 74) A liposomal aminoglycoside aerosol as described in any one of items 71-73, which provides approximately 500 mg or approximately 560 mg of amikacin in a single administration session. (Item 75) A liposomal aminoglycoside aerosol as described in item 74, wherein the administration session is once daily. (Item 76) A liposome aminoglycoside aerosol according to any one of items 71 to 75, wherein the MMAD of the aerosol is less than approximately 4.2 μm when measured by ACI, or less than approximately 4.9 μm when measured by NGI. (Item 77) A liposome aminoglycoside aerosol according to any one of items 71 to 76, wherein the MMAD of the aerosol, when measured by ACI, is approximately 1.0 μm to approximately 4.2 μm, approximately 2.0 μm to approximately 4.2 μm, approximately 3.2 μm to approximately 4.2 μm, approximately 1.0 μm to approximately 4.9 μm, approximately 2.0 μm to approximately 4.9 μm, and approximately 4.4 μm to approximately 4.9 μm. (Item 78) A liposome 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 approximately 64% when measured by ACI, or higher than or equal to approximately 51% when measured by NGI. (Item 79) A liposomal aminoglycoside aerosol according to any one of items 71 to 78, wherein the aerosol contains liposome-encomplexed amikacin in a proportion greater than or equal to approximately 50%. (Item 80) A liposomal aminoglycoside aerosol according to any one of items 71 to 79, wherein the aerosol contains liposome-encomplexed amikacin in a proportion greater than or equal to approximately 60%. (Item 81) A liposomal aminoglycoside aerosol according to any one of items 71 to 80, wherein the aerosol contains approximately 55% to approximately 85% liposome-encomplexed amikacin. (Item 82) A liposomal aminoglycoside aerosol according to any one of items 71 to 81, wherein the aerosol contains approximately 55% to approximately 75% liposome-encomplexed amikacin. (Item 83) The liposome aminoglycoside aerosol according to any one of items 71 to 82, wherein the liposome comprises a unilamellar vesicle, a multilamellar vesicle, or a mixture thereof. (Item 84) The liposomal aminoglycoside aerosol described in any one of items 71 to 83, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astromycin, bekanamycin, voformycin, bruramycin, capreomycin, dibekacin, dactymicin, ethymicin, furamycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamycin, KA-5685, kanamycin, neomycin, netylmycin, paromomycin, prazomycin, ribostamycin, shisomycin, rhodostreptomycin, sorbistine, spectinomycin, sporalysin, streptomycin, tobramycin, verdamicin, vertilmycin, or a combination thereof. (Item 85) A liposomal aminoglycoside aerosol comprising a plurality of liposomes containing aminoglycosides, DPPC, and cholesterol, wherein approximately 65% ​​to approximately 75% of the aminoglycosides are liposome-complexed, and the plurality of liposomes have a diameter of approximately 245 nm to approximately 290 nm as measured by light scattering. (Item 86) A liposomal aminoglycoside aerosol as described in item 85, produced at a rate exceeding approximately 0.53 grams per minute. (Item 87) A liposomal aminoglycoside aerosol, as described in item 85 or 86, is provided with approximately 560 mg of amikacin per administration session. (Item 88) A liposomal aminoglycoside aerosol according to any one of items 85 to 87, wherein the administration session is once daily. (Item 89) A liposome aminoglycoside aerosol according to any one of items 85 to 88, wherein the MMAD of the aerosol, when measured by ACI, is approximately 1.0 μm to approximately 4.2 μm, approximately 2.0 μm to approximately 4.2 μm, approximately 3.2 μm to approximately 4.2 μm, approximately 1.0 μm to approximately 4.9 μm, approximately 2.0 μm to approximately 4.9 μm, and approximately 4.4 μm to approximately 4.9 μm. (Item 90) A liposome aminoglycoside aerosol according to any one of items 85 to 89, wherein the MMAD of the aerosol is approximately 3.2 μm to approximately 4.2 μm when measured by ACI, or approximately 4.4 μm to approximately 4.9 μm when measured by NGI. (Item 91) A liposome aminoglycoside aerosol according to any one of items 85-90, wherein the FPF of the aerosol is higher than or equal to approximately 64% when measured by ACI, or higher than or equal to approximately 51% when measured by NGI. (Item 92) The liposomal aminoglycoside aerosol according to any one of items 85 to 91, wherein the aerosol contains more than approximately 55% liposomal-conjugated amikacin, more than approximately 60% liposomal-conjugated amikacin, more than approximately 65% ​​liposomal-conjugated amikacin, or more than approximately 70% liposomal-conjugated amikacin. (Item 93) A liposomal aminoglycoside aerosol according to any one of items 85 to 92, wherein the aerosol contains approximately 55% to approximately 75% liposome-encomplexed amikacin. (Item 94) The liposome aminoglycoside aerosol according to any one of items 85 to 93, wherein the liposome comprises unilamellar vesicles, multilamellar vesicles, or a mixture thereof. (Item 95) The liposomal aminoglycoside aerosol according to any one of items 85 to 94, wherein the aminoglycoside is selected from amikacin, apramycin, arbekacin, astromycin, capreomycin, dibekacin, flamycetin, gentamicin, hygromycin B, isepamycin, kanamycin, neomycin, netylmycin, paromomycin, rhodostreptomycin, ribostamycin, shisomycin, spectinomycin, streptomycin, tobramycin, verdamicin, or a combination thereof. (Item 96) A liposomal aminoglycoside aerosol as described in any one of items 71 to 95, produced at a rate of approximately 0.55 to 0.70 grams per minute, or approximately 0.60 to 0.70 grams per minute, or approximately 0.65 to 0.70 grams per minute. (Item 97) A liposome aminoglycoside aerosol as described in any one of items 85-96, wherein the average liposome size, as measured by light scattering, is approximately 265 nm. (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 30-51 and 98, wherein the lung infection is a non-tuberculous mycobacterial infection. (Item 100) The method according to any one of items 30-51 and 98, wherein the lung infection is a Pseudomonas infection. (Item 101) The method according to any one of items 30-51 and 98, wherein the lung infection is Burkholderia infection. (Item 102) The method according to item 100, wherein the Pseudomonas infection is Pseudomonas aeruginosa infection. (Item 104) 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 described in item 99, wherein the non-tuberculous mycobacterial infection is M. avium. (Item 106) The method described in 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 a Mycobacterium avium complex (M. avium and M. intracellulare). (Item 109) The non-tuberculous mycobacterial infection is 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.sc The method described in item 99, selected from M. rofulaceum, 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. (Item 110) The system according to any one of items 1 to 29, wherein the concentration of the aminoglycoside is approximately 50 mg / mL or more, or approximately 60 mg / mL or more, or approximately 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 approximately 70 mg / mL, approximately 71 mg / mL, approximately 72 mg / mL, approximately 73 mg / mL, approximately 74 mg / mL, approximately 75 mg / mL, approximately 76 mg / mL, approximately 77 mg / mL, approximately 78 mg / mL, or approximately 79 mg / mL. (Item 112) The system according to any one of items 1 to 29, wherein the concentration of the aminoglycoside is approximately 60 mg / mL to approximately 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 30-70 and 98-109, wherein the concentration of the aminoglycoside is approximately 50 mg / mL or more, or approximately 60 mg / mL or more, or approximately 70 mg / mL or more. (Item 116) The method according to any one of items 30-70 and 98-109, wherein the concentration of the aminoglycoside is approximately 70 mg / mL, approximately 71 mg / mL, approximately 72 mg / mL, approximately 73 mg / mL, approximately 74 mg / mL, approximately 75 mg / mL, approximately 76 mg / mL, approximately 77 mg / mL, approximately 78 mg / mL, or approximately 79 mg / mL. (Item 117) The method according to any one of items 30-70 and 98-109, wherein the concentration of the aminoglycoside is approximately 60 mg / mL to approximately 80 mg / mL. (Item 118) The method according to any one of items 30-70 and 98-109, wherein the concentration of the aminoglycoside is approximately 70 mg / mL to approximately 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) A liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is approximately 50 mg / mL or more, or approximately 60 mg / mL or more, or approximately 70 mg / mL or more. (Item 122) A liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is approximately 70 mg / mL, approximately 71 mg / mL, approximately 72 mg / mL, approximately 73 mg / mL, approximately 74 mg / mL, approximately 75 mg / mL, approximately 76 mg / mL, approximately 77 mg / mL, approximately 78 mg / mL, or approximately 79 mg / mL. (Item 123) A liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is approximately 60 mg / mL to approximately 80 mg / mL. (Item 124) A liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is approximately 70 mg / mL to approximately 80 mg / mL. (Item 125) A 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) A 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 described in any one of items 1 to 29, wherein the lung infection is a Pseudomonas lung infection. (Item 129) A system according to any one of items 1 to 29, wherein the lung infection is Burkholderia lung infection. (Item 130) The system described in item 128, wherein the Pseudomonas infection is Pseudomonas aeruginosa pulmonary infection. (Item 131) The Burkholderia lung infection is caused by B.pseudomallei, B.cepacia, B.cepacia complex, B.dolosa, B.fungorum, B.gladioli, B.multivorans, B.vietnamiensis, B.pse udomallei, B. ambifaria, B. andropogonis, B. anthina, B. brasilensis, B. caledonica, B. caribensis or B. caryophylli lung infection. (Item 132) The non-tuberculous mycobacterial lung infections are caused by M.avium, M.abscessus, M.chelonae, M.bolletii, M.kansasii, M.ulcerans, M.avium, M.avium complex (MAC) (M.avium and M.intracellula). re), M.conspicuum, M.kansasii, M.peregrinum, M.immunogenum, M.xenopi, M.marinum, M.malmoense, M.marinum, M.mucogenicum, M.nonchromogenicum, M.s M. crofulaceum, 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) pulmonary infections, or combinations thereof, as described in item 127. (Item 133) The system described in item 132, wherein the non-tuberculous mycobacterial lung infection is M. abscessus lung infection. (Item 134) The system described in item 132, wherein the non-tuberculous mycobacterial lung infection is M. avium lung infection. (Item 135) The system described in item 134, wherein the non-tuberculous mycobacterial lung infection is a M. avium subsp. hominissuis lung infection. (Item 137) The method according to any one of items 30-39, 41 and 44-70, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astromycin, bekanamycin, voformycin, bruramycin, capreomycin, dibekacin, dactymicin, ethymicin, furamycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamycin, KA-5685, kanamycin, neomycin, netylmycin, paromomycin, prazomycin, ribostamycin, shisomycin, rhodostreptomycin, sorbistine, spectinomycin, sporalysin, streptomycin, tobramycin, verdamicin, vertilmycin, or a combination thereof. (Item 138) The system described in any one of items 1-3, 6-29, and 110-114, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astromycin, bekanamycin, voformycin, bruramycin, capreomycin, dibekacin, dactymicin, ethymicin, furamycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamycin, KA-5685, kanamycin, neomycin, netylmycin, paromomycin, prazomycin, ribostamycin, shisomycin, rhodostreptomycin, sorbistine, spectinomycin, sporalysin, streptomycin, tobramycin, verdamicin, vertilmycin, or a combination thereof. (Item 139) The system according to any one of items 1-29, 110-114, and 138, wherein the aqueous dispersion is an aqueous suspension of liposome-encompassed aminoglycosides. (Item 140) The system according to any one of items 1-29, 110-114, and 138, wherein the aqueous dispersion is an aqueous solution of liposome-complexed aminoglycosides. (Item 141) The method according to any one of items 30-70, 115-120, and 137, wherein the aqueous dispersion is an aqueous suspension of liposome-complexed aminoglycosides. (Item 142) The method according to any one of items 30-70, 115-120, 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 30-51 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 liposome-complexed aminoglycosides. (Item 145) The method according to item 143, wherein the aqueous dispersion is an aqueous solution of a liposome-complexed aminoglycoside. (Item 146) A system described in any one of items 1 to 29, wherein the aforementioned lung infection is associated with bronchiectasis. [Brief explanation of the drawing]

[0023] [Figure 1] Figure 1 shows a nebulizer (aerosol generator) that can realize the present invention.

[0024] [Figure 2] Figure 2 is a magnified view of the nebulizer diagram shown in Figure 1.

[0025] [Figure 3] Figure 3 is a cross-sectional view of a commonly known aerosol generator described in WO2001 / 032246.

[0026] [Figure 4]Figure 4 shows an image of the PARI eFlow® nebulizer modified for use with the aminoglycoside formulations described herein, and an enlarged view of the nebulizer membrane.

[0027] [Figure 5] Figure 5 is a computed tomography (CT) cross-sectional image showing a membrane with a relatively long nozzle portion.

[0028] [Figure 6] Figure 6 shows a computed tomography (CT) cross-sectional image of a stainless steel film with a relatively short nozzle portion.

[0029] [Figure 7] Figure 7 is a schematic cross-sectional view of sputum / biofilm, for example, seen in patients with cystic fibrosis.

[0030] [Figure 8] Figure 8 is a graph of the aerosol formation period (atomization time) at the time of complete liquid release in the reservoir, as a function of the initial gas cushion (VA) in the reservoir.

[0031] [Figure 9] Figure 9 is a graph of negative pressure in a nebulizer as a function of the time required for aerosol generation (atomization time) until the complete release of the pharmaceutical formulation from the liquid reservoir.

[0032] [Figure 10] Figure 10 is a graph of aerosol generation efficiency as a function of negative pressure in a nebulizer.

[0033] [Figure 11] Figure 11 is a graph of the time for aerosol formation (atomization time) when the liquid is completely released, as a function of the ratio (VRN / VL) between the increasing volume VRN of the reservoir and the initial volume (VL) of the liquid in the reservoir.

[0034] [Figure 12] Figure 12 is a graph showing the MMAD of aerosolized formulations as a function of the atomization rate of each formulation.

[0035] [Figure 13] Figure 13 is a graph showing the FPF of aerosolized formulations as a function of the atomization rate of each formulation.

[0036] [Figure 14] Figure 14 is a schematic diagram of the system used for aerosol recovery for post-atomization research. [Modes for carrying out the invention]

[0037] Detailed description of the invention The inventions described herein are, in part, directed to a system for administering an aminoglycoside pharmaceutical formulation to a target lung, for example, to treat lung injury.

[0038] The term “to treat” includes (1) preventing or delaying the onset of clinical symptoms of the condition, disorder, or condition in a subject who is suffering from or may be predisposed to such a condition but has not yet experienced or presented any clinical or quasi-clinical symptoms of the condition, disorder, or condition; (2) inhibiting the condition, disorder, or condition (i.e., suppressing, reducing, or delaying the onset of at least one clinical or quasi-clinical symptom of the disease, or, in the case of maintenance treatment, a relapse thereof); and / or (3) mitigating the condition (i.e., causing a reduction in at least one clinical or quasi-clinical symptom of the condition, disorder, or condition). The benefit to the subject being treated is statistically significant, or at least perceptible to the subject or to the physician.

[0039] In one embodiment, the system and formulation provided herein can treat lung infections caused by the following bacteria: Pseudomonas (e.g., P. aeruginosa, P. paucimobilis, P. putida, P. fluorescens, and P. acidovans), Burkholderia (e.g., B. pseudomallei, B. cepacia, B. cepacia complex, B. dolosa, B. fungorum, B. gladioli, B. multiv) orans, B.vietnamiensis, B.pseudomallei, B.ambifaria, B.andropogonis, B.anthina, B.brasilensis, B.caledonica, B.caribensis, B.cary ophylli), 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 mycobacterium).

[0040] In one embodiment, a patient receives treatment for a nontuberculous mycobacterial lung infection using 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 system provided herein is used to treat a patient having a pulmonary infection caused by Pseudomonas. In a further embodiment, the pulmonary infection is caused by a species of the genus Pseudomonas selected from the species shown in Table B below. [Table B-1] [Table B-2]

[0042] Nontuberculous mycobacterial lung infections include, in one embodiment, 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, and M. nonchromogenicu. The following are selected from M. m, 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, the nontuberculous mycobacterial lung infection is M. abscessus or M. avium. In a further embodiment, the M. avium infection is M. avium subsp. hominissuis. In one embodiment, the nontuberculous mycobacterial lung infection is refractory nontuberculous mycobacterial lung infection.

[0043] In another embodiment, a patient with cystic fibrosis receives treatment for a bacterial infection using one of the systems provided herein. In yet another embodiment, the bacterial infection is a pulmonary infection caused by Pseudomonas aeruginosa. In yet another embodiment, the patient receives treatment for a pulmonary infection associated with bronchiectasis using one of the systems provided herein.

[0044] As used herein, “prevention” may mean the complete prevention of an infection or disease, or the prevention of the onset of symptoms of such infection or disease; the delay of the onset of an infection or disease or symptoms of such infection or disease; or a reduction in the severity of any subsequent infection or disease or symptoms of such infection or disease.

[0045] The term "antibacterial" is recognized in the art and refers to the ability of the compounds of this 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 dose" means the amount of aminoglycoside (e.g., amikacin) used in the present invention that is sufficient to produce the desired therapeutic response. The effective dose formulations provided herein include both free aminoglycosides and liposomal-conjugated aminoglycosides. For example, liposomal-conjugated aminoglycosides include, in one embodiment, aminoglycosides encapsulated in liposomes, aminoglycosides complexed with liposomes, or a combination thereof.

[0048] In one embodiment, the aminoglycoside is selected from amikacin, apramycin, arbekacin, astromycin, capreomycin, dibekacin, furamycetin, gentamicin, hygromycin B, isepamycin, kanamycin, neomycin, netylmycin, paromomycin, rhodostreptomycin, ribostamycin, shisomycin, spectinomycin, streptomycin, tobramycin, or verdamicin. In another embodiment, the aminoglycoside is selected from the aminoglycosides listed in Table C below. [Table C]

[0049] In one embodiment, the aminoglycoside is an aminoglycoside free base, a salt thereof, a solvate, or another non-covalent derivative. In a further embodiment, the aminoglycoside is amikacin. Suitable aminoglycosides for use in the drug formulations of the present invention include pharmaceutically acceptable addition salts and complexes of the drug. Where a compound may have one or more chiral centers, unless otherwise specified, the present invention includes each of the unique racemic compounds and each of the unique non-racemic compounds. Where the activator has an unsaturated carbon-carbon double bond, both cis (Z) and trans (E) isomers are included in the scope of the present invention. Where the activator exists in tautomer forms such as keto-enol tautomers, each tautomer is considered to be included in the present invention. In one embodiment, amikacin exists in the drug formulation as an amikacin base, or an amikacin salt, for example, 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 therapeutic response can be any response that the user (e.g., clinician) would recognize as an effective response to treatment. Generally, a therapeutic response would be the reduction, inhibition, delay, or prevention of the growth or proliferation of one or more of the aforementioned bacteria, or the killing of one or more of the aforementioned bacteria. A therapeutic response may also be reflected in improvements in lung function, such as forced expiratory volume in one second (FEV1). Furthermore, determining an appropriate treatment duration, appropriate dose, and any potential concomitant treatments based on an assessment of the therapeutic response is within the scope of the skills of those skilled in the art.

[0051] "Liposome dispersion" refers to a solution or suspension containing multiple liposomes.

[0052] As used herein, "aerosol" refers to a gaseous suspension of liquid particles. The aerosols provided herein include particles of liposome dispersions.

[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 airway. 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 discharge rate.

[0054] The process of converting a bulk liquid into tiny droplets using air is called atomization. Pneumatic nebulizers require a pressurized gas supply as the driving force for liquid atomization. Ultrasonic nebulizers use electricity introduced by a piezoelectric element in the liquid reservoir to convert the liquid into breathable droplets. Various types of nebulizers are described in Respiratory Care, Vol. 45, No. 6, pp. 609-622 (2000), and their disclosures are incorporated herein by reference. 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 aerosols having a particle size less than 5 μm in diameter, as measured by cascade impaction. FPF is typically expressed as a percentage.

[0056] "Median mass diameter" or "MMD" is determined by laser diffraction or impactor measurement and is the average particle diameter based on mass.

[0057] The "aerodynamic median mass diameter" or "MMAD" is standardized for the aerodynamic separation of aqueous aerosol droplets and is determined by impactor measurements, such as the Andersen Cascade Impactor (ACI) or the Next Generation Impactor (NGI). In one embodiment, the gas flow velocity is 28 liters per minute for the Andersen Cascade Impactor (ACI) and 15 liters per minute for the Next Generation Impactor (NGI). The "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. Treatment is achieved through the delivery of an aminoglycoside formulation by inhalation via atomization. In one embodiment, the pharmaceutical formulation comprises an aminoglycoside, for example, an aminoglycoside.

[0059] The pharmaceutical formulations provided herein are liposome dispersions. More specifically, the pharmaceutical formulations are dispersions comprising "liposome-complexed aminoglycosides" or "liposome-encapsulated aminoglycosides." "Liposome-complexed aminoglycosides" encompass embodiments in which aminoglycosides (or combinations of aminoglycosides) are encapsulated in liposomes, and encompass any form of aminoglycoside composition in which at least about 1% by weight of the aminoglycosides is associated with liposomes, either as part of a complex with liposomes, or as liposomes in which the aminoglycosides may be in an aqueous or hydrophobic bilayer phase, or present in the interface head group region of the liposome bilayer.

[0060] In one embodiment, the lipid component of the liposome includes electrically neutral lipids, positively charged lipids, charged lipids, or a combination thereof. In another embodiment, the lipid component includes electrically neutral lipids. In yet another embodiment, the lipid component consists essentially of electrically neutral lipids. In yet another embodiment, the lipid component consists of electrically neutral lipids, such as sterols and phospholipids.

[0061] As presented above, the liposome-complexed aminoglycoside embodiments include embodiments in which the aminoglycoside is encapsulated in liposomes. In addition, liposome-complexed aminoglycosides represent 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 liposomes, or as liposomes in which the aminoglycoside may be in an aqueous or hydrophobic bilayer phase, or present in the interface head group region of the liposome bilayer. In one embodiment, before atomization, 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 thus associated. The association is measured in one embodiment by separation by filtration such that the lipid and the lipid-associated drug are retained (i.e., retained in a retaining solution) and the free drug enters the filtrate.

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

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

[0064] In one embodiment, the pharmaceutical formulation contains dipalmitoylphosphatidylcholine (DPPC), 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 in conjunction with the present invention include, but are not limited to, dimyristoylphosphatidychloline (DMPC), dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylcholine (DSPC), distearoylphosphatidylglycerol (DSPG), dioleylphosphatidylethanolamine (DOPE), mixed phospholipids, such as palmitoylstearoylphosphatidylcholine (PSPC), and monoacylated phospholipids, such as monooleoylphosphatidylethanolamine (MOPE).

[0066] In one embodiment, the at least one lipid component comprises a sterol. In a further embodiment, the at least one lipid component comprises a sterol and a phospholipid, or is essentially composed of a sterol and a phospholipid, or is composed of a sterol and a phospholipid. Sterols used in conjunction with the present invention include, but are not limited to, cholesterol, cholesterol esters such as cholesterol hemisuccinate, cholesterol salts such as cholesterol hydrogen sulfate and cholesterol sulfate, ergosterol, ergosterol esters such as ergosterol hemisuccinate, ergosterol salts such as ergosterol hydrogen sulfate and ergosterol sulfate, lanosterol, lanosterol esters such as lanosterol hemisuccinate, lanosterol salts such as lanosterol hydrogen sulfate and lanosterol sulfate, and tocopherol. Tocopherols may include tocopherol, tocopherol esters such as tocopherol hemisuccinate, and tocopherol salts 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 used may include ammonium salts of fatty acids, phospholipids, and glycerides. The fatty acids include saturated or unsaturated fatty acids with a carbon chain length of 12 to 26 carbon atoms. Some specific examples include myristylamine, palmitylamine, laurylamine, and stearylamine, dilauroylethyl phosphocholine (DLEP), dimyristoylethyl phosphocholine (DMEP), dipalmitoylethyl phosphocholine (DPEP), and distearoylethyl phosphocholine (DSEP), N-(2,3-di-(9-(Z)-octadecenyloxy)-propa-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 (loading lipid) is provided in the system described herein. Usable loading lipids include phosphatidyl glycerol (PG), phosphatidic acid (PA), phosphatidylinositol (PI), and phosphatidylserine (PS). Examples include DMPG, DPPG, DSPG, DMPA, DPPA, DSPA, DMPI, DPPI, DSPI, DMPS, DPPS, and DSPS.

[0069] While we do not wish to be bound by theory, phosphatidylcholines such as DPPC are thought to help with the uptake of aminoglycosides by cells in the lung (e.g., alveolar macrophages) and help maintain aminoglycosides in the lungs. Loaded electrolipids such as PG, PA, PS, and PI are thought to play a role in the sustained activity characteristics of inhaled formulations, as well as in the transport of formulations across the lungs for systemic uptake (transcytosis), in addition to reducing particle aggregation. While we do not wish to be bound by theory, sterol compounds are thought to affect the release characteristics of formulations.

[0070] Liposomes are completely closed lipid bilayer membranes containing a certain amount of encapsulated water. Liposomes can be unilamellar vesicles (having a single membrane bilayer), multilamellar vesicles (an onion-like structure characterized by multiple membrane bilayers, each separated from the next by an aqueous layer), or a combination thereof. The bilayer consists of two lipid monolayers, each having a hydrophobic "tail" region and a hydrophilic "head" region. The structure of the membrane bilayer is such that the hydrophobic (nonpolar) "tail" of the lipid monolayer is oriented toward the center of the bilayer, while the hydrophilic "head" is oriented toward the aqueous phase.

[0071] Liposomes can be prepared by various methods (see, for example, Cullis et al. (1987)). In one embodiment, one or more methods described in U.S. Patent Application Publication 2008 / 0089927 are used here to prepare an aminoglycoside-encapsulated lipid formulation (liposomal dispersion). The disclosures of U.S. Patent Application Publication 2008 / 0089927 are incorporated herein by reference in their original form 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 liposomal formulation. The coacervate may be formed prior to, during, or after mixing with the lipids. The coacervate may also be an activator coacervate.

[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 aminoglycoside coacervate is formed by mixing an aqueous solution of aminoglycosides with the lipid solution. 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. The Bangham method (J. Mol. Biol. (1965)) produces typical multilamellar vesicles (MLVs). Lenk et al. (US Patents 4,522,803, 5,030,453, and 5,169,637), Fountain et al. (US Patent 4,588,578), and Cullis et al. (US Patent 4,975,282) disclose methods for preparing multilamellar liposomes in which the interlayer solute distribution in each aqueous compartment is substantially equal. Paphadjopoulos et al. (US Patent 4,235,871) discloses the preparation of oligolamellar liposomes by reverse-phase evaporation. Each of these methods is suitable for use in conjunction with the present invention.

[0074] Unilamellar vesicles can be produced from MLVs using several techniques, for example, by the extrusion technique described in U.S. Patent No. 5,008,050 and U.S. Patent No. 5,059,421. Sonication and homogenization can also be used to produce 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 method described by 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 to "swell" the mixture, and the resulting liposomes, consisting of multilamellar vesicles (MLVs), are dispersed by mechanical means. This preparation forms the basis for the development of small ultrasonically treated unilamellar vesicles and large unilamellar vesicles described by Papahadjopoulos et al. (Biochim.Biophys.Acta.135,1967,pp.624-638).

[0076] Using techniques for producing large unilamellar vesicles (LUVs), such as reverse-phase evaporation, infusion, and detergent dilution, liposomes for use in the pharmaceutical formulations provided herein can be prepared. An overview of these and other methods for preparing 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.

[0077] Other techniques for creating liposomes include forming reverse-phase evaporation vesicles (REVs), as described in U.S. Patent No. 4,235,871. Another type of liposome that can be used is characterized by having substantially equal lamellar solute distributions. This type of liposome is called stable plurilamellar vesicles (SPLVs) as defined in U.S. Patent No. 4,522,803, and includes the single-phase vesicles described in U.S. Patent No. 4,588,578, and the freeze-thaw multilamellar vesicles (FATMLVs) described above.

[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. Mayhew et al., PCT Publication No. WO85 / 00968, describes a method for reducing drug toxicity by encapsulating a drug with liposomes containing α-tocopherol and certain derivatives. Liposomes are also formed using various tocopherols and their water-soluble derivatives. See PCT Publication No. 87 / 02219.

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

[0080] To minimize the dose volume and reduce patient administration time, in one embodiment, it is important that liposome encapsulation of the aminoglycoside (e.g., amikacin, which is an aminoglycoside) is remarkably efficient, and that the L / D ratio is as low as possible and / or practically feasible, while at the same time keeping the liposomes small enough to penetrate patient mucus and biofilms, e.g., 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 (e.g., Pseudomonas biofilms). In yet another embodiment, the average diameter of the liposomes is about 260 to about 280 nm, as measured by light scattering.

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

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

[0083] In one embodiment, the system provided herein comprises an aminoglycoside formulation, such as amikacin (a base formulation). In one embodiment, the aminoglycoside formulation provided herein comprises 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 contains approximately 8 mL of liposomal amikacin formulation. In one embodiment, the density of the liposomal amikacin formulation is approximately 1.05 g / mL, and in one embodiment, approximately 8.4 g of liposomal amikacin formulation per dose is present in the system of the present invention. In a further embodiment, the entire volume of the formulation is administered to a subject requiring it.

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

[0086] It should be noted that simply increasing the aminoglycoside concentration alone will not necessarily lead to a reduction in 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 the ratio of the two 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, approximately 70% to 100% of the aminoglycosides present in the aminoglycoside formulation are liposome-conjugated before spraying. In a further embodiment, the aminoglycoside is a single aminoglycoside. In yet another embodiment, the aminoglycoside is amikacin. In yet another embodiment, approximately 80% to 99%, or approximately 85% to 99%, or approximately 90% to 99%, or approximately 95% to 99%, or approximately 96% to 99%, of the aminoglycosides present in the formulation are liposome-conjugated before spraying. In a further embodiment, the aminoglycoside is amikacin or tobramycin. In yet another embodiment, the aminoglycoside is amikacin. In yet another embodiment, approximately 98% of the aminoglycosides present in the formulation are liposome-conjugated before spraying. In yet another embodiment, the aminoglycoside is amikacin or tobramycin. In yet another embodiment, the aminoglycoside is amikacin.

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

[0089] As presented herein, the present invention provides a method and system for treating pulmonary infections by inhalation of a liposomal 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 target lung.

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

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

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

[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 approximately 28 L / min, or by a next-generation impactor NGI at a gas flow rate of approximately 15 L / min.

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

[0095] In another embodiment, the nebulizer described herein generates an aerosol of an aminoglycoside pharmaceutical formulation at a rate greater than approximately 0.53 g per minute, greater than approximately 0.55 g per minute, greater than approximately 0.60 g per minute, or at a rate of approximately 0.60 g to approximately 0.70 g per minute. In yet another embodiment, the FPF of the aerosol is greater than or equal to approximately 64% when measured by ACI, greater than or equal to approximately 70% when measured by ACI, greater than or equal to approximately 51% when measured by NGI, or greater than or equal to approximately 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 pneumatic nebulizers 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, pneumatic nebulizers use pressurized gas supply as the driving force for liquid atomization. Compressed gas is delivered, which creates a negative pressure region. Next, the solution to be aerosolized is delivered into the gas flow and sheared into a liquid film. This film is unstable and, due to surface tension, breaks into droplets. Next, by placing a baffle in the aerosol flow, smaller particles, i.e., particles having the MMAD and FPF properties described above, can be formed. In one embodiment of the pneumatic nebulizer, the gas and solution are mixed before interacting with the baffle after leaving the outlet port (nozzle). In another embodiment, mixing does not occur until the liquid and gas leave the outlet port (nozzle). In one embodiment, the gas is air, O2 and / or CO2.

[0098] In one embodiment, the droplet size and discharge rate are adjusted within a pneumonic nebulizer. However, the formulation being atomized and whether the properties of the formulation (e.g., associated aminoglycoside %) change with nebulizer adjustment should be considered. For example, in one embodiment, the gas rate and / or pharmaceutical formulation rate are adjusted to achieve the discharge rate and droplet size of the present invention. In addition to, or instead of, the flow rate of the gas and / or solution can be adjusted to achieve the droplet size and discharge rate of the present invention. For example, increasing the gas rate, in one embodiment, reduced the droplet size. In one embodiment, the ratio of pharmaceutical formulation flow rate to gas flow rate is adjusted to achieve the droplet size and discharge rate of the present invention. In one embodiment, increasing the ratio of liquid flow rate to gas flow rate increased the droplet size.

[0099] In one embodiment, the discharge rate of a pneumonic nebulizer is increased by increasing the filling volume in the liquid reservoir. While we do not wish to be bound by theory, the increase in discharge rate is likely due to a reduction in dead volume in the nebulizer. In another 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 the aerosol is then delivered to the subject by inhalation. In another embodiment, the nebulizer provided herein includes an open vent design with a valve. In this embodiment, as the patient inhales through the nebulizer, the nebulizer discharge volume increases. During the expiratory 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. Conversely, the nebulizer has a capacity of at least 8 mL or at least 10 mL.

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

[0103] In one embodiment, the nebulizer provided herein does not use an air compressor and therefore does not generate airflow. In one embodiment, the aerosol is generated 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 located at the rear of the mixing chamber, delivering 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 aerosol in the mixing chamber, which is then inhaled by the patient during the next breath. This cycle continues until the medication reservoir of the nebulizer is empty.

[0104] In one embodiment, the present invention is carried out using, but is not limited to, one of the aerosol generators (nebulizers) illustrated in Figures 1, 2, 3, and 4. Furthermore, in one embodiment, the system of the present invention includes the nebulizer described in European Patent Application No. 11169080.6 and / or No. 10192385.2. These applications are incorporated herein by direct reference.

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

[0106] During use, the pharmaceutical formulation is placed on one side of the vibrating membrane 5 (see Figures 1, 2, and 4), and this liquid is then transported through an opening in the vibrating membrane 5 and released as an aerosol into the atomizing chamber 2 on the other side of the vibrating membrane 5 (see the bottom of Figures 1 and 2). The patient can inhale the aerosol present in the atomizing chamber 2 through the mouthpiece 3.

[0107] The vibrating membrane 5 includes a plurality of through-holes. When the aminoglycoside pharmaceutical formulation passes through the membrane, droplets of the aminoglycoside formulation are generated. In one embodiment, the membrane is vibrable, a so-called active electronic mesh nebulizer, such as PARI Pharma's eFlow® nebulizer, Health and Life's HL100 nebulizer, or Aerogen (Novartis)'s Aeroneb Go®. In a further embodiment, the membrane vibrates at ultrasonic frequencies 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 an electric current is applied. In a further embodiment, the membrane is fixed and another part of the fluid reservoir or fluid supply section is vibrable, a so-called passive electronic mesh nebulizer, such as the Omron MicroAir U22 electronic nebulizer or Philips Respironics' I-Neb I-Neb AAD inhalation system.

[0108] In one embodiment, the length of the nozzle portion of a through-hole formed in a membrane (e.g., a vibrating membrane) affects the total discharge velocity (TOR) of the aerosol generator. In particular, it has been found that the length of the nozzle portion is directly proportional to the total discharge velocity, with shorter nozzle portions resulting in higher TOR, and vice versa.

[0109] In one embodiment, the nozzle portion is sufficiently short and small in diameter compared to the upstream portion of the through hole. In a further embodiment, the length of the upstream portion of the nozzle portion within the through hole does not have a significant effect on 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 with uniform size), which is advantageous for aerosol targeting to the respiratory system, for example, for the treatment of bacterial infections (e.g., Pseudomonas or Mycobacteria) in patients with cystic fibrosis, or for the treatment of non-tuberculous mycobacteria, bronchiectasis (e.g., for the treatment of patients with cystic fibrosis or non-cystic fibrosis), Pseudomonas or Mycobacteria. In other words, the longer the nozzle portion, the lower the GSD. In one embodiment, the average droplet diameter is less than 5 μm and the GSD is 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 exceeding approximately 0.53 g per minute or at a rate exceeding approximately 0.55 g per minute. In a further embodiment, the nebulizer includes a vibrable membrane having a first surface in contact with the fluid and a second surface opposite to it from which droplets emerge.

[0112] A film, such as a stainless steel film, can be vibrated using a piezoelectric actuator or any other suitable means. The film has a plurality of through-holes that penetrate the film in an elongated 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. As the film vibrates, 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 through-hole includes an inlet opening and an outlet opening. In a further embodiment, each through-hole includes a nozzle portion extending from the outlet opening through a portion of the through-hole toward the inlet opening. The nozzle portion is defined by a continuous portion of the through-hole in the elongated direction, including the minimum diameter of the through-hole and bounded by a larger diameter of the through-hole. In one embodiment, the larger diameter of the through-hole is defined as the diameter closest to 3 times, about 3 times, 2 times, about 2 times, 1.5 times, or 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 approximately 0.5 ×, 0.6 ×, 0.7 ×, 0.8 ×, or 0.9 × the diameter of the outlet opening.

[0114] In one embodiment, the nebulizer provided herein includes through-holes such that 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-hole 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 such that the ratio of the total length in the extension direction of most through-holes to the length in the extension direction of each nozzle portion of the through-hole 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 discharge rate compared to known nebulizers, and also provide sufficient GSD. These ratio configurations, in one embodiment, result in a shorter administration period, which leads to increased patient comfort and efficacy of the aminoglycoside compound. This is particularly advantageous when the aminoglycoside compound in the formulation is prepared at low concentrations due to its properties, and therefore a larger volume of the aminoglycoside pharmaceutical formulation must be administered within an acceptable time, for example, a single dosing session.

[0116] In one embodiment, the nozzle portion ends on the same plane as the second surface. Therefore, in one embodiment, the length of the nozzle portion is defined as the portion that begins from the second surface toward the first surface and ends at a diameter closest to approximately 3 times, approximately 2 times, approximately 2.5 ×, or approximately 1.5 × the minimum diameter. In this embodiment, the minimum diameter is the diameter of the outlet opening.

[0117] In one embodiment, the smallest diameter (i.e., one boundary of the nozzle portion) is located at the end of the nozzle portion in the extensional 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 smallest diameter in the direction through which the fluid passes during operation.

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

[0119] In one embodiment, the total length in the longitudinal direction of at least one through-hole 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 multiple through-holes is at least about 90 μm. In one embodiment, the total length in the longitudinal direction of most of the multiple through-holes 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 multiple through-holes is at least about 90 μm.

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

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

[0122] In another embodiment, the manufacturing method used results in a substantially cylindrical or conical nozzle portion with tolerances 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 to the above, 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, with the nozzle portion located between the first and second funnel-shaped portions and 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 discharge rate (TOR) can be further increased by increasing the number of through-holes in the membrane. In one embodiment, the increase in the number of through-holes is achieved by increasing the active perforated surface area of ​​the membrane and keeping the distance between the through-holes at the same level. In another embodiment, the increase in the number of through-holes is achieved by reducing the distance between the through-holes and maintaining the active surface area of ​​the membrane. In addition, a combination of the above strategies can also be used.

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

[0126] In one embodiment, the film contains approximately 200 to 8,000 through-holes, approximately 1,000 to 6,000 through-holes, approximately 2,000 to 5,000 through-holes, or approximately 2,000 to 4,000 through-holes. In one embodiment, the number of through-holes increases the TOR, which increases regardless of whether the nozzle parameters are achieved as described above. In one embodiment, the nebulizer provided herein contains approximately 3,000 through-holes. In a further embodiment, the through-holes are arranged in a hexagonal pattern and are positioned approximately in the center of the film (e.g., a stainless steel film). In a further embodiment, the average distance between the through-holes is approximately 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 which is released in the form of an aerosol into a mixing chamber 3 and inhaled through an opening 41 using a mouthpiece 4.

[0128] This aerosol generator includes a vibrating membrane 22 that is vibrated using a piezoelectric actuator 23. The vibrating membrane 22 has a first surface 24 facing the fluid container 21 and a second opposite surface 25 facing the mixing chamber 3. During use, the first surface 24 of the vibrating membrane 22 is in contact with the fluid contained in the fluid container 21. Multiple through-holes 26 are provided in the membrane 22, extending from the first surface 24 to the second surface 25. During use, the fluid passes from the fluid container 21 through the through-holes 26 to the first surface 24 and then to the second surface 25, causing the membrane 22 to vibrate in order to generate an aerosol on the second surface 25 and release 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 vibratory membrane 22. In this particular embodiment, the through-holes 26 are formed by laser drilling in three stages, each with 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 approximately 26 μm, while the length of the first portion 31 is approximately 51 μm. The first portion 30 is approximately 24.5 μm in length. As a result, the total length of each through-hole is the sum of the lengths of portions 30, 31, and 32, which in this particular embodiment is approximately 101.5 μm. 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 each nozzle portion 32 is approximately 3.9.

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

[0131] Both the vibratory membranes in Figures 5 and 6 were manufactured to have 6,000 through-pores 26. The table below (Table 3) shows the median mass diameter (MMD) determined by laser diffraction, the time required to completely release a certain amount of liquid (atomization time), and the TOR of the particles emitted from the second surface of the membrane. The tests were conducted using a liposome formulation of amikacin. [Table 3]

[0132] Table 3 shows that membrane 2, with its shorter nozzle portion, yields increased TOR and a reduced atomization time of 5.3 minutes, approximately 36% less than membrane 1. Table 3 also shows that MMD did not vary significantly between the tested membranes, in contrast to the differences in TOR observed between membranes. Thus, in one embodiment, the nebulizer described herein significantly reduces atomization time compared to conventional nebulizers without affecting droplet diameter as measured by MMD.

[0133] In addition to the membranes shown in Figures 5 and 6, membranes with a further reduced nozzle portion and 3,000 through-holes 26 were also manufactured (membrane 3 and membrane 4 in Table 3). Specifically, membrane 3 was laser-drilled 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 (membrane 3 and membrane 4), reducing the length of the nozzle portion increases the TOR compared to membrane 1, which has 6,000 holes. Comparing membranes 3 and 4 with membrane 2, it is further shown that the TOR of the nebulizer increases due to the larger number of holes (6,000 compared to 3,000) and the reduced length of the nozzle portion.

[0134] In one embodiment, it is advantageous to use a laser drilling process rather than electroforming for the production of through-holes. The through-holes produced by laser drilling, as shown in Figures 5 and 6, are substantially cylindrical or conical compared to the funnel-shaped inlets and outlets of through-holes produced by electroforming, such as those disclosed in WO01 / 18280. When the through-hole is substantially cylindrical or conical compared to the funnel-shaped inlets and outlets of electroforming through-holes, the vibration of the membrane, i.e., its vibrational velocity, can be transmitted to the pharmaceutical formulation over a wider area by utilizing friction. The pharmaceutical formulation is then ejected from the exit opening of the through-hole due to its own inertia, resulting in a liquid jet that collapses to form an aerosol. While we do not wish to be bound by theory, it is thought that electroforming membranes include a significantly curved through-hole surface, thus reducing the surface or area available for energy transfer from the membrane to the liquid.

[0135] However, the present invention can also be realized with a film formed by electroforming, in which case the nozzle portion is defined by a continuous portion of through holes that extends in the longitudinal direction, starting from the minimum diameter of the through hole and continuing toward the first surface until it reaches a diameter 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 Figure 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 or take it out of their mouth after inhaling the aerosol. When the patient exhales into the mouthpiece 3 and therefore into the atomizing chamber 2, the elastic valve element 7 opens so that the exhaled air can escape from inside the therapeutic aerosol. During inhalation, ambient air flows through the atomizing chamber 2. The atomizing 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 atomizing chamber 2, the elastic valve element opens so that ambient air enters the atomizing chamber, mixes with the aerosol, and remains inside the atomizing chamber 2 to be inhaled. A further explanation of this process is presented in U.S. Patent No. 6,962,151, which is incorporated herein by reference in its original form for all purposes.

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

[0138] On the side facing the end wall 12, a cylindrical liquid container 10 is open. The opening is used to pour liquid into the liquid reservoir 10. Slightly below the opening on the outer surface 13 of the peripheral wall 14 is a projection 15, which serves as a support when the liquid container is inserted into the appropriately 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 located at the end of the peripheral wall 14 of the liquid container 10 and extends in a vase-like shape into the interior of the liquid container 10, thereby forming a cone-shaped wall section 17 on the sealing element 16, which is closed by a flat wall section 18 of the sealing element 16. As will be discussed further below, forces act through this flat wall section 18 on the sealing element 16, so in one embodiment, the flat wall section 18 is thicker than the other sections of the sealing element 16. There is a distance on the outer periphery of the flat wall section 18 to the cone wall section 17 such that the cone wall section 17 can be folded when the flat wall section 18 moves upward compared to the representation in Figure 2.

[0140] On the flat wall section 18 opposite the interior of the liquid container, there is a projection including a frustoconical section 19 and a cylindrical section 20. The flexible material of the sealing element 16 allows the frustoconical section 19 to deform, so this design allows the projection to be introduced into an opening that is fitted to accommodate the cylindrical section and then latched in place.

[0141] In one embodiment, the aerosol generator 4 includes a sliding sleeve 21 having this type of opening, which is a hollow cylinder with substantially one side open. The opening for mounting the sealing element 16 is realized in the end wall of the sliding sleeve 21. When the frustocone 19 is latched in place, the end wall of the sliding sleeve 21, including the opening, lies on the flat sealing element wall section 18. The latching of the frustocone 19 to the sliding sleeve allows force to be transmitted from the sliding sleeve 21 to the flat wall section 18 of the sealing element 16 so that the sealing section 18 follows the movement of the sliding sleeve 21 in the central longitudinal axis direction of the liquid container 10.

[0142] In a generalized form, the sliding sleeve 21 can be seen as a sliding element that can be realized, for example, as a sliding rod that can be fitted into or inserted into a drilled hole. A feature of the sliding element 21 is that it can be used to apply substantially linear forces on the flat wall element 18 of the sealing element 16. Overall, a decisive factor for the operating mode of the aerosol generator of the present invention is the fact that the sliding element transmits linear motion to the sealing element so that an increase in volume occurs in the liquid reservoir 10. Since the liquid reservoir 10 is otherwise airtight, this causes the generation of negative pressure in the liquid reservoir 10.

[0143] The sealing element 16 and the sliding element 21 can be manufactured as a single unit, i.e., in a single process, but from different materials. Manufacturing techniques for this purpose can be utilized, for example, to produce the integrated components of a nebulizer in a fully automated manufacturing step.

[0144] In one embodiment, the sliding sleeve 21 is open at an end facing a drilled hole for a frustocone, but at least two diametrically opposed projections 22 and 23 protrude radially into the interior of the sliding sleeve 21. A flange 24 surrounding the sliding sleeve extends radially outward. The flange 24 is used as a support for the sliding sleeve 21 in the position shown in Figure 5, but the projections 22 and 23 protruding into the interior of the sliding sleeve 21 are used to absorb forces acting on the sliding sleeve 21, particularly those parallel to the central longitudinal axis. In one embodiment, these forces are generated using two helical grooves 25 on the outside of the peripheral wall of the rotating sleeve 26.

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

[0146] In one embodiment, the rotating sleeve 26 is also an open cylinder with one open end positioned within the sliding sleeve 21, facing the frustocone 19, allowing the frustocone 19 to enter the rotating sleeve 26. In addition, the rotating sleeve 26 is positioned within the sliding sleeve 21 such that projections 22 and 23 are located 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 sliding sleeve 21, the projections 22 and 23 slide along the helical groove 25, and a force parallel to the central longitudinal axis is exerted on the sliding projections 22 and 23, and therefore on the sliding sleeve 21. This force displaces the sliding sleeve 21 in the direction of the central longitudinal axis, so that the sealing element 16, which is latched into the drilled hole of the sliding sleeve using the frustocone, is also substantially displaced in a direction parallel to the central longitudinal axis.

[0147] The displacement of the sealing element 16 in the direction of the central longitudinal axis of the liquid container 10 generates a negative pressure in the liquid container 10, which is determined in particular by the distance the sliding sleeve 21 is displaced in the direction of the central longitudinal axis. This displacement affects the initial volume V of the airtight liquid container 10. RI Volume V RN By increasing this, negative pressure is generated. Furthermore, 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 negative pressure in the liquid reservoir 10 can be generated in the relevant region using simple structural means.

[0148] To ensure that the force applied to generate negative pressure when handling the device is kept low, the rotating sleeve 26 is realized in conjunction with a handle 27 having a size selected so that the user can rotate the handle 27, and therefore the rotating sleeve 26, by hand without difficulty. The handle 27 has substantially the shape of a flat cylinder or frustocone with one open face such that a marginal gripping area 28, which the user's hand touches to turn the handle 27, is formed on the outer edge of the handle 27.

[0149] Due to the design of the helical groove 25 and the relatively short overall distance over which the sliding sleeve 21 must travel longitudinally to generate sufficient negative pressure, in one embodiment, it is sufficient to rotate the handle 27, and therefore the rotating sleeve 26, by a relatively small angle. In one embodiment, the angle of rotation is in the range of 45 to 360 degrees. This embodiment allows for easy handling of the apparatus of the present invention and the therapeutic aerosol generator equipped therewith.

[0150] To create a unit that can be operated easily and uniformly from a sliding sleeve 21 and a rotating sleeve 26 including a handle 27, in one embodiment the aerosol generator described herein has a bearing sleeve 29 for receiving the sliding sleeve 21, which substantially comprises a flat cylinder with one open surface. The diameter of the circumferential wall 30 of the bearing sleeve 29 is smaller than the inner diameter of the handle 27 and, in the example of the embodiment described herein, is provided concentrically with the gripping area 28 of the handle 27, but the inner diameter of a cylindrical latching ring 31 having a smaller diameter is also matched on the surface of the handle 27 on which the rotating sleeve 26 is also located. On the surface of the cylindrical latching ring 31 facing the rotating sleeve, a marginal latching edge 32 is provided which can engage with a spaced-out latching projection 33 on the circumferential wall 30 of the bearing sleeve 29. This allows the handle 27 to be placed on the bearing sleeve 29, so that, as shown in Figure 5, the handle 27 is placed on the open end of the bearing sleeve 29 and the latching edge 32 interlatches with the latching projection 33.

[0151] To hold the sliding sleeve 21, an opening is provided at the center of the sealing end of the bearing sleeve 29, as can be seen in Figure 2, and the sliding sleeve 21 is positioned within it. The flange 24 of the sliding sleeve 21 is located on the surface of the end wall of the bearing sleeve 29 facing the handle, as shown in Figure 2. Extending into the bearing opening are two diametrically opposed projections 51 and 52, which protrude into two longitudinal grooves 53 and 54 on the peripheral surface of the sliding sleeve 21. The longitudinal grooves 53 and 54 run parallel to the longitudinal axis of the sliding sleeve 21. The guide projections 51 and 52 and the longitudinal grooves 53 and 54 act as anti-rotation locking for the sliding sleeve 21, so that the rotational motion of the rotating sleeve 26 results in a linear displacement of the sliding sleeve 21 rather than rotation. As is clear from Figure 2, this ensures that the sliding sleeve 21 is axially displaceable but locked to rotation, and is held in place during the combination of the handle 27 and the bearing sleeve 29. When the handle 27 is rotated relative to the bearing sleeve 29, the rotating sleeve 26 also rotates relative to the sliding sleeve 21, thereby causing the sliding projections 22 and 23 to move along the helical groove 25. This causes the sliding sleeve 21 to be axially displaced at the opening of the bearing sleeve 29.

[0152] It is also possible to omit the guide projections 51 and 52 in the bearing opening and the longitudinal grooves 53 and 54 in the sliding sleeve 21. In one embodiment, the guide projections 51 and 52 and the longitudinal grooves 53 and 54 are absent in the aerosol generator, and a large-area support for the sliding sleeve 21 that holds the frustocone 19, the cylindrical section 20 of the sealing element 16, and the frustocone on the flat sealing element section 18 uses friction to achieve anti-rotation locking of the sliding sleeve 21. In a further embodiment, the sealing element 16 is fixed so as not to rotate relative to the bearing sleeve 29.

[0153] In one embodiment, an annular first sealing lip 34 is provided on the sealing end of the bearing sleeve 19, on the surface opposite the handle, concentric with the opening that holds the sliding sleeve. The diameter of the first sealing lip 34 matches the diameter of the peripheral wall 14 of the liquid container 10. As shown in Figure 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 force 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, and the housing either integrally incorporates a pharmaceutical product reservoir or a pharmaceutical product (liquid) reservoir 10 is inserted as shown in Figure 2. In this case, the pharmaceutical product reservoir 10, inserted into the casing using the marginal projections 15, is spaced apart on support structures 36 within the housing 35 that extend radially into the housing 35. This allows the liquid reservoir 10 to be easily removed from the housing 35 for cleaning. In the embodiment shown in Figure 2, since the support structures are only spaced apart, openings are provided for ambient air when the patient inhales, as will be detailed below.

[0155] Figure 2 shows the rotation lock, which is implemented using the handle 27 on one side and the housing 35 on the other. Locking protrusions 62 and 63 on the housing 35 are shown. However, as long as the apparatus of the present invention is related to generating negative pressure in the liquid reservoir 10, there are no special requirements regarding the design of the rotation lock.

[0156] In one embodiment, when a liquid (e.g., an aminoglycoside pharmaceutical preparation) with a volume of 8 mL to be released in the form of an aerosol, for example, 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 for the initial volume V of the liquid in the liquid reservoir 10 RN such that the ratio is at least 2.0, and the ratio between the volume V of the gas A and the volume V of the liquid L 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, V RN and V L The ratio between 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 is at least 2.0. Theoretically, the infinite expansion of the volume V RN during the increase of the liquid reservoir 10 would result in a nearly stable negative pressure range. In one embodiment, the ratio of the volume V L to the volume V RN 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 of 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 system provided herein can be used to treat various lung infections in subjects requiring treatment for lung infections. Lung infections that can be treated by the method of the present invention (e.g., in patients with cystic fibrosis) include Gram-negative infections. In one embodiment, infections caused by the following bacteria can be treated with the systems and formulations provided herein: Pseudomonas (e.g., P. aeruginosa, P. paucimobilis, P. putida, P. fluorescens, and P. acidovans), Burkholderia (e.g., B. pseudomallei, B. cepacia, B. cepacia complex, B. dolosa, B. fungorum, B. gladioli, B. multivorans, B. vietnamiensis, B. pseudomallei, B. amphifaria, 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), and methicillin-resistant Staphylococcus. aureus (MRSA), Streptococcus (e.g., Streptococcus pneumoniae), Escherichia coli, Klebsiella, Enterobacter, Serratia, Haemophilus, Yersinia pestis (plague bacterium), Mycobacterium, non-tuberculous 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.gen avense, 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 to treat an infection caused by a non-tuberculous mycobacterial infection. In another embodiment, the system described herein is used to treat an infection caused by Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium, or M. avium complex. In a further embodiment, a patient with cystic fibrosis is treated with one or more of the systems described herein for an infection of Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium, or Mycobacterium avium complex. In yet another embodiment, the Mycobacterium avium infection is Mycobacterium avium subsp. hominissuis.

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

[0164] In one embodiment, the system provided herein is used to treat or prevent 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, arbekacin, astromycin, capreomycin, dibekacin, furamycetin, gentamicin, hygromycin B, isepamycin, kanamycin, neomycin, netylmycin, paromomycin, rhodostreptomycin, ribostamycin, shisomycin, spectinomycin, streptomycin, tobramycin, verdamicin, or a combination thereof. In yet another embodiment, the aminoglycoside is amikacin, for example, amikacin sulfate.

[0165] A major obstacle to treating infectious diseases such as Pseudomonas aeruginosa, a major cause of chronic disease in patients with cystic fibrosis, is drug penetration into the sputum / biofilm barrier on epithelial cells (Figure 7). In Figure 7, donut shapes represent liposomes / complexed aminoglycosides, "+" symbols represent free aminoglycosides, "-" symbols represent mucin, arginate, and DNA, and solid bar symbols represent Pseudomonas aeruginosa. This barrier includes both colonized and planktonic P. aeruginosa embedded in bacterial arginate or extracellular polysaccharides, as well as DNA from damaged leukocytes and mucin from lung epithelial cells, all of which have a net negative charge. Negative charges bind positively charged drugs such as aminoglycosides, preventing their penetration and rendering them biologically ineffective (Mendelman et al., 1985). While we do not wish to be bound by theory, encapsulation of aminoglycosides in liposomes or lipid complexes shields or partially shields them from nonspecific binding to sputum / biofilms, allowing the penetration of liposomes or lipid complexes (containing the encapsulated aminoglycosides) (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 yet another embodiment, the system provided herein comprises a liposomal amikacin formulation.

[0167] In another embodiment, the system provided herein is used to treat or prevent 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 to treat or prevent 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 to treat or prevent Pseudomonas aeruginosa (Pseudomonas aeruginosa) lung infections in patients with non-CF bronchiectasis. In yet another embodiment, the system provided herein comprises a liposomal aminoglycoside formulation. In yet another embodiment, the aminoglycoside is amikacin.

[0170] As presented herein, the present invention provides an aminoglycoside formulation administered by inhalation. In one embodiment, the MMAD of the aerosol is approximately 3.2 μm to approximately 4.2 μm when measured by an Andersen cascade impactor (ACI), or approximately 4.4 μm to approximately 4.9 μm when measured by a next-generation impactor (NGI).

[0171] In one embodiment, the atomization time of the effective amount of 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 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 aminoglycoside formulation provided herein is approximately 13 minutes.

[0172] In one embodiment, the formulation described herein is administered once daily to a patient who requires it. [Examples]

[0173] The present invention will be further illustrated with reference to the following embodiments. However, it should be noted that these embodiments, like the embodiments described above, are illustrative and should not 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 formulations 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 Figure 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 by an amount in the range of 10 - 12 minutes (Figure 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 Figure 9 against the aerosol generation time. In other words, Figure 9 shows experimental data comparing the negative pressure ranges during aerosol generation for a reservoir (C) with a volume V RN of 24.5 mL and a reservoir (A) with 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 aerosol generator efficiency (proportional to liquid discharge rate or total discharge rate) on different negative pressures was measured using the nebulizer described above. Liposome amikacin formulations (thixotropes) with viscosities ranging from 5.5 to 14.5 mPa × s at shear forces of 1.1 to 7.4 Pa were 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 below approximately 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 the modified eFlow®. Here, the first aerosol generator (A) was used when the volume of the liquid reservoir increased V RN This is a modified eFlow® product with a volume of 19.5 mL, filled with 8 mL of liposomal amikacin formulation.

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

[0180] Figure 11 shows experimental data for these four aerosol generators filled with 8 mL of liposomal amikacin formulation. These results show that the aerosol generation time required to completely release the liposomal amikacin formulation from the reservoir is equal to the initial volume of liquid in the reservoir before use (V L ) when the volume of the liquid reservoir increases (V RN Figure 11 shows the relationship with the ratio of ) in the modified aerosol generator device (A), where an aerosol generation time of approximately 16 minutes was required, whereas the aerosol generation time was compared with the ratio V. RN / V LThis indicates a decrease as the amount of [unclear] increased. This data also shows that in the third aerosol generator device (C), the aerosol generation time could be reduced by approximately 4 minutes to less than 12 minutes.

[0181] Therefore, the data obtained in Example 1 shows that increasing the size of the air cushion allows the aerosol generator to operate in an efficient negative pressure range for a longer period of time, thereby significantly reducing the total aerosol generation time. Consequently, even a large amount of liquid, such as 8 mL, can be atomized (released in aerosol form) in less than 12 minutes. Example 2: Aerosol properties of amikacin formulations

[0182] Eleven liposomal amikacin formulations from different lots were examined using a modified eFlow® nebulizer (i.e., modified for use with the liposomal aminoglycoside formulations described herein), which had a modified 40-mesh membrane and a reservoir with an 8 mL liquid capacity and the air cushion described herein. Cascade impaction was performed using either an ACI (Andersen Cascade Impactor) or an NGI (Next Generation Impactor) to determine the aerosol properties, namely the aerodynamic median mass (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF). ACI measurement of aerodynamic median mass (MMAD)

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

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

[0185] The atomized material was collected in an 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 the weight difference (amount of atomization) divided by the time interval.

[0186] After collecting the sprayed material, ACI collection plates 0, 1, 2, 3, 4, 5, 6, and 7 were removed and placed in 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, and 10 mL for plates 0, 1, 5, 6, and 7) was added to each Petri dish. The samples from plates 0, 1, 2, 3, 4, 5, and 6 were further diluted appropriately with mobile phase C for HPLC analysis. The sample from plate 7 was analyzed by HPLC without further dilution. The ACI filters were also transferred to 20 mL vials, 10 mL of extraction solution was added, and all the formulation adhering to the vials was dissolved by vortexing the capped vials. The liquid samples from the vials were filtered (0.2 μm) into HPLC vials for HPLC analysis. The introduction port and connector were also rinsed with 10 mL of extraction solution to dissolve the formulation deposited therein, and the samples were collected and analyzed by HPLC at a 2-fold dilution. Based on the amount of amikacin deposited at each stage of the impactor, the aerodynamic median mass (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF) were calculated.

[0187] For nebulizers loaded with 8 mL and atomized for 6 minutes, the FPD (Flat Panel Quantity) was standardized relative to the volume of the atomized formulation in order to compare FPDs across all experiments. The FPD (standardized to the volume of the atomized formulation) was calculated according to the following equation.

number

[0188] Next-generation impactors (NGIs) were also used for MMAD measurements, and the atomization process was carried out inside a ClimateZone chamber (Westech Instruments Inc., Georgia) to maintain the temperature and RH% during atomization. The ClimateZone was preset to a temperature of 18°C ​​and a relative humidity of 50%. The NGI was assembled and loaded into the ClimateZone. A probe thermometer (VWR dual thermometer) was attached to the surface of the NGI to monitor its temperature. Atomization was initiated when the NGI temperature reached 18±0.5°C.

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

[0190] After aerosol collection, the NGI tray was removed from the NGI along with the tray holder. An appropriate amount of extract 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 then transferred to a volumetric flask. A 25 ml volumetric flask was used for NGI cups 1, 2, and 6, and a 50 ml volumetric flask was used for NGI cups 2, 3, and 4. Further extract solution was added to the cups and transferred again to the volumetric flask. This procedure was repeated several times to completely transfer the formulation deposited in the NGI cups to the volumetric flask. The volumetric flask was 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 diluted appropriately with mobile phase C for HPLC analysis. Samples from MOC were analyzed 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 of the adhering formulation. The liquid sample from the vial was filtered (0.2 microns) into an HPLC vial for HPLC analysis. The introduction port and connector were also rinsed with 10 mL of extraction solution to dissolve any deposited formulation, and the sample was collected and analyzed by HPLC at 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 across all experiments, FPD was standardized to the volume of the sprayed formulation. FPD (standardized to the volume of the sprayed formulation) was calculated according to the following equation.

number

[0193] The results of these experiments are presented in Figures 12 and 13, and in Table 5 below. [Table 5-1] [Table 5-2] Example 3: Atomization rate study

[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 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 volume after atomization was calculated as W3-W1. The atomization rate of the formulation was calculated according to the following formula:

number

[0195] Table 6 records the atomization rate (in g / min) and other relevant results for liposomal amikacin atomized using a nebulizer constructed according to this specification (24 aerosol heads were selected and used in these studies). [Table 6-1] [Table 6-2] [Table 6-3] Example 4: Percentage of Associated Amikacin After Atomization and Characterization of the Atomized Product

[0196] Free amikacin and liposome-encompassed amikacin were measured in the sprayed material of Example 3. As described in Example 3, the sprayed material was collected in a cooled impinger (Figure 14) at a flow rate of 8 L / min.

[0197] The sprayed material 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 to transfer all of the deposited preparation to the flask. To measure the free amikacin concentration of the sprayed material, 0.5 mL of the diluted sprayed material inside the volumetric flask was taken and placed in an Amicon® Ultra-0.5 mL 30K centrifuge filler. The sample was placed in a centrifuge (regenerated cellulose, 30K MWCO, Millipore) and centrifuged at 5000G and 15°C for 15 minutes. An appropriate amount of 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 using the following formula.

number

[0199] Table 7 summarizes the percentage of associated amikacin after atomization and the total dose recovery rate from the atomization experiments described in Table 6. 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 preparation was measured using the same HPLC and amikacin standard on the remaining samples. The obtained value was 64 mg / mL amikacin. The aggregated amikacin percentage after atomization ranged from 58.1% to 72.7%, with a mean of 65.5 ± 2.6%. When 8 mL of the liposomal amikacin preparation was atomized, the total amount of amikacin recovered ranged from 426 mg to 519 mg, with a mean of 476 ± 17 mg. The calculated amount of atomized amikacin (based on the weight of the atomized liposomal amikacin preparation in Table 7) ranged from 471 mg to 501 mg, with a mean of 490 ± 8 mg. The total amikacin recovery rate ranged from 91% to 104%, with a mean of 97 ± 3% (n=72). Liposome size

[0201] Liposome amikacin formulations (64 mg / mL amikacin) before or after atomization were appropriately diluted with 1.5% NaCl, and the liposome particle size was measured by light scattering using a Nicomp380 Submicron Particle Sizer (Nicomp, Santa Barbara, California).

[0202] The liposome size of aerosolized liposomal amikacin formulations was measured after atomization using 24 nebulizer aerosol heads in conjunction with an 8 mL reservoir handset. The liposome sizes ranged from 248.9 nm to 288.6 nm, with an average of 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 referenced herein are incorporated herein by reference in their entirety for all purposes.

[0204] The embodiments illustrated and described herein are intended only to teach those skilled in the art the best methods known to the inventors for carrying out and using the present invention. As will be understood in light of the above teachings, the above embodiments of the present invention can be modified and altered without departing from the present invention. Accordingly, it goes without saying that the present invention can be carried out in ways different from those specifically described, within the scope of the claims and their equivalents.

Claims

[Claim 1] The composition according to the specification.

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