System for treating pulmonary infection

Abstract

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

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority from U.S. Provisional Patent Application No. 61 / 649,830, filed May 21, 2012, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] 2. Background of the Invention Certain technologies suitable for administration by inhalation use liposomes, lipid complexes that provide a drug's therapeutic effect over a prolonged period in the lungs, and these technologies provide drugs with sustained activity and allow for targeting and enhanced uptake of the drug at disease sites.

[0003] Inhalation delivery of liposomes is complicated by the susceptibility of liposomes to shear-induced stresses upon nebulization that can result in changes in physical characteristics (e.g., encapsulation rate, size), but these do not necessarily pose an impediment to pharmaceutical development, so long as the changes in characteristics are reproducible and meet acceptance criteria.

[0004] Cystic fibrosis (CF) patients suffer from thick mucus and / or sputum secretions in the lungs, frequent secondary infections, and bacterial colonization of biofilms. All of these fluids and materials pose barriers to effective targeting of infections with aminoglycosides. Liposomal aminoglycoside formulations may help combat bacterial biofilms. Summary of the Invention [Means for solving the problem]

[0005] Summary of the Invention The present invention provides a method for treating various pulmonary infections, including mycobacterial infections, such as pulmonary infections caused by nontuberculous mycobacteria, also referred to herein as nontuberculous mycobacteria (NTM) infections, by providing a system for delivering an aerosolized liposomal formulation by inhalation. For example, the systems and methods provided herein can be used to treat pulmonary nontuberculous mycobacterial infections, such as pulmonary M. avium, M. avium, subsp.hominissuis (MAH), M.abscessus, M.chelonae, M.bolletii, M.kansasii, M.ulcerans, M.avium, M.avium complex (MAC) (M.avium and 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 aspect, the present invention provides a system for treating or preventing a pulmonary infection. In one embodiment, the system comprises a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, where the formulation is a dispersion (e.g., a liposomal solution or suspension) and the lipid component of the liposome is comprised of electrically neutral lipids, and a nebulizer that generates an aerosol of the pharmaceutical formulation at a rate of greater than about 0.53 g per minute. In one embodiment, the aerosol has a mass median aerodynamic diameter (MMAD) of less than about 4.2 μm as measured by an Anderson Cascade Impactor (ACI), between about 3.2 μm and about 4.2 μm as measured by an ACI, or less than about 4.9 μm as measured by a Next Generation Impactor (NGI), between about 4.4 μm and about 4.9 μm as measured by an NGI.

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

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

[0009] The pharmaceutical formulations provided herein are liposomal dispersions (i.e., liposomal dispersions or aqueous liposomal dispersions, which may be either liposomal solutions or liposomal suspensions). In one embodiment, the lipid components of the liposomes consist essentially of one or more electrically neutral lipids. In a further embodiment, the electrically neutral lipids comprise a phospholipid and a sterol. 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 formulation (aminoglycoside liposomal solution or suspension) is about 2:1, about 2:1 or less, about 1:1, about 1:1 or less, or about 0.7:1.

[0011] In one embodiment, the aerosolized aminoglycoside formulation, upon nebulization, has an aerosol droplet size 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. 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 nebulization. In a further embodiment, the aminoglycoside is selected from the aminoglycosides presented in Table A. In a further embodiment, the aminoglycoside is amikacin. In yet another embodiment, about 80% to about 100% of the amikacin is liposome complexed or about 80% to about 100% of the amikacin is encapsulated in multiple liposomes. In another embodiment, about 80% to about 100%, about 80% to about 99%, about 90% to about 100%, 90% to about 99%, or about 95% to about 99% of the aminoglycoside present in the formulation prior to nebulization is liposome complexed prior to nebulization.

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

[0014] In another aspect, the present invention provides a method for treating or preventing 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, such as a Psuedomonas aeruginosa infection. In another embodiment, the pulmonary infection is caused by one of the species of the genus Pseudomonas listed in Table B below. In one embodiment, a patient is treated for a mycobacterial pulmonary infection with one of the systems provided herein. In a further embodiment, the mycobacterial pulmonary infection is a nontuberculous mycobacterial pulmonary infection, a Mycobacterium abscessus pulmonary infection, or a Mycobacterium avium complex pulmonary infection. In one or more of the foregoing embodiments, the patient is a cystic fibrosis patient.

[0015] 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 caused by Mycobacterium abscessus, Mycobacterium avium complex, or P. 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. conpicuum, M. kansasii, M. peregrinum, M. immunogenum, M. xenopi, M. marinum, M. malmoense, M. marinum, M. mucogenicum, M. nonchromogenicum, M. scrofula. 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 aspect, a method is provided for treating or preventing a pulmonary infection in a patient. In one embodiment, the method comprises aerosolizing a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion of liposomes (e.g., a liposome solution or a liposome suspension) and is aerosolized at a rate greater than about 0.53 grams per minute. The method further comprises administering the aerosolized pharmaceutical formulation to the lungs of the patient, wherein the aerosolized pharmaceutical formulation comprises a mixture of free aminoglycoside and liposome-complexed aminoglycoside, and wherein the lipid component of the liposome is comprised of electrically neutral lipids. In a further embodiment, the aerosol has a mass median aerodynamic diameter (MMAD) of about 1.0 μm to about 4.2 μm as measured by ACI. In any one of the preceding embodiments, the aerosol has a MMAD of about 3.2 μm to about 4.2 μm as measured by ACI. In any one of the foregoing embodiments, the MMAD of the aerosol is from about 1.0 μm to about 4.9 μm as measured by NGI. In any one of the foregoing embodiments, the MMAD of the aerosol is from about 4.4 μm to about 4.9 μm as measured by NGI.

[0017] In one embodiment, the method comprises aerosolizing a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate of greater than about 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 aminoglycoside and liposome-complexed aminoglycoside (e.g., aminoglycoside encapsulated in liposomes), and the liposome component of the formulation is composed of electrically neutral lipids. In yet another embodiment, the fine particle fraction (FPF) of the aerosol is greater than or equal to about 64% as measured by ACI, or greater than or equal to about 51% as measured by NGI.

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

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

[0020] In one embodiment, the FPF of the lipid-complexed aminoglycoside aerosol is greater than or equal to about 64% as measured by an Andersen Cascade Impactor (ACI) or greater than or equal to about 51% as measured by a Next Generation Impactor (NGI). In a further embodiment, the aerosol comprises an aminoglycoside and a plurality of liposomes comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposomally complexed, e.g., encapsulated in a plurality of liposomes, and the liposomal aminoglycoside aerosol is generated at a rate greater than about 0.53 grams per minute. In any one of the foregoing embodiments, the aerosol is generated at a rate greater than about 0.54 grams per minute. In any one of the foregoing embodiments, the aerosol is generated at a rate greater than about 0.55 grams per minute. In any one of the foregoing embodiments, the aminoglycoside is selected from the aminoglycosides set forth in Table A.

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

[0022] In one embodiment, the concentration of the aminoglycoside in the liposome complexed aminoglycoside is about 50 mg / mL or greater. In a further embodiment, the concentration of the aminoglycoside in the liposome complexed aminoglycoside is about 60 mg / mL or greater. In a further embodiment, the concentration of the aminoglycoside in the liposome complexed aminoglycoside is about 70 mg / mL or greater, for example, about 70 mg / mL to about 75 mg / mL. In a further embodiment, the aminoglycoside is selected from the aminoglycosides presented in Table A. In yet another embodiment, the aminoglycoside is amikacin (e.g., amikacin sulfate). In an embodiment of the present invention, for example, the following items are provided: (Item 1) 1. A system for providing treatment or prophylaxis of a pulmonary infection in a patient, the system comprising: (a) a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, said formulation being an aqueous dispersion, and the lipid component of said liposomes being comprised of electrically neutral lipids; (b) a nebulizer that generates an aerosol of the pharmaceutical formulation at a rate greater than about 0.53 grams per minute; wherein the aerosol has a mass median aerodynamic diameter (MMAD) of less than about 4.2 μm as measured by an Andersen Cascade Impactor (ACI) or less than about 4.9 μm as measured by a Next Generation Impactor (NGI). (Item 2) 1. A system for providing treatment or prophylaxis for a pulmonary infection, the system comprising: (a) a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, said formulation being an aqueous dispersion, and the lipid component of said liposomes being comprised of electrically neutral lipids; (b) a nebulizer that generates an aerosol of the pharmaceutical formulation at a rate greater than about 0.53 grams per minute; wherein the fine particle fraction (FPF) of the aerosol is greater than or equal to about 64% as measured by ACI, or greater than or equal to about 51% as measured by NGI. (Item 3) 3. The system of claim 1 or 2, wherein the aminoglycoside is selected from amikacin, apramycin, arbekacin, astromycin, capreomycin, dibekacin, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamycin, or a combination thereof. (Item 4) The system according to any one of items 1 to 3, wherein the aminoglycoside is amikacin. (Item 5) The system according to any one of items 1 to 4, wherein the aminoglycoside is amikacin sulfate. (Item 6) 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 a sterol. (Item 8) The system according to any one of items 1 to 7, wherein the electrically neutral lipid comprises a phosphatidylcholine and a sterol. (Item 9) The system according to any one of items 1 to 8, wherein the electrically neutral lipid comprises dipalmitoylphosphatidylcholine (DPPC) and a sterol. (Item 10) The system according to any one of items 1 to 9, wherein the electrically neutral lipid comprises DPPC and cholesterol. (Item 11) 11. The system according to any one of items 1 to 10, wherein the aminoglycoside is amikacin, the electrically neutral lipid is composed of DPPC and cholesterol, and the liposome comprises a unilamellar vesicle, a multilamellar vesicle, or a mixture thereof. (Item 12) 12. The system according to any one of items 1 to 11, wherein the weight ratio of free aminoglycoside to said liposome-complexed aminoglycoside is from about 1:100 to about 100:1. (Item 13) 13. The system according to any one of items 1 to 12, wherein the weight ratio of free aminoglycoside to said liposome-complexed aminoglycoside is from about 1:10 to about 10:1. (Item 14) 14. The system according to any one of items 1 to 13, wherein the weight ratio of free aminoglycoside to said liposome-complexed aminoglycoside is from about 0.3:1 to about 2:1. (Item 15) 15. The system according to any one of items 1 to 14, wherein the volume of the pharmaceutical formulation is about 8 mL. (Item 16) 16. The system of any one of items 1 to 15, wherein the aerosol comprises about 55% to about 75% liposome-complexed amikacin. (Item 17) 17. The system according to any one of items 1 to 16, wherein the liposome-complexed aminoglycoside has an MMAD of about 3.2 μm to about 4.2 μm as measured by ACI, or about 4.4 μm to about 4.9 μm as measured by NGI. (Item 18) 18. The system according to any one of items 1 to 17, wherein the liposome-complexed aminoglycoside has an MMAD of about 3.6 μm to about 3.9 μm as measured by ACI, or about 4.5 μm to about 4.8 μm as measured by NGI. (Item 19) 19. The system of any one of items 1-18, wherein the FPF of the aerosolized formulation is greater than or equal to about 64% as measured by ACI, or greater than or equal to about 51% as measured by NGI. (Item 20) 20. The system of any one of items 1 to 19, wherein the FPF of the aerosolized formulation is about 64% to about 80% as measured by ACI, or about 51% to about 65% as measured by NGI. (Item 21) 21. The system of any one of items 1 to 20, wherein the nebulizer produces an aerosol of the pharmaceutical formulation at a rate of greater than about 0.54 g per minute. (Item 22) 22. The system of 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 pulmonary infection, and a liposome-complexed aminoglycoside in an amount effective to provide sustained bactericidal or sustained antibiotic activity against the pulmonary infection. (Item 23) 23. The system according to any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 500 mg to about 650 mg of an aminoglycoside. (Item 24) 23. The system according to any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 550 mg to about 625 mg of an aminoglycoside. (Item 25) 23. The system according to any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 550 mg to about 600 mg of an aminoglycoside. (Item 26) 23. The system according to any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 560 mg of an aminoglycoside. (Item 27) 23. The system according to any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 580 mg of an aminoglycoside. (Item 28) 23. The system according to any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 590 mg of an aminoglycoside. (Item 29) 23. The system according to any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 600 mg of an aminoglycoside. (Item 30) 1. A method for treating or preventing a pulmonary infection in a patient, the method comprising: aerosolizing a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein said pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate greater than about 0.53 grams per minute; and administering the aerosolized pharmaceutical formulation to the lungs of the patient. Including, the aerosolized pharmaceutical formulation comprises a mixture of free and liposome-complexed aminoglycosides; the lipid component of the liposome is composed of electrically neutral lipids; and The method, wherein the MMAD of the aerosol is less than about 4.2 μm as measured by ACI, or less than about 4.9 μm as measured by NGI. (Item 31) 31. The method according to item 30, wherein the MMAD of the aerosol is about 3.2 μm to about 4.2 μm when measured by ACI, or about 4.4 μm to about 4.9 μm when measured by NGI. (Item 32) 1. A method for providing treatment or prophylaxis for a pulmonary infection in a patient, the method comprising: aerosolizing a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein said pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate greater than about 0.53 grams per minute; and administering the aerosolized pharmaceutical formulation to the lungs of the patient. Including, the aerosolized pharmaceutical formulation comprises a mixture of free and liposome-complexed aminoglycosides; the lipid component of the liposome is composed of electrically neutral lipids; and The method, wherein the FPF of the aerosol is greater than or equal to about 64% as measured by ACI, or greater than or equal to about 51% as measured by NGI. (Item 33) 33. The method of any one of items 30-32, wherein the pharmaceutical formulation is aerosolized at a rate greater than about 0.55 grams per minute. (Item 34) 34. The method of any one of items 30 to 33, wherein the pharmaceutical formulation is aerosolized at a rate greater than about 0.56 grams per minute. (Item 35) 35. The method of any one of items 30 to 34, wherein the pharmaceutical formulation is aerosolized at a rate greater than about 0.58 grams per minute. (Item 36) 36. The method of any one of items 30 to 35, wherein the pharmaceutical formulation is aerosolized at a rate of about 0.60 to 0.80 grams per minute. (Item 37) 37. The method of any one of items 30-36, wherein the pharmaceutical formulation is aerosolized at a rate of about 0.60-0.70 grams per minute. (Item 38) 38. The method according to any one of items 30 to 37, wherein the pharmaceutical formulation comprises about 70 to about 75 mg / mL of amikacin, about 32 to about 35 mg / mL of DPPC, and about 16 to about 17 mg / mL of cholesterol. (Item 39) The method according to any one of items 30 to 38, wherein the pharmaceutical formulation has a volume of about 8 mL. 40. The method of any one of items 30 to 39, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astromycin, bekanamycin, boformycin, burramycin, capreomycin, dibekacin, dactimicin, etimicin, framycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomicin, ribostamycin, sisomicin, rhodostreptomycin, sorbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamycin, vertilmicin, or a combination thereof. (Item 41) 41. The method of any one of items 30 to 40, wherein the aerosolized pharmaceutical formulation is administered once daily in a single administration session. (Item 42) 42. The method according to any one of items 30 to 41, wherein the aminoglycoside is amikacin. (Item 43) 43. The method according to any one of items 30 to 42, wherein the aminoglycoside is amikacin sulfate. (Item 44) 44. The method of any one of items 30 to 43, wherein the free aminoglycoside is in an amount effective to provide immediate bactericidal or antibiotic activity against the nontuberculous mycobacterial infection and the liposome complexed aminoglycoside is in an amount effective to provide sustained bactericidal or antibiotic activity against the nontuberculous mycobacterial infection. (Item 45) 45. The method according to any one of items 30 to 44, wherein the pharmaceutical formulation comprises about 500 mg to about 650 mg of an aminoglycoside. (Item 46) 45. The method according to any one of items 30 to 44, wherein the pharmaceutical formulation comprises about 550 mg to about 625 mg of the aminoglycoside. (Item 47) 45. The method according to any one of items 30 to 44, wherein the pharmaceutical formulation comprises about 550 mg to about 600 mg of the aminoglycoside. (Item 48) 45. The method according to any one of items 30 to 44, wherein the pharmaceutical formulation comprises about 560 mg of the aminoglycoside. (Item 49) 45. The method according to any one of items 30 to 44, wherein the pharmaceutical formulation comprises about 580 mg of the aminoglycoside. (Item 50) 45. The method according to any one of items 30 to 44, wherein the pharmaceutical formulation comprises about 590 mg of the aminoglycoside. (Item 51) 45. The method according to any one of items 30 to 44, wherein the pharmaceutical formulation comprises about 600 mg of the aminoglycoside. (Item 52) nebulizing about 8 to about 9 grams of an aqueous liposomal dispersion of an aminoglycoside in less than about 16 minutes to obtain a liposomal aminoglycoside aerosol; and delivering said liposomal aminoglycoside aerosol to the patient's lungs by inhalation. 23. A method for delivering a liposomal aminoglycoside aerosol comprising: (Item 53) nebulizing about 8 to about 9 grams of an aqueous liposomal dispersion of aminoglycoside for about 10 to about 15 minutes to obtain a liposomal aminoglycoside aerosol; and delivering said liposomal aminoglycoside aerosol to the patient's lungs by inhalation. 23. A method for delivering a liposomal aminoglycoside aerosol comprising: (Item 54) 54. The method of claim 52 or 53, wherein the aqueous liposomal dispersion of aminoglycoside is nebulized in less than about 15 minutes, less than about 14 minutes, less than about 13 minutes, less than about 12 minutes, or less than about 11 minutes. (Item 55) 54. The method according to item 52 or 53, wherein the aqueous liposomal dispersion of aminoglycoside is nebulized in about 10 minutes to about 14 minutes, about 10 minutes to about 13 minutes, about 10 minutes to about 12 minutes, about 10 minutes to about 11 minutes, about 11 minutes to about 15 minutes, about 12 minutes to about 15 minutes, about 13 minutes to about 15 minutes, or about 14 minutes to about 15 minutes. (Item 56) 56. The method according to any one of items 52 to 55, wherein the liposomal aminoglycoside aerosol comprises an aminoglycoside and a liposome composed of DPPC and cholesterol, and about 55% to about 75% of the aminoglycoside is liposome-complexed. (Item 57) The liposomal aminoglycoside aerosol comprises: MMAD of about 3.2 μm to about 4.2 μm as measured by ACI, or about 4.4 μm to about 4.9 μm as measured by NGI; GSD of about 1.75 to about 1.80; An FPF greater than or equal to about 64% as measured by the ACI, or greater than or equal to about 51% as measured by the NGI; or Approximately 35 to 41 FPDs 57. The method according to any one of Items 52 to 56, comprising: (Item 58) 58. The method according to any one of items 52 to 57, wherein about 25% to about 35% of the liposomal aminoglycoside aerosol is deposited in the bronchial and alveolar regions of the patient's lungs. (Item 59) 59. The method of any one of items 52 to 58, wherein the aqueous liposomal dispersion of aminoglycoside comprises an aminoglycoside and liposomes composed of DPPC and cholesterol, and greater than about 95% of the aminoglycoside is encapsulated in the liposomes. (Item 60) 60. The method according to any one of items 52 to 59, wherein the aqueous liposomal dispersion of aminoglycoside contains about 70 to about 75 mg / mL of aminoglycoside, about 32 to about 35 mg / mL of DPPC, and about 16 mg / mL to about 17 mg / mL of cholesterol. (Item 61) 61. The method according to any one of items 52 to 60, wherein the volume of the aqueous liposome dispersion is about 8 mL. (Item 62) 62. The method of any one of items 52 to 61, wherein the aminoglycoside is amikacin. (Item 63) 63. The method of any one of items 52 to 62, wherein the aminoglycoside is amikacin sulfate. (Item 64) 64. The method of any one of items 52 to 63, wherein about 500 mg to about 650 mg of the aminoglycoside is delivered to the patient's lungs by inhalation. (Item 65) 64. The method of any one of items 52 to 63, wherein about 550 mg to about 625 mg of the aminoglycoside is delivered to the patient's lungs by inhalation. (Item 66) 64. The method of any one of items 52 to 63, wherein about 550 mg to about 600 mg of the aminoglycoside is delivered to the patient's lungs by inhalation. (Item 67) 64. The method of any one of items 52 to 63, wherein about 560 mg of the aminoglycoside is delivered to the patient's lungs by inhalation. (Item 68) 64. The method of any one of items 52 to 63, wherein about 580 mg of the aminoglycoside is delivered to the patient's lungs by inhalation. (Item 69) 64. The method of any one of items 52 to 63, wherein about 590 mg of the aminoglycoside is delivered to the patient's lungs by inhalation. (Item 70) 64. The method of any one of items 52 to 63, wherein about 600 mg of the aminoglycoside is delivered to the patient's lungs by inhalation. (Item 71) A liposomal aminoglycoside aerosol comprising an aminoglycoside and a liposome comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposomally complexed, and the liposomal aminoglycoside aerosol is produced at a rate greater than about 0.53 grams per minute. (Item 72) 72. The liposomal aminoglycoside aerosol according to item 71, which is produced at a rate greater than or equal to about 0.54 grams per minute, greater than or equal to about 0.55 grams per minute, or greater than or equal to about 0.60 grams per minute. (Item 73) 73. The liposomal aminoglycoside aerosol according to item 71 or 72, produced at a rate of about 0.60 to about 0.70 grams per minute. (Item 74) 74. The liposomal aminoglycoside aerosol of any one of items 71 to 73, wherein a single administration session provides about 500 mg or about 560 mg of amikacin. (Item 75) 75. The liposomal aminoglycoside aerosol of item 74, wherein the administration session is once a day. (Item 76) 76. The liposomal aminoglycoside aerosol according to any one of items 71 to 75, wherein the MMAD of the aerosol is less than about 4.2 μm as measured by ACI or less than about 4.9 μm as measured by NGI. (Item 77) 77. The liposomal aminoglycoside aerosol according to any one of items 71 to 76, wherein the MMAD of the aerosol is about 1.0 μm to about 4.2 μm when measured by ACI, about 2.0 μm to about 4.2 μm when measured by ACI, about 3.2 μm to about 4.2 μm when measured by ACI, about 1.0 μm to about 4.9 μm when measured by NGI, about 2.0 μm to about 4.9 μm when measured by NGI, and about 4.4 μm to about 4.9 μm when measured by NGI. (Item 78) 78. The liposomal aminoglycoside aerosol according to any one of items 71 to 77, wherein the fine particle fraction (FPF) of the aerosol is greater than or equal to about 64% as measured by ACI, or greater than or equal to about 51% as measured by NGI. (Item 79) 79. The liposomal aminoglycoside aerosol of any one of items 71 to 78, wherein the aerosol comprises greater than or equal to about 50% liposomal complexed amikacin. (Item 80) 80. The liposomal aminoglycoside aerosol of any one of items 71 to 79, wherein the aerosol comprises greater than or equal to about 60% liposomal complexed amikacin. (Item 81) 81. The liposomal aminoglycoside aerosol of any one of items 71 to 80, wherein the aerosol comprises about 55% to about 85% liposomal complexed amikacin. (Item 82) 82. The liposomal aminoglycoside aerosol according to any one of items 71 to 81, wherein the aerosol comprises about 55% to about 75% liposomal complexed amikacin. (Item 83) 83. The liposomal 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) 84. The liposomal aminoglycoside aerosol according to any one of items 71 to 83, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astromycin, bekanamycin, boformycin, burramycin, capreomycin, dibekacin, dactimicin, etimicin, framycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomicin, ribostamycin, sisomicin, rhodostreptomycin, sorbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamycin, vertilmicin, or a combination thereof. (Item 85) A liposomal aminoglycoside aerosol comprising an aminoglycoside and a plurality of liposomes comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposome-complexed, and the plurality of liposomes have a diameter of about 245 nm to about 290 nm as measured by light scattering. (Item 86) 86. The liposomal aminoglycoside aerosol according to item 85, produced at a rate of greater than about 0.53 grams per minute. (Item 87) 87. The liposomal aminoglycoside aerosol of item 85 or 86, wherein one administration session provides about 560 mg of amikacin. (Item 88) 88. The liposomal aminoglycoside aerosol according to any one of items 85 to 87, wherein the administration session is once a day. (Item 89) 89. The liposomal aminoglycoside aerosol according to any one of items 85 to 88, wherein the MMAD of the aerosol is from about 1.0 μm to about 4.2 μm when measured by ACI, from about 2.0 μm to about 4.2 μm when measured by ACI, from about 3.2 μm to about 4.2 μm when measured by ACI, from about 1.0 μm to about 4.9 μm when measured by NGI, from about 2.0 μm to about 4.9 μm when measured by NGI, and from about 4.4 μm to about 4.9 μm when measured by NGI. (Item 90) 90. The liposomal aminoglycoside aerosol according to any one of items 85 to 89, wherein the MMAD of the aerosol is from about 3.2 μm to about 4.2 μm when measured by ACI, or from about 4.4 μm to about 4.9 μm when measured by NGI. (Item 91) 91. The liposomal aminoglycoside aerosol according to any one of items 85 to 90, wherein the FPF of the aerosol is greater than or equal to about 64% as measured by ACI, or greater than or equal to about 51% as measured by NGI. (Item 92) 92. The liposomal aminoglycoside aerosol of any one of items 85-91, wherein the aerosol comprises greater than about 55% liposomal complexed amikacin, greater than about 60% liposomal complexed amikacin, greater than about 65% liposomal complexed amikacin, or greater than about 70% liposomal complexed amikacin. (Item 93) 93. The liposomal aminoglycoside aerosol of any one of items 85 to 92, wherein the aerosol comprises about 55% to about 75% liposomal complexed amikacin. (Item 94) 94. The liposomal aminoglycoside aerosol according to any one of items 85 to 93, wherein the liposome comprises a unilamellar vesicle, a multilamellar vesicle, or a mixture thereof. (Item 95) 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, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamycin, or a combination thereof. (Item 96) The liposomal aminoglycoside aerosol according to any one of items 71 to 95, which is produced at a rate of about 0.55 to about 0.70 grams per minute, or at a rate of about 0.60 to about 0.70 grams per minute, or at a rate of about 0.65 to about 0.70 grams per minute. (Item 97) 97. The liposomal aminoglycoside aerosol according to any one of items 85 to 96, wherein the average liposome size is about 265 nm as measured by light scattering. (Item 98) 52. The method of any one of items 30 to 51, wherein the patient has cystic fibrosis. (Item 99) 99. The method of any one of items 30-51 and 98, wherein the pulmonary infection is a nontuberculous mycobacterial infection. (Item 100) 99. The method of any one of items 30 to 51 and 98, wherein the pulmonary infection is a Pseudomonas infection. (Item 101) 99. The method of any one of items 30 to 51 and 98, wherein the pulmonary infection is a Burkholderia infection. (Item 102) 101. The method of item 100, wherein the Pseudomonas infection is a 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 of claim 99, wherein the nontuberculous mycobacterial infection is M. avium. 106. The method of claim 105, wherein the M. avium infection is a Mycobacterium avium subsp. hominissuis infection. (Item 107) 100. The method of claim 99, wherein the nontuberculous mycobacterial infection is a Mycobacterium abscessus infection. (Item 108) 100. The method of claim 99, wherein the nontuberculous mycobacterial infection is 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 99. The method of claim 99, wherein the fungus is selected from the group consisting of 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 a combination thereof. (Item 110) 30. The system according to any one of items 1 to 29, wherein the concentration of the aminoglycoside is about 50 mg / mL or more, or about 60 mg / mL or more, or about 70 mg / mL or more. (Item 111) 30. The system according to any one of items 1 to 29, wherein the concentration of the aminoglycoside is about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 76 mg / mL, about 77 mg / mL, about 78 mg / mL, or about 79 mg / mL. (Item 112) 30. The system according to any one of items 1 to 29, wherein the concentration of the aminoglycoside is about 60 mg / mL to about 80 mg / mL. (Item 113) The system according to any one of items 110 to 112, wherein the aminoglycoside is amikacin. (Item 114) Item 114. The system of item 113, wherein the aminoglycoside is amikacin sulfate. (Item 115) The method according to any one of items 30 to 70 and 98 to 109, wherein the concentration of the aminoglycoside is about 50 mg / mL or more, or about 60 mg / mL or more, or about 70 mg / mL or more. (Item 116) The method according to any one of items 30 to 70 and 98 to 109, wherein the concentration of the aminoglycoside is about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 76 mg / mL, about 77 mg / mL, about 78 mg / mL or about 79 mg / mL. (Item 117) The method according to any one of items 30 to 70 and 98 to 109, wherein the concentration of the aminoglycoside is about 60 mg / mL to about 80 mg / mL. (Item 118) The method according to any one of items 30 to 70 and 98 to 109, wherein the concentration of the aminoglycoside is about 70 mg / mL to about 80 mg / mL. (Item 119) 119. The method of any one of items 116 to 118, wherein the aminoglycoside is amikacin. (Item 120) 120. The method of claim 119, wherein the aminoglycoside is amikacin sulfate. (Item 121) 98. The liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is about 50 mg / mL or more, or about 60 mg / mL or more, or about 70 mg / mL or more. (Item 122) 98. The liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 76 mg / mL, about 77 mg / mL, about 78 mg / mL or about 79 mg / mL. (Item 123) 98. The liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is about 60 mg / mL to about 80 mg / mL. (Item 124) 98. The liposomal aminoglycoside aerosol according to any one of items 71 to 97, wherein the concentration of the aminoglycoside is about 70 mg / mL to about 80 mg / mL. (Item 125) 125. The liposomal aminoglycoside aerosol according to any one of items 122 to 124, wherein the aminoglycoside is amikacin. (Item 126) 126. The liposomal aminoglycoside aerosol according to item 125, wherein the aminoglycoside is amikacin sulfate. (Item 127) 30. The system of any one of items 1 to 29, wherein the pulmonary infection is a nontuberculous mycobacterial pulmonary infection. (Item 128) 30. The system according to any one of items 1 to 29, wherein the pulmonary infection is a Pseudomonas pulmonary infection. (Item 129) 30. The system of any one of items 1 to 29, wherein the pulmonary infection is a Burkholderia pulmonary infection. (Item 130) Item 129. The system of item 128, wherein the Pseudomonas infection is a 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 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 infection, or a combination thereof. (Item 133) Item 133. The system of item 132, wherein the nontuberculous mycobacterial pulmonary infection is an M. abscessus pulmonary infection. (Item 134) Item 133. The system of item 132, wherein the nontuberculous mycobacterial pulmonary infection is an M. avium pulmonary infection. (Item 135) Item 135. The system of item 134, wherein the nontuberculous mycobacterial pulmonary infection is an M. avium subsp. hominissuis pulmonary infection. (Item 137) 71. The method of any one of items 30-39, 41 and 44-70, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astromycin, bekanamycin, boformycin, burramycin, capreomycin, dibekacin, dactimicin, etimicin, framycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomicin, ribostamycin, sisomicin, rhodostreptomycin, sorbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamycin, vertilmicin, or a combination thereof. (Item 138) 115. The system of any one of items 1-3, 6-29 and 110-114, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astromycin, bekanamycin, boformycin, burramycin, capreomycin, dibekacin, dactimicin, etimicin, framycetin, gentamicin, H107, hygromycin, hygromycin B, inosamycin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomicin, ribostamycin, sisomicin, rhodostreptomycin, sorbistin, spectinomycin, sporaricin, streptomycin, tobramycin, verdamycin, vertilmicin, or a combination thereof. (Item 139) 139. The system according to any one of items 1 to 29, 110 to 114 and 138, wherein the aqueous dispersion is an aqueous suspension of a liposome-complexed aminoglycoside. (Item 140) 139. The system according to any one of items 1 to 29, 110 to 114 and 138, wherein the aqueous dispersion is an aqueous solution of a liposome-complexed aminoglycoside. (Item 141) 138. The method according to any one of items 30 to 70, 115 to 120 and 137, wherein the aqueous dispersion is an aqueous suspension of a liposome-complexed aminoglycoside. (Item 142) 138. The method according to any one of items 30 to 70, 115 to 120 and 137, wherein the aqueous dispersion is an aqueous solution of a liposome-complexed aminoglycoside. (Item 143) 99. The method of any one of items 30 to 51 and 98, wherein the pulmonary infection is associated with bronchiectasis. (Item 144) 144. The method of claim 143, wherein the aqueous dispersion is an aqueous suspension of a liposome-complexed aminoglycoside. (Item 145) 144. The method of claim 143, wherein the aqueous dispersion is an aqueous solution of a liposome-complexed aminoglycoside. (Item 146) 30. The system of any one of items 1 to 29, wherein the pulmonary infection is associated with bronchiectasis. [Brief description of the drawings]

[0023] [Figure 1] FIG. 1 is a diagram of a nebulizer (aerosol generator) in which the present invention can be implemented.

[0024] [Diagram 2] FIG. 2 is an enlarged view of the nebulizer diagram shown in FIG.

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

[0026] [Figure 4]FIG. 4 is an image of a PARI eFlow® nebulizer modified for use with the aminoglycoside formulations described herein, and a blown-up view of the nebulizer membrane.

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

[0028] [Figure 6] FIG. 6 is a computed tomography (CT) cross-sectional image of a stainless steel membrane having a relatively short nozzle portion.

[0029] [Figure 7] FIG. 7 is a schematic cross-sectional view of a sputum / biofilm, such as may be found in a patient with cystic fibrosis.

[0030] [Figure 8] FIG. 8 is a graph of the duration of aerosol generation (nebulization time) upon complete release of the liquid in the reservoir as a function of the initial gas cushion (VA) in the reservoir.

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

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

[0033] [Figure 11] FIG. 11 is a graph of the duration for aerosol generation (nebulization time) upon complete release of liquid as a function of the ratio (VRN / VL) between the increased volume of the reservoir VRN and the initial volume of liquid in the reservoir (VL).

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

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

[0036] [Figure 14] FIG. 14 is a schematic diagram of the system used for aerosol collection for post-nebulization studies. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0037] Detailed Description of the Invention The invention described herein is directed, in part, to a system for administering an aminoglycoside pharmaceutical formulation to the lungs of a subject, for example, to treat a pulmonary disorder.

[0038] The term "treating" includes (1) preventing or delaying the appearance of clinical symptoms of the situation, disorder or condition in a subject who may be afflicted with or predisposed to the situation, disorder or condition, but who has not yet experienced or exhibited clinical or subclinical symptoms of the situation, disorder or condition; (2) inhibiting the situation, disorder or condition (i.e., arresting, reducing or delaying the onset of at least one clinical or subclinical symptom of the disease, or, in the case of maintenance treatment, of a flare-up); and / or (3) alleviating the condition (i.e., causing regression of the situation, disorder or condition, or at least one clinical or subclinical symptom thereof). The benefit to a subject being treated is statistically significant or at least perceptible to the subject or to the physician.

[0039] In one embodiment, the systems and formulations provided herein can treat lung infections caused by the following bacteria: Pseudomonas (e.g., P. aeruginosa, P. paucimobilis, P. putida, P. fluorescens, and P. acidovorans), Burkholderia (e.g., B. pseudomallei, B. cepacia, B. cepacia complex, B. dolosa, B. fungorum, B. gladioli, B. multivulgare, B. erythropoeias ... 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 Staphylococcus aureus (MRSA), Streptococcus (e.g. Streptococcus pneumoniae), Escherichia coli, Klebsiella, Enterobacter, Serratia, Haemophilus, Yersinia pestis, Mycobacterium (e.g. nontuberculous mycobacteria).

[0040] In one embodiment, a patient is treated for a nontuberculous mycobacterial lung infection with 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 presented in Table B below. [Table B-1] [Table B-2]

[0042] The nontuberculous mycobacterial lung infection, in one embodiment, is selected from the group consisting of 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. conpicuum, M. kansasii, M. peregrinum, M. immunogenum, M. xenopi, M. marinum, M. malmoense, M. marinum, M. mucogenicum, M. nonchromogenicu 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 a refractory nontuberculous mycobacterial lung infection.

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

[0044] As used herein, "prophylaxis" can mean preventing an infection or disease altogether, or preventing the onset of a symptom of that infection or disease; delaying the onset of an infection or disease or a symptom thereof; or reducing the severity of a later occurring infection or disease or a symptom thereof.

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

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

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

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

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

[0050] The therapeutic response can be any response that a user (e.g., a clinician) will recognize as an effective response to treatment. The therapeutic response will generally be the reduction, inhibition, delay or prevention of the growth or proliferation of one or more of the above-mentioned bacteria, or the killing of one or more of the above-mentioned bacteria. The therapeutic response can also be reflected in the improvement of lung function, such as forced expiratory volume in 1 second (FEV1). Furthermore, it is within the skill of the person skilled in the art to determine the appropriate treatment duration, appropriate dosage, and any potential concomitant treatment based on the evaluation of the therapeutic response.

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

[0052] As used herein, an "aerosol" is a gas phase suspension of liquid particles. Aerosols provided herein include particles of liposomal 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 respiratory tract. Pneumotic 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, provided that the particular nebulizer delivers an aerosol of the required properties at the required output rate.

[0054] The process of converting bulk liquid into small droplets with air is called nebulization. Pneumatic nebulizers require a pressurized gas supply to operate as the driving force for liquid atomization. Ultrasonic nebulizers use electricity introduced by a piezoelectric element in a liquid reservoir to convert liquid into respirable droplets. Various types of nebulizers are described in Respiratory Care, Vol. 45, No. 6, pp. 609-622 (2000), the disclosure of which is incorporated herein by reference in its entirety. The terms "nebulizer" and "aerosol generator" are used interchangeably throughout this specification. The terms "inhalation device," "inhalation system," and "atomizer" are also used interchangeably in the literature with the terms "nebulizer" and "aerosol generator."

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

[0056] "Mass Median Diameter" or "MMD" is the average particle size on a mass basis as determined by laser diffraction or impactor measurements.

[0057] "Mass Median Aerodynamic Diameter" or "MMAD" is standardized for the aerodynamic separation of aqueous aerosol droplets and is determined by impactor measurements, such as an Andersen Cascade Impactor (ACI) or Next Generation Impactor (NGI). The gas flow rate, in one embodiment, is 28 liters per minute for an Andersen Cascade Impactor (ACI) and 15 liters per minute for a Next Generation Impactor (NGI). "Geometric Standard Deviation" or "GSD" is a measure of the spread of the aerodynamic particle size distribution.

[0058] In one embodiment, the present invention provides a system for treating or preventing a pulmonary infection. Treatment is achieved via delivery of an aminoglycoside formulation by inhalation via nebulization. In one embodiment, the pharmaceutical formulation comprises an aminoglycoside agent, e.g., an aminoglycoside.

[0059] The pharmaceutical formulations provided herein are liposomal dispersions. Specifically, the pharmaceutical formulations are dispersions that include "liposome-complexed aminoglycosides" or "liposomally encapsulated aminoglycosides." "Liposome-complexed aminoglycosides" encompasses embodiments in which the aminoglycoside (or combination of aminoglycosides) is encapsulated in liposomes, and encompasses any form of aminoglycoside composition in which at least about 1% by weight of the aminoglycoside is associated with liposomes, either as part of a complex with liposomes or as liposomes in which the aminoglycoside may be in the aqueous or hydrophobic bilayer phase, or in the interfacial headgroup region of the liposomal bilayer.

[0060] In one embodiment, the lipid component of the liposome comprises electrically neutral lipids, positively charged lipids, negatively charged lipids, or a combination thereof. In another embodiment, the lipid component comprises electrically neutral lipids. In a further 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 set forth above, liposome-complexed aminoglycoside embodiments encompass those in which the aminoglycoside is encapsulated in a liposome. In addition, liposome-complexed aminoglycoside refers to any composition, solution, or suspension in which at least about 1% by weight of the aminoglycoside is associated with lipids, either as part of a complex with a liposome, or as a liposome in which the aminoglycoside may be in the aqueous phase, the hydrophobic bilayer phase, or at the interfacial headgroup region of the liposome bilayer. In one embodiment, at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% of the aminoglycoside in the formulation is so associated prior to nebulization. Association, in one embodiment, is measured by separation through a filter, such that the lipid and lipid-associated drug are retained (i.e., retained in the retentate) and the free drug enters the filtrate.

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

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

[0064] In one embodiment, the pharmaceutical formulation comprises dipalmitoylphosphatidylcholine (DPPC), a major component of natural pulmonary surfactant. In one embodiment, the lipid component of the pharmaceutical formulation comprises, consists essentially of, or consists of DPPC and cholesterol. In a further embodiment, the DPPC and cholesterol have a molar ratio ranging from about 19:1 to about 1:1, or from about 9:1 to about 1:1, or from about 4:1 to about 1:1, or from about 2:1 to about 1:1, or from about 1.86:1 to about 1:1. In yet another embodiment, the DPPC and cholesterol have a molar ratio of about 2:1 or about 1:1. In one embodiment, the DPPC and cholesterol are provided in an aminoglycoside formulation, e.g., an aminoglycoside formulation.

[0065] Other examples of lipids for use with the present invention include, but are not limited to, dimyristoylphosphatidychloline (DMPC), dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylcholine (DSPC), distearoylphosphatidylglycerol (DSPG), dioleoylphosphatidyl-ethanolamine (DOPE), mixed phospholipids such as palmitoylstearoylphosphatidyl-choline (PSPC), and monoacylated phospholipids such as monooleoyl-phosphatidylethanolamine (MOPE).

[0066] In one embodiment, said at least one lipid component comprises a sterol. In a further embodiment, said at least one lipid component comprises a sterol and a phospholipid, or consists essentially of a sterol and a phospholipid, or consists of a sterol and a phospholipid. The sterols used with the present invention include, but are not limited to, cholesterol, esters of cholesterol, such as cholesterol hemisuccinate, salts of cholesterol, such as cholesterol hydrogen sulfate and cholesterol sulfate, ergosterol, esters of ergosterol, such as ergosterol hemisuccinate, salts of ergosterol, such as ergosterol hydrogen sulfate and ergosterol sulfate, lanosterol, esters of lanosterol, such as lanosterol hemisuccinate, salts of lanosterol, such as lanosterol hydrogen sulfate, lanosterol sulfate, and tocopherol. Tocopherols can include tocopherol, esters of tocopherol such as tocopherol hemisuccinate, salts of tocopherol such as tocopherol hydrogen sulfate and tocopherol sulfate. The term "sterol compound" includes 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 can include ammonium salts of fatty acids, phospholids, and glycerides. The fatty acids include fatty acids with carbon chain lengths of 12 to 26 carbon atoms that are saturated or unsaturated. Some specific examples include: myristylamine, palmitylamine, laurylamine, and stearylamine, dilauroylethylphosphocholine (DLEP), dimyristoylethylphosphocholine (DMEP), dipalmitoylethylphosphocholine (DPEP), and distearoylethylphosphocholine (DSEP), N-(2,3-di-(9-(Z)-octadecenyloxy)-prop-1-yl-N,N,N-trimethylammonium chloride (DOTMA), and 1,2-bis(oleoyloxy)-3-(trimethylammonio)propane (DOTAP).

[0068] In one embodiment, at least one anionic lipid (negatively charged lipid) is provided in the system described herein.The negatively charged lipid that can be used includes 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] Without wishing to be bound by theory, phosphatidylcholine such as DPPC aids in the uptake of aminoglycosides by cells in the lung (e.g., alveolar macrophages) and helps maintain aminoglycosides in the lung. Negatively charged lipids such as PG, PA, PS and PI are believed 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. Without wishing to be bound by theory, sterol compounds are believed to affect the release characteristics of the formulation.

[0070] Liposomes are completely closed lipid bilayer membranes that contain a certain amount of encapsulated water. Liposomes can be unilamellar vesicles (having a single membrane bilayer) or multilamellar vesicles (onion-like structures characterized by multiple membrane bilayers, each separated from the next by an aqueous layer), or a combination thereof. The bilayer is composed of two lipid monolayers with hydrophobic "tail" regions and hydrophilic "head" regions. The structure of the membrane bilayer is such that the hydrophobic (non-polar) "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 made by a variety of methods (see, for example, Cullis et al. (1987)). In one embodiment, one or more of the methods described in US Patent Application Publication No. 2008 / 0089927 are used herein to make aminoglycoside-encapsulated lipid formulations (liposomal dispersions). The disclosure of US Patent Application Publication No. 2008 / 0089927 is incorporated herein by reference in its entirety for all purposes. For example, in one embodiment, at least one lipid and an aminoglycoside are mixed with a coacervate (i.e., a separate liquid phase) to form a liposomal formulation. The coacervate can be formed prior to mixing with the lipid, during mixing with the lipid, or after mixing with the lipid. The coacervate can also be a coacervate of an active agent.

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

[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 conventional multilamellar vesicles (MLVs). Lenk et al. (U.S. Pat. Nos. 4,522,803, 5,030,453, and 5,169,637), Fountain et al. (U.S. Pat. No. 4,588,578), and Cullis et al. (U.S. Pat. No. 4,975,282) disclose methods for producing multilamellar liposomes with substantially equal interlamellar solute distribution in each of the aqueous compartments. Paphadjopoulos et al. (U.S. Pat. No. 4,235,871) discloses the preparation of oligolamellar liposomes by reverse phase evaporation. Each of these methods is suitable for use with the present invention.

[0074] Unilamellar vesicles can be produced from MLVs by several techniques, such as the extrusion technique of U.S. Patent Nos. 5,008,050 and 5,059,421. Sonication and homogenization can also be used to produce small unilamellar liposomes from larger liposomes (see, e.g., Paphadjopoulos et al. (1968); Deamer and Uster (1983); and Chapman et al. (1968)).

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

[0076] Liposomes for use in the pharmaceutical formulations provided herein can be prepared using techniques such as reverse phase evaporation, infusion method, and detergent dilution to prepare large unilamellar vesicles (LUVs).Reviews of these and other methods for preparing liposomes can be found in Chapter 1 of the book "Liposomes," edited by Marc Ostro, Marcel Dekker, Inc., New York, 1983, which is incorporated herein by reference.See also Szoka, Jr. et al. (Ann.Rev.Biophys.Bioeng.9,1980,p.467), which is incorporated herein by reference for all purposes.

[0077] Other techniques for making liposomes include forming reverse phase evaporation vesicles (REV), U.S. Patent No. 4,235,871. Another type of liposome that can be used is characterized by having a substantially equal lamellar solute distribution. This type of liposome is called stable plurilamellar vesicles (SPLV) 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 (FATMLV) described above.

[0078] Liposomes have been formed using various sterols and their water-soluble derivatives, such as cholesterol hemisuccinate. See, e.g., U.S. Patent No. 4,721,612. Mayhew et al., PCT Publication No. WO 85 / 00968, describe a method for reducing the toxicity of drugs by encapsulating the drugs in liposomes containing α-tocopherol and certain derivatives thereof. Liposomes have also been formed using various tocopherols and their water-soluble derivatives. See, PCT Publication No. 87 / 02219.

[0079] In one embodiment, the pharmaceutical formulation comprises liposomes having an average diameter, measured by light scattering, in the range of about 0.01 microns to about 3.0 microns, e.g., about 0.2 to about 1.0 microns, prior to nebulization. In one embodiment, the average diameter of the liposomes in the formulation is about 200 nm to about 300 nm, about 210 nm to about 290 nm, about 220 nm to about 280 nm, about 230 nm to about 280 nm, about 240 nm to about 280 nm, about 250 nm to about 280 nm, or about 260 nm to about 280 nm. The sustained activity profile of the liposomal product can be controlled by the properties of the lipid membrane and by including other excipients in the composition.

[0080] To minimize the volume administered and reduce patient administration time, in one embodiment, it is important that liposomal encapsulation of the aminoglycoside (e.g., the aminoglycoside amikacin) be extremely efficient and that the L / D ratio be as low as possible and / or practical, while keeping the liposomes small enough to penetrate the patient's 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 liposomes have an average diameter of about 260 to about 280 nm, as measured by light scattering.

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

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

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

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

[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 a formulation as shown in Table 2 below. [Table 2]

[0086] It should be noted that increasing the aminoglycoside concentration alone will not result in a reduction in administration time. For example, in one embodiment, the lipid to drug ratio is fixed, and as 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 nebulization time.

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

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

[0089] As provided herein, the present invention provides methods and systems for treating pulmonary infections by inhalation of a liposomal aminoglycoside formulation via nebulization. In one embodiment, the formulation is administered via a nebulizer that provides an aerosol mist of the formulation for delivery to the subject's lungs.

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

[0091] Upon nebulization, the liposomes in the pharmaceutical formulation leak the drug. In one embodiment, the amount of liposome-complexed aminoglycoside after nebulization 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 "associated aminoglycoside percentage after nebulization." As provided herein, in one embodiment, the liposomes include an aminoglycoside, such as amikacin. In one embodiment, the associated aminoglycoside percentage after nebulization is about 60% to about 70%. In a further embodiment, the aminoglycoside is amikacin. In another embodiment, the associated aminoglycoside percentage after nebulization is about 67%, or about 65% to about 70%. In a further embodiment, the aminoglycoside is amikacin.

[0092] In one embodiment, the percentage associated aminoglycoside after nebulization is measured by recovering the aerosol from the air by condensation in a cold trap, and the liquid is then assayed for free 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 as measured by ACI at a gas flow rate of about 28 L / min or Next Generation Impactor NGI at a gas flow rate of about 15 L / min.

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

[0095] In another embodiment, a nebulizer as described herein produces an aerosol of the aminoglycoside pharmaceutical formulation at a rate of greater than about 0.53 g / min, greater than about 0.55 g / min, or greater than about 0.60 g / min, or between about 0.60 g / min and about 0.70 g / min. In a further embodiment, the FPF of the aerosol is greater than or equal to about 64% as measured by ACI, greater than or equal to about 70% as measured by ACI, greater than or equal to about 51% as measured by NGI, or greater than or equal to about 60% as 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 working principle of pneumonic nebulizers is well known to those skilled in the art and is described, for example, in Respiratory Care, Vol. 45, No. 6, pp. 609-622 (2000). Briefly, pneumonic nebulizers use a pressurized gas supply as the driving force for liquid atomization. Compressed gas is delivered, which creates a region of negative pressure. The solution to be aerosolized is then delivered into the gas stream and sheared into a liquid film. This film is unstable and breaks into droplets due to surface tension. Baffles can then be placed in the aerosol stream to form smaller particles, i.e., particles with the MMAD and FPF properties described above. In one embodiment of a pneumonic nebulizer, the gas and solution are mixed before leaving the exit port (nozzle) and interacting with the baffle. In another embodiment, mixing does not occur until the liquid and gas leave the exit 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 in a pneumonic nebulizer. However, consideration should be given to the formulation to be nebulized and whether the properties of the formulation (e.g., % associated aminoglycoside) are changed by adjusting the nebulizer. For example, in one embodiment, the gas velocity and / or pharmaceutical formulation velocity are adjusted to achieve the discharge rate and droplet size of the present invention. Additionally or alternatively, 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, an increase in gas velocity, 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, an increase in the ratio of liquid flow rate to gas flow rate increases the particle size.

[0099] In one embodiment, the discharge rate of a pneumonic nebulizer is increased by increasing the fill volume in the reservoir. Without wishing to be bound by theory, the increased discharge rate may be due to a reduction in the dead volume in the nebulizer. In one embodiment, the nebulization time is reduced by increasing the flow rate driving the nebulizer. See, e.g., 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 nebulization process, which is then provided to the subject via inhalation. In another embodiment, the nebulizer provided herein includes a valved open vent design. In this embodiment, as the patient inhales through the nebulizer, the output volume of the nebulizer increases. During the exhalation phase, a one-way valve redirects the patient flow away from the nebulizer chamber.

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

[0102] In one embodiment, a vibrating mesh nebulizer is used to deliver the aminoglycoside formulation of the present invention to a patient in need thereof. In one embodiment, the nebulizer membrane vibrates at an ultrasonic frequency of about 100 kHz to about 250 kHz, about 110 kHz to about 200 kHz, about 110 kHz to about 200 kHz, or about 110 kHz to about 150 kHz. In one embodiment, upon application of electrical current, the nebulizer membrane vibrates at a frequency of about 117 kHz.

[0103] In one embodiment, the nebulizer provided herein does not use an air compressor and therefore does not generate an 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 at the back of the mixing chamber and carries the aerosol to the patient through the mouthpiece. When exhaling, 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 on the next breath. This cycle continues until the nebulizer's dosage reservoir is empty.

[0104] The present invention, in one aspect, is practiced using, but is not limited to, one of the aerosol generators (nebulizers) depicted in Figures 1, 2, 3 and 4. The system of the present invention also includes, in one embodiment, a nebulizer as described in European Patent Application Nos. 11169080.6 and / or 10192385.2, both of which are incorporated herein by reference in their entireties.

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

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

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

[0108] In one embodiment, the length of the nozzle portion of the through-hole formed in the membrane (e.g., the vibrable membrane) affects the total output 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 output velocity, the shorter the nozzle portion, the higher the 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, hi a further embodiment, the length of the portion of the through-hole upstream of the nozzle portion does not have a significant effect on the TOR.

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

[0111] In one embodiment, as set forth above, the systems provided herein include a nebulizer that produces an aerosol of the aminoglycoside pharmaceutical formulation at a rate greater than about 0.53 g per minute, or greater than about 0.55 g per minute. In a further embodiment, the nebulizer includes a vibratable membrane having a first surface in contact with the fluid and an opposing second surface from which the droplets emerge.

[0112] The membrane, e.g., a stainless steel membrane, can be vibrated using a piezoelectric actuator or any other suitable means. The membrane has a plurality of through-holes extending through it in a direction of extension from the first surface to the second surface. The through-holes can be formed as described above by a laser source, electroforming, or any other suitable process. As the membrane is vibrated, the aminoglycoside pharmaceutical formulation passes through the through-holes from the first surface to the second surface, thereby generating an aerosol on the second surface. In one embodiment, each of the through-holes includes an inlet opening and an outlet opening. In a further embodiment, each of the through-holes includes a nozzle portion extending from the outlet opening 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 direction of extension that includes the minimum diameter of the through-hole and is bounded by the larger diameter of the through-hole. In one embodiment, the larger diameter of the through-hole is defined as the diameter that is 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 smallest diameter of the through-hole is the diameter of the exit opening, hi another embodiment, the smallest diameter of the through-hole is about 0.5x, about 0.6x, about 0.7x, about 0.8x, or about 0.9x the diameter of the exit opening.

[0114] In one embodiment, the nebulizers provided herein include through-holes having a ratio of the total length in an elongated direction of at least one through-hole to the length in an elongated direction of each nozzle portion of the through-hole of 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 nebulizers provided herein include through-holes having a ratio of the total length in an elongated direction of a majority of the through-holes to the length in an elongated direction of each nozzle portion of the through-hole of 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 above-described extension ratios, in one embodiment, provide an increased total delivery rate and also provide sufficient GSD compared to known nebulizers. These ratio configurations, in one embodiment, shorten the application 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 a low concentration due to its nature, and therefore a larger volume of the aminoglycoside pharmaceutical formulation must be administered within an acceptable time, such as one dosing session.

[0116] According to one embodiment, the nozzle portion ends flush with the second surface. The length of the nozzle portion is therefore defined in one embodiment as the portion beginning at the second surface toward the first surface and bounded by a diameter that is closest to about 3 times, about 2 times, about 2.5 times, or about 1.5 times the minimum diameter. The minimum diameter in this embodiment is the diameter of the outlet opening.

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

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

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

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

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

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

[0123] Alternatively or additionally, the through hole is formed by an electroforming process. In one embodiment, the through hole has a first funnel-shaped portion on the first surface and a second funnel-shaped portion on the second surface, with a nozzle portion between the first and second funnel-shaped portions and defined between the outlet opening and the larger diameter. In this case, the overall length of the through hole may also be defined only by the distance from the first surface to the outlet opening (smallest diameter).

[0124] In addition, the total discharge rate (TOR) can be further increased by increasing the number of through-holes provided in the membrane. In one embodiment, this is achieved by increasing the active perforation surface of the membrane and keeping the distance between the through-holes at the same level. In another embodiment, the number of through-holes is increased by decreasing the distance between the through-holes and maintaining the active area of ​​the membrane. In addition, a combination of the above strategies may also be used.

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

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

[0127] 3 shows an aerosol generator (nebulizer) as disclosed in WO2001 / 032246, the entirety of which is incorporated herein by reference. The aerosol generator comprises 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 using a mouthpiece 4 via an opening 41.

[0128] The aerosol generator comprises a vibratable membrane 22 which is vibrated using a piezoelectric actuator 23. The vibratable membrane 22 has a first surface 24 facing the fluid container 21 and a second, opposite surface 25 facing the mixing chamber 3. In use, the first surface 24 of the vibratable membrane 22 is in contact with the fluid contained in the fluid container 21. The membrane 22 is provided with a number of through holes 26 which pass through the membrane from the first surface 24 to the second surface 25. In use, fluid passes from the fluid container 21 through the through holes 26 from the first surface 24 to the second surface 25, causing the membrane 22 to vibrate in order to generate an aerosol at the second surface 25 and release it into the mixing chamber 3. The aerosol can then be drawn from the mixing chamber 3 by inhalation by the patient via the mouthpiece 4 and its inhalation opening 41.

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

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

[0131] Both vibrable membranes of Figures 5 and 6 were fabricated with 6,000 through holes 26. The table below (Table 3) shows the mass median diameter (MMD) determined by laser diffraction, the time required to completely expel a volume of liquid (nebulization time), and the TOR of particles expelled on the second side of the membrane. Tests were performed with a liposomal formulation of amikacin. [Table 3]

[0132] Table 3 shows that Membrane 2, with its short nozzle section, provides increased TOR and a nebulization time that is approximately 36% less, or 5.3 minutes, shorter, compared to Membrane 1. Table 3 also shows that the MMD did not vary significantly for each membrane tested, which is in contrast to the differences in TOR observed for each membrane. Thus, in one embodiment, the nebulizer described herein provides a significant reduction in nebulization time compared to prior art nebulizers, without affecting droplet size as measured by MMD.

[0133] In addition to the membranes shown in Figures 5 and 6, membranes were also fabricated with a further reduced nozzle section and 3000 through holes 26 (Membranes 3 and 4 in Table 3). Specifically, Membrane 3 was laser drilled to have a shorter nozzle section, and Membrane 4 was fabricated to have a shorter nozzle section than Membrane 3. Table 3 shows that even with 3000 holes (Membranes 3 and 4), the TOR is increased by reducing the length of the nozzle section compared to Membrane 1, which has 6,000 holes. Comparing Membranes 3 and 4 to Membrane 2 further shows that the TOR of the nebulizer is increased by the higher number of holes (6,000 compared to 3,000) and the reduced length of the nozzle section.

[0134] In one embodiment, it is advantageous to use a laser drilling process rather than electroforming to produce the through-holes. The through-holes produced by laser drilling, as shown in Figures 5 and 6, are substantially cylindrical or conical in shape, compared to the funnel-shaped inlet and outlet of the electroformed through-holes, as disclosed in WO01 / 18280, for example. When the through-holes are substantially cylindrical or conical in shape, compared to the funnel-shaped inlet and outlet of the electroformed through-holes, the vibration of the membrane, i.e. its vibration velocity, can be transferred to the pharmaceutical formulation over a larger area by utilizing friction. The pharmaceutical formulation is then ejected from the outlet opening of the through-hole due to its own inertia, resulting in a liquid jet that breaks up to form an aerosol. Without wishing to be bound by theory, it is believed that the electroformed membrane includes a significantly curved through-hole surface, thus reducing the surface or area for energy transfer from the membrane to the liquid.

[0135] However, the invention may also be realized in electroformed membranes, where the nozzle portion is defined by a continuous portion of the through hole that begins at the smallest diameter of the through hole in the extension direction and continues toward the first surface until it reaches a diameter that is 2x or 3x the smallest diameter of the hole. In one embodiment, the entire length of the through hole is measured from the smallest diameter to the first surface.

[0136] Referring again to FIG. 1, the mouthpiece 3 has an opening 6 sealed by an elastic valve element 7 (exhalation valve) so that the patient does not have to remove the therapeutic device or take it down from his / her mouth after inhaling the aerosol. When the patient exhales into the mouthpiece 3 and thus into the nebulization chamber 2, the elastic valve element 7 opens to allow exhaled air to escape from the interior of the therapeutic aerosol. When inhaling, ambient air flows through the nebulization chamber 2. The nebulization 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 inhales from the nebulization chamber 2, the elastic valve element opens to allow ambient air to enter the nebulization chamber to mix with the aerosol and remain inside the nebulization chamber 2 to be inhaled. Further description of this process is provided in U.S. Pat. No. 6,962,151, which is incorporated herein by reference in its entirety for all purposes.

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

[0138] On the face facing the end wall 12, a cylindrical liquid container 10 is open. The opening is used to pour liquid into the liquid reservoir 10. A short distance 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 a suitably realized opening in the housing 35.

[0139] The open end of the liquid container 10 is closed by a flexible sealing element 16. The sealing element 16 is at the end of the peripheral wall 14 of the liquid container 10 and extends into the interior of the liquid container 10 in a pot-like manner, forming a conically extending wall section 17 of the sealing element 16, which is closed by a flat wall section 18 of the sealing element 16. As discussed further below, forces act through this flat wall section 18 on the sealing element 16, so that in one embodiment the flat wall section 18 is thicker than the other sections of the sealing element 16. There is a distance to the conical wall section 17 at the periphery of the flat wall section 18 such that the conical wall section 17 can fold when the flat wall section 18 is moved upwards compared to the representation in FIG. 2.

[0140] The face of the flat wall section 18 facing away from the interior of the liquid container has a protrusion that includes a frusto-conical section 19 and a cylindrical section 20. As the flexible material of the sealing element 16 allows the deformation of the frusto-conical section 19, this design allows the protrusion to be introduced into a matching opening of the cylindrical section and latched.

[0141] In one embodiment, the aerosol generator 4 comprises a slidable sleeve 21 with this type of opening, which is essentially an open-sided hollow cylinder. The opening for mounting the sealing element 16 is realized in the end wall of the slidable sleeve 21. With the truncated cone 19 latched in place, the end wall of the slidable sleeve 21, including the opening, rests on the flat sealing element wall section 18. The latching of the truncated cone 19 to the slidable sleeve allows the transmission of forces from the slidable sleeve 21 to the flat wall section 18 of the sealing element 16, such that the sealing section 18 follows the movements of the slidable sleeve 21 in the central longitudinal axis of the liquid container 10.

[0142] In a generalized form, the slidable sleeve 21 can be seen as a slidable element, which can also be realized as a slidable rod that can be fitted or inserted into a drilled hole, for example. A feature of the slidable element 21 is the fact that it can be used to apply a substantially linear force on the flat wall element 18 of the sealing element 16. Overall, the decisive factor for the mode of operation of the aerosol generator of the invention is the fact that the slidable element transmits a linear movement to the sealing element, such that an increase in volume occurs in the reservoir 10. This causes the generation of a negative pressure in the reservoir 10, since the latter is otherwise airtight.

[0143] The sealing element 16 and the slidable element 21 can be manufactured in one piece, i.e. in one step, but from different materials. Manufacturing techniques for this can be utilized such that the integral components of the nebulizer are produced, for example, in a fully automated manufacturing step.

[0144] In one embodiment, the slidable sleeve 21 is open at the end facing the drilled hole for the truncated cone, but at least two diametrically opposed lugs 22 and 23 project radially into the interior of the slidable sleeve 21. A collar 24 surrounds the slidable sleeve and extends radially outward. The collar 24 serves as a support for the slidable sleeve 21 in the position shown in FIG. 5, while the lugs 22 and 23 projecting into the interior of the slidable sleeve 21 are used to absorb forces acting on the slidable sleeve 21, particularly parallel to the central longitudinal axis. In one embodiment, these forces are generated by means of 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 one of the protrusions 22 or 23 and one groove 25. In a further embodiment, two or more protrusions arranged in an evenly distributed manner and a corresponding number of grooves are provided.

[0146] In one embodiment, the rotating sleeve 26 is also an open-sided cylinder, with its open end disposed within the slidable sleeve 21 facing the truncated cone 19 and allowing the truncated cone 19 to enter the rotating sleeve 26. In addition, the rotating sleeve 26 is disposed within the slidable sleeve 21 in such a way that the projections 22 and 23 are within the helical groove 25. The inclination of the helical groove 25 is designed such that, when the rotating sleeve 26 is rotated relative to the slidable sleeve 21, the projections 22 and 23 slide along the helical groove 25, exerting a force parallel to the central longitudinal axis on the sliding projections 22 and 23 and thus on the slidable sleeve 21. This force displaces the slidable sleeve 21 in the direction of the central longitudinal axis, so that the sealing element 16, which is latched into the drilled hole of the slidable sleeve by means of the truncated cone, 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 creates a negative pressure in the liquid container 10 that depends, among other things, on the distance that the slidable sleeve 21 is displaced in the direction of the central longitudinal axis. This displacement reduces the initial volume V of the gas-tight liquid container 10. RI Let the volume be V RN 2. This displacement is then defined by the design of the spiral groove 25 in the rotating sleeve 26. Thus, the aerosol generator of the present invention ensures that a negative pressure in the reservoir 10 can be created in the relevant area using simple structural means.

[0148] To ensure that the force applied to create the negative pressure when handling the device is kept low, the rotating sleeve 26 is realized integrally with a handle 27 having a size selected to allow the user to effortlessly rotate the handle 27, and thus the rotating sleeve 26, by hand. The handle 27 has substantially the shape of an open-sided flattened cylinder or truncated cone such that at its outer edge a peripheral gripping area 28 is formed which the user's hand contacts in order to turn the handle 27.

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

[0150] In order to create a unit that can be easily and uniformly operated from the slidable sleeve 21 and the rotating sleeve 26 including the handle 27, in one embodiment the aerosol generator described herein has a bearing sleeve 29 for receiving the slidable sleeve 21, which essentially comprises a flat cylinder with an open side. The diameter of the peripheral wall 30 of the bearing sleeve 29 is smaller than the inner diameter of the handle 27 and, in the example of the embodiment described, is adapted to the inner diameter of a cylindrical latching ring 31, which is arranged concentrically with the gripping area 28 of the handle 27, but has a smaller diameter, on the face of the handle 27 on which the rotating sleeve 26 is also located. On the face of the cylindrical latching ring 31 facing the rotating sleeve, peripheral latching edges 32 are realized which can be engaged with latching lugs 33 spaced apart on the peripheral wall 30 of the bearing sleeve 29. This allows the handle 27 to be placed onto the bearing sleeve 29 so that it rests over the open end of the bearing sleeve 29 and the latching edge 32 interlatches with the latching lug 33, as shown in Figure 5.

[0151] To hold the slidable sleeve 21, an opening is provided in the center of the sealed end of the bearing sleeve 29, in which the slidable sleeve 21 is placed, as can be seen in Figure 2. The collar 24 of the slidable sleeve 21 is located on the surface of the end wall of the bearing sleeve 29 facing the handle, in the position shown in Figure 2. Extending into the bearing opening are two diametrically opposed projections 51 and 52, which project into two longitudinal grooves 53 and 54 on the peripheral surface of the slidable sleeve 21. The longitudinal grooves 53 and 54 run parallel to the longitudinal axis of the slidable sleeve 21. The guide projections 51 and 52 and the longitudinal grooves 53 and 54 provide an anti-rotation locking for the slidable sleeve 21, so that a rotational movement of the rotating sleeve 26 does not result in a rotation of the slidable sleeve 21, but in a linear displacement. 2, this ensures that the slidable sleeve 21 is held in the assembly of the handle 27 and the bearing sleeve 29 in an axially displaceable but rotationally locked manner. When the handle 27 is rotated relative to the bearing sleeve 29, the rotating sleeve 26 also rotates relative to the slidable sleeve 21, causing the sliding lugs 22 and 23 to move along the helical groove 25. This causes the slidable sleeve 21 to be axially displaced in the opening of the bearing sleeve 29.

[0152] It is also possible to dispense with the guide projections 51 and 52 in the bearing opening and the longitudinal grooves 53 and 54 in the slidable 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 the truncated cone 19, the cylindrical section 20 of the sealing element 16 and the large-area support for the slidable sleeve 21 holding the truncated cone on the flat sealing element section 18 achieve anti-rotational locking of the slidable sleeve 21 by means of friction. In a further embodiment, the sealing element 16 is fixed so that it cannot rotate relative to the bearing sleeve 29.

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

[0154] In one embodiment, the necessary force is generated, at least in part, by the interaction between the handle 27 and the housing 35, in which the pharmaceutical agent reservoir is either integrally realised or in which the pharmaceutical agent (liquid) reservoir 10 is inserted as shown in Figure 2. In this case, the pharmaceutical agent reservoir 10, inserted into the casing by means of the marginal projections 15, rests at intervals on supports 36 in the housing 35 extending 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, the supports are only provided at intervals, so that openings for ambient air are provided when the patient inhales, as will be explained in more detail below.

[0155] In figure 2 one can see the rotation lock, which is realised on the one hand by means of the handle 27 and on the other hand by means of the housing 35. The locking lugs 62 and 63 on the housing 35 are shown. However, in so far as the device of the invention is concerned with the creation of a negative pressure in the reservoir 10, there are no special requirements as regards the design of the rotation lock.

[0156] In one embodiment, 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, such that when the liquid reservoir 10 contains (is filled or injected with) an 8 mL amount of liquid (e.g., an aminoglycoside pharmaceutical formulation) to be released in the form of an aerosol, an air cushion of 8 mL or about 8 mL is obtained. RN That is, the initial volume V of the liquid in the liquid reservoir 10 L for volume V RN The ratio of is at least 2.0 and the volume of gas V A and the volume of the liquid V L The ratio between volumes V of about 15.5 mL, about 19.5 mL and about 22.5 mL is at least 1.0. RN The reservoir with V RN It has been shown that the β-stimulus γ-stimulus increases with increasing β-stimulus.

[0157] In one embodiment, 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. 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. 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 about 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 about 6 mL to about 15 mL, and V RN and V L In a further embodiment, the ratio between V RN and V L is at least about 2.0 to at least about 2.8.

[0159] The volume of the air cushion, in one embodiment, 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 initial volume of liquid V L for volume V RN Theoretically, the ratio of the increased volume V of the liquid reservoir 10 is at least 2.0. RN An infinite expansion of V would result in a nearly stable negative pressure range. L for volume V RN The ratio of V to V is in the range of 2.0 to 4.0, and in a further embodiment is 2.4 to 3.2. L Regarding the ratio range (V RN / V L ) are provided in Table 4 below. [Table 4]

[0161] The systems provided herein can be used to treat a variety of pulmonary infections in subjects in need of such treatment. Pulmonary infections (e.g., in cystic fibrosis patients) that can be treated with the methods of the present invention include gram-negative infections. In one embodiment, infections caused by the following bacteria are treatable with the systems and formulations provided herein: Pseudomonas (e.g., P. aeruginosa, P. paucimobilis, P. putida, P. fluorescens, and P. acidovorans), Burkholderia (e.g., B. pseudomallei, B. cepacia, B. cepacia complex, B. dolosa, B. fungorum, B. gladioli, B. multivorans, B. vietnamiensis, B. pseudomallei, B. 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), Methicillin-resistant Staphylococcus aureus (MRSA), Streptococcus (e.g. Streptococcus pneumoniae), Escherichia coli, Klebsiella, Enterobacter, Serratia, Haemophilus, Yersinia pestis, Mycobacterium, nontuberculous mycobacteria (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. conpicuum, 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 infections caused by nontuberculous mycobacterial infections. In one embodiment, the system described herein is used to treat infections 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 Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium, or Mycobacterium avium complex infections. In yet another embodiment, the Mycobacterium avium infection is Mycobacterium avium subsp.hominissuis.

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

[0164] In one embodiment, the system provided herein is used to treat or prevent Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium, or Mycobacterium avium complex pulmonary infections in cystic fibrosis or non-cystic fibrosis patients. 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, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamycin, or combinations thereof. In yet another embodiment, the aminoglycoside is amikacin, e.g., amikacin sulfate.

[0165] An obstacle to the treatment of infectious diseases such as Pseudomonas aeruginosa, a major cause of chronic disease in cystic fibrosis patients, is drug penetration into the sputum / biofilm barrier on epithelial cells (Figure 7). In Figure 7, the doughnut shapes represent liposomal / complexed aminoglycosides, the "+" symbols represent free aminoglycosides, the "-" symbols represent mucin, alginate and DNA, and the solid bar symbol represents Pseudomonas aeruginosa. This barrier contains both colonized and planktonic P. aeruginosa embedded in alginate or exopolysaccharides from the bacteria, both of which have a net negative charge, as well as DNA from damaged leukocytes and mucin from lung epithelial cells. Negative charges bind positively charged drugs such as aminoglycosides, preventing their penetration and rendering them biologically ineffective (Mendelman et al., 1985). Without wishing to be bound by theory, encapsulation of aminoglycosides in liposomes or lipid complexes shields or partially shields the aminoglycoside from non-specific binding to sputum / biofilms, allowing penetration of the liposomes or lipid complexes in which the aminoglycoside is encapsulated (Figure 7).

[0166] In another embodiment, a patient is treated with one of the systems provided herein for a nontuberculous mycobacterial pulmonary infection. In a further 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 a cystic fibrosis patient. 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 a patient 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 lung infections in non-CF bronchiectasis patients. In a further embodiment, the system provided herein comprises a liposomal aminoglycoside formulation. In a further embodiment, the aminoglycoside is amikacin.

[0170] As provided herein, the present invention provides aminoglycoside formulations administered by inhalation, in one embodiment, the MMAD of the aerosol is about 3.2 μm to about 4.2 μm as measured by an Andersen Cascade Impactor (ACI), or about 4.4 μm to about 4.9 μm as measured by a Next Generation Impactor (NGI).

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

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

[0173] The present invention will be further illustrated with reference to the following examples, although it should be noted that these examples, like the above embodiments, are illustrative and should not be construed as limiting the scope of the invention in any way. Example 1: Comparison of nebulizer reservoir volumes

[0174] In this example, the aerosol generator was an investigational eFlow® nebulizer from Pari Pharma GmbH (Germany) modified for use with the liposomal aminoglycoside formulations provided herein. The first aerosol generator had an initial volume of reservoir V RI The first was 13 mL (A), the second was 17 mL (B), the third was 22 mL (C), and the fourth was 20 mL (D). RN the first 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 liposomal amikacin formulation was poured into reservoir 10. As shown in Figure 8, the 8 mL air cushion resulted in an aerosol generation period of 14 to 16 minutes upon complete release of the 8 mL of formulation in the reservoir. However, the 12 mL air cushion reduced the aerosol generation time to a range of 12 to approximately 13 minutes. The 17 mL air cushion further reduced the aerosol generation time to a range of 10 to 12 minutes (Figure 6).

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

[0177] The dependence of aerosol generator efficiency (proportional to liquid or total discharge rate) on different negative pressures was measured with the nebulizer described above. Liposomal 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 experiments. As shown in Figure 10, the efficiency is optimal in the negative pressure range of 150 mbar to 300 mbar. As also 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 modified eFlow® based aerosol generators, where the first aerosol generator (A) had an increased volume of the reservoir V RN The modified eFlow® had a volume of 19.5 mL and was filled with 8 mL of the liposomal amikacin formulation.

[0179] The second aerosol generator (B) has an increased volume V RN a third aerosol generator (C) having a reservoir filled with 8 mL of the liposomal amikacin formulation and an increased volume V of 24.5 mL; RN and was filled with 8 mL of the above liquid. The fourth aerosol generator had an increased volume of the liquid reservoir V RN The volume of the tube was 22.5 mL and was filled with 8 mL of the above liposomal amikacin formulation.

[0180] Figure 11 shows the experimental data for these four aerosol generators loaded with 8 mL of the liposomal amikacin formulation. These results show that the aerosol generation time to completely release the liposomal amikacin formulation in the reservoir varies with the initial volume of liquid in the reservoir before use (V L ) versus the increased volume of the reservoir (V RN ) ratio. FIG. 11 shows that the modified aerosol generator device (A) required approximately 16 minutes of aerosol generation time, whereas the aerosol generation time was approximately 10 minutes for the ratio V RN / V LThe data also show that with 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 show that a larger air cushion allows the aerosol generator to operate in the efficient negative pressure range for a longer period of time, thereby significantly reducing the total aerosol generation time. Thus, even a large amount of liquid, e.g., 8 mL, can be nebulized (emitted in the form of an aerosol) in a period of less than 12 minutes. Example 2: Aerosol Properties of Amikacin Formulations

[0182] Eleven different lots of liposomal amikacin formulations were tested in a modified eFlow® nebulizer (i.e., modified for use with the liposomal aminoglycoside formulations described herein) with a modified 40 mesh membrane fabricated as described herein and a reservoir with 8 mL liquid volume and air cushion as described above. Aerosol characteristics, namely mass median aerodynamic diameter (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF), were determined by cascade impaction using either the ACI (Andersen Cascade Impactor) or NGI (Next Generation Impactor). Mass Median Aerodynamic Diameter (MMAD) Measurements by ACI

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

[0184] When the 8 mL handset was loaded with 8 mL, it was found that the ACI was unable to handle the entire 8 mL dose; i.e., the amikacin liposomal formulation deposited on ACI plate 3 overflowed. It was determined that the percentage drug distribution at each ACI stage was not affected by the amount of liposomal amikacin formulation collected inside the ACI, unless there was liquid overflow at ACI stage 3 (data not shown). Thus, for nebulization, the nebulizer was filled with 4 mL of the liposomal amikacin formulation and nebulized until empty, or filled with 8 mL of the liposomal amikacin formulation and nebulized over a collection time of approximately 6 minutes (i.e., approximately 4 mL).

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

[0186] After collecting the nebulization, ACI collection plates 0, 1, 2, 3, 4, 5, 6, and 7 were removed and each was placed on its own Petri dish. 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. Samples from plates 0, 1, 2, 3, 4, 5, and 6 were further diluted appropriately with mobile phase C for HPLC analysis. Samples from plate 7 were directly analyzed by HPLC without further dilution. The ACI filter was also transferred to a 20 mL vial, 10 mL of extraction solution was added, and the capped vial was vortexed to dissolve any formulation deposited thereon. The liquid samples from the vials were filtered (0.2 μm) into HPLC vials for HPLC analysis. The inlet port and connector were also rinsed with 10 mL of extraction solution to dissolve the formulation deposited therein, and samples were collected and subjected to HPLC analysis at a dilution of 2. The mass median aerodynamic diameter (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF) were calculated based on the amount of amikacin deposited at each stage of the impactor.

[0187] For nebulizers loaded with 8 mL and nebulized for 6 minutes, fine particle dose (FPD) was normalized to the volume of nebulized formulation to compare FPD across all experiments. FPD (normalized to the volume of nebulized formulation) was calculated according to the following equation:

number

[0188] A Next Generation Impactor (NGI) was also used for MMAD measurements, and the nebulization procedure was performed inside a ClimateZone chamber (Westech Instruments Inc., GA) to maintain temperature and RH% during nebulization. The ClimateZone was preset to a temperature of 18°C ​​and a relative humidity of 50%. The NGI was assembled and loaded inside the ClimateZone. A probe thermometer (VWR Dual Thermometer) was attached to the surface of the NGI to monitor the temperature of the NGI. Nebulization was initiated when the temperature of the NGI reached 18 ± 0.5°C.

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

[0190] After aerosol collection, the NGI tray was removed from the NGI along with the tray holder. Appropriate amount of extraction solution was added to NGI cups 1, 2, 3, 4, 5, 6, 7 and MOC to dissolve the formulation deposited in these cups. This material was transferred to a volumetric flask respectively. For NGI cups 1, 2, and 6, a 25 ml volumetric flask was used, and for NIG cups 2, 3, and 4, a 50 ml volumetric flask was used. More extraction 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 cup to the volumetric flask. The volumetric flask was topped off to a final volume of 25 ml or 50 ml and shaken well before sampling. 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 directly 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 any formulation deposited thereon. The liquid sample from the vial was filtered (0.2 micron) into an HPLC vial for HPLC analysis. The inlet port and connector were also rinsed with 10 mL of extraction solution to dissolve any formulation deposited thereon, and a sample was collected and subjected to HPLC analysis at an 11-fold dilution.

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

[0192] To compare FPD across all experiments, FPD was normalized to the volume of nebulized formulation. FPD (normalized to the volume of nebulized formulation) was calculated according to the following equation:

number

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

[0194] Nebulization rate studies (grams of formulation nebulized per minute) were performed 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 volume of formulation was added and the nebulizer device was weighed again (W2). The nebulizer and timer were started and the nebulized formulation was collected in a cooled impinger at a flow rate of approximately 8 L / min (see FIG. 14 for details of the experimental setup). When no aerosol was anymore observed, the timer was stopped. The nebulizer was weighed again (W3) and the time of nebulization (t) was recorded. The total amount of nebulized formulation was calculated as W2-W3, and the total drug remaining after nebulization was calculated as W3-W1. The nebulization rate of the formulation was calculated according to the following formula:

number

[0195] The nebulization rates (in g / min) and other relevant results for liposomal amikacin nebulized using the nebulizers constructed according to the present specification (24 aerosol heads were selected and used in these studies) are recorded in Table 6. [Table 6-1] [Table 6-2] [Table 6-3] Example 4: Percentage of associated amikacin after nebulization and characterization of the nebulizer

[0196] Free and liposome-complexed amikacin was measured in the nebulizer of Example 3. As described in Example 3, the nebulizer was collected in a chilled impinger (FIG. 14) at a flow rate of 8 L / min.

[0197] The nebulized material collected in the impinger was rinsed with 1.5% NaCl and transferred to a 100 mL or 50 mL volumetric flask. The impinger was then rinsed several times with 1.5% NaCl to transfer all of the formulation deposited on the impinger to the flask. To measure the free amikacin concentration of the nebulized material, 0.5 mL of the diluted nebulized material inside the volumetric flask was removed and centrifuged with an Amicon® Ultra-0.5 mL 30K centrifuge filter. The aerosol was placed in a volumetric flask (regenerated cellulose, 30K MWCO, Millipore) and the apparatus was centrifuged at 5000G for 15 minutes at 15°C. An appropriate amount of the filtrate was taken and diluted 51-fold with mobile phase C solution. The amikacin concentration was determined by HPLC. To measure the total amikacin concentration of the aerosol, an appropriate amount of the diluted aerosol inside the volumetric flask was taken and diluted (and dissolved) 101-fold in 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 amikacin associated after nebulization was calculated according to the following formula:

number

[0199] The percentage of amikacin associated after nebulization and the total dose recovery from the nebulization experiments described in Table 6 are summarized in Table 7. The corresponding nebulization rates are also included in Table 7. [Table 7-1] [Table 7-2] [Table 7-3]

[0200] The total concentration of amikacin in the liposomal amikacin formulation was measured for the remaining samples during the study using the same HPLC and amikacin standard. The value obtained was 64 mg / mL amikacin. The % associated amikacin values ​​after nebulization ranged from 58.1% to 72.7%, with a mean value of 65.5 ± 2.6%. When 8 mL of the liposomal amikacin formulation was nebulized, the total recovery of amikacin ranged from 426 mg to 519 mg, with a mean value of 476 ± 17 mg. The calculated amount of amikacin nebulized (by weight of the nebulized liposomal amikacin formulation in Table 7) ranged from 471 mg to 501 mg, with a mean value of 490 ± 8 mg. The total amikacin recovery ranged from 91% to 104%, with a mean value of 97 ± 3% (n = 72). Liposome size

[0201] Pre- and post-nebulized liposomal amikacin formulations (64 mg / mL amikacin) were appropriately diluted with 1.5% NaCl, and liposome particle size was measured by light scattering using a Nicomp380 Submicron Particle Sizer (Nicomp, Santa Barbara, Calif.).

[0202] Liposome size was measured after nebulization of liposomal amikacin formulations aerosolized using 24 nebulizer aerosol heads with an 8 mL reservoir handset. Liposome sizes ranged from 248.9 nm to 288.6 nm with a mean of 264.8 ± 6.7 nm (n = 72). These results are presented in Table 8. The mean liposome diameter before nebulization was approximately 285 nm (284.5 nm ± 6.3 nm). [Table 8-1] [Table 8-2] [Table 8-3]

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

[0204] The embodiments illustrated and described herein are intended merely to teach those skilled in the art the best way known to the inventors to make and use the invention. As those skilled in the art will recognize in light of the above teachings, modifications and variations can be made to the above-described embodiments of the invention without departing from the invention. It is therefore to be understood that, within the scope of the appended claims and their equivalents, the invention may be practiced otherwise than as specifically described.

Claims

[Claim 1] A pharmaceutical formulation as described in the specification.

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