HK1262634B - Systems for treating pulmonary infections - Google Patents

Description

System for treating lung infections

This application is a patent application filed in China with application number 201380030763.4, entitled “System for treating lung infection” and filed on May 21, 2013 (PCT application number

PCT/US2013/042113).

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority to U.S. Provisional Application No. 61/649,830, filed May 21, 2012, which is hereby incorporated by reference in its entirety.

Background of the Invention

Certain technologies suitable for inhaled drug delivery employ liposomes and lipid complexes to provide prolonged drug therapeutic effects in the lungs. These technologies also provide drugs with sustained activity, as well as the ability to target and enhance drug uptake to disease sites.

Inhalation delivery of liposomes is complicated by their sensitivity to shear-induced stress during nebulization, which can lead to changes in physical properties (e.g., entrapment, size). However, this need not prohibit their use in drug development as long as the changes in properties are reproducible and meet acceptance criteria.

Cystic fibrosis (CF) patients have thick mucus and/or saliva secretions in their lungs, frequent major infections, and biofilms caused by bacterial colonization. All of these fluids and materials create obstacles for aminoglycosides to effectively target infection. Liposomal aminoglycoside formulations can be used to combat bacterial biofilms.

SUMMARY OF THE INVENTION

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

In one aspect, the present invention provides a system for treating or providing prevention of lung infection. In one embodiment, the system includes a pharmaceutical preparation and a nebulizer, wherein the pharmaceutical preparation comprises a liposome-complexed aminoglycoside, wherein the preparation is a dispersion (e.g., a liposomal solution or suspension), the lipid component of the liposome is composed of electrically neutral lipids, and the nebulizer generates an aerosol of the pharmaceutical preparation at a rate greater than about 0.53 g per minute. In one embodiment, the total aerodynamic median diameter (mass median aerodynamic diameter, MMAD) of the aerosol is measured by an Anderson Cascade Impactor (ACI) to be less than about 4.2 μm; measured by ACI to be about 3.2 μm to about 4.2 μm; or measured by a next generation impactor (NGI) to be less than about 4.9 μm; or measured by NGI to be about 4.4 μm to about 4.9 μm.

In another embodiment, a system for treating or providing prevention of a pulmonary infection comprises a pharmaceutical formulation comprising a liposomally complexed aminoglycoside and a nebulizer, wherein the formulation is a dispersion (e.g., a liposomal solution or suspension), the lipid component of the liposomes is composed of electrically neutral lipids, and the nebulizer generates an aerosol of the pharmaceutical formulation at a rate of greater than about 0.53 g per minute, and the fine particle fraction (FPF) of the aerosol is greater than or equal to about 64% as measured by an Anderson Cascade Impactor (ACI), or greater than or equal to about 51% as measured by a New Generation Impactor (NGI).

In one embodiment, the application provides a system including a pharmaceutical preparation comprising an aminoglycoside. In a further embodiment, the aminoglycoside is amikacin, apramycin, arbekacin, astromicin, capreomycin, dibekacin, framycetin, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, verdamicin, or a combination thereof. In yet a further embodiment, the aminoglycoside is amikacin. In another embodiment, the aminoglycoside is selected from the aminoglycosides listed in Table A below, or a combination thereof.

The pharmaceutical preparations provided herein are liposome dispersions (i.e., liposome dispersions or aqueous liposome dispersions, which can be liposome solutions or liposome suspensions). In one embodiment, the lipid component of the liposome is essentially composed of one or more electrically neutral lipids. In a further embodiment, the electrically neutral lipid comprises a phospholipid and a sterol. In a further embodiment, the phospholipid is dipalmitoylphosphatidylcholine (DPPC) and the sterol is cholesterol.

In one embodiment, the ratio of lipid to drug in the aminoglycoside drug 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.

In one embodiment, after nebulization, the aerosolized aminoglycoside formulation 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 a further embodiment, the amikacin is amikacin sulfate.

In one embodiment, prior to nebulization, about 70% to about 100% of the aminoglycoside present in the formulation is liposomally complexed, for example, encapsulated in a plurality of liposomes. In a further embodiment, the aminoglycoside is selected from the aminoglycosides provided in Table A. In a further embodiment, the aminoglycoside is amikacin. In yet a further embodiment, about 80% to about 100% of the amikacin is liposomally complexed, or about 80% to about 100% of the amikacin is encapsulated in a plurality of liposomes. In another embodiment, prior to nebulization, about 80% to about 100%, about 80% to about 99%, about 90% to about 100%, 90% to about 99%, or about 95% to about 99% of the aminoglycoside present in the formulation is liposomally complexed prior to nebulization.

In one embodiment, after nebulization, the percentage of liposome-complexed (also referred to herein as "liposome-bound") aminoglycoside 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 provided in Table A. In a further embodiment, the aminoglycoside is amikacin. In yet a further embodiment, the amikacin is amikacin sulfate.

On the other hand, the present invention provides treatment or provides the method for preventing lung infection.In one embodiment, lung infection is the lung infection (this application is also referred to as Gram-negative bacteria infection) caused by Gram-negative bacteria.In one embodiment, lung infection is Pseudomonas (Pseudomonas) infection, for example, Pseudomonas aeruginosa (Pseudomonas aeruginosa) infection.In another embodiment, lung infection is caused by the one in the Pseudomonas species provided in Table B below.In one embodiment, the mycobacterium lung infection of a kind of system treatment patient provided by the application.In a further embodiment, mycobacterium lung infection is nontuberculous mycobacterium lung infection, mycobacterium abscessus lung infection or mycobacterium avium complex lung infection.In one or more of the aforementioned embodiments, the patient is a cystic fibrosis patient.

In one embodiment, a system provided herein is used to treat a lung infection in a cystic fibrosis patient. In a further embodiment, the lung infection is caused by Mycobacterium abscessus, Mycobacterium avium complex, or Pseudomonas aeruginosa. In another embodiment, the lung infection is caused by a species selected from the group consisting of Mycobacterium avium, Mycobacterium avium hominis subsp. suis (MAH), Mycobacterium abscessus, Mycobacterium chelonae, Mycobacterium bovis, Mycobacterium kansasii, Mycobacterium ulcerans, Mycobacterium avium, Mycobacterium avium complex (MAC) (Mycobacterium avium and Mycobacterium intracellulare), Mycobacterium spp., Mycobacterium kansasii, Mycobacterium exogenum, Mycobacterium immunogenicum, Mycobacterium toad, Mycobacterium marinum, Mycobacterium malmoe, Mycobacterium marinum, Mycobacterium mucogenes The disease is caused by nontuberculous mycobacteria such as Mycobacterium simplicis, Mycobacterium smegmatis, Mycobacterium surga, Mycobacterium terrestris, Mycobacterium terrestris complex, Mycobacterium hemophilus, Mycobacterium geneva, Mycobacterium asiatica, Mycobacterium subspecies, Mycobacterium gordonii, Mycobacterium nonchromogenic, Mycobacterium tristis, Mycobacterium lentiflavum, Mycobacterium occulta, Mycobacterium fortuitum, Mycobacterium fortuitum complex (Mycobacterium fortuitum and Mycobacterium chelonae), or a combination thereof.

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

In one embodiment, the method comprises aerosolizing a pharmaceutical formulation comprising a liposomally complexed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate greater than about 0.53 grams per minute. The method further comprises administering the aerosolized pharmaceutical formulation to the patient's lungs; wherein the aerosolized pharmaceutical formulation comprises a mixture of free aminoglycoside and liposomally complexed aminoglycoside (e.g., aminoglycoside encapsulated in liposomes), and the liposome component of the formulation consists of electrically neutral lipids. In yet further embodiments, 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.

In another aspect, a liposomally complexed aminoglycoside aerosol (e.g., a liposomally 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 liposomally complexed and the aerosol is generated at a rate of greater than about 0.53 grams per minute. In a further embodiment, about 65% to about 75% of the aminoglycoside is liposomally complexed and the aerosol is generated at a rate of greater than about 0.53 grams per minute. In any of the foregoing embodiments, the aerosol is generated at a rate of greater than about 0.54 grams per minute. In any of the foregoing embodiments, the aerosol is generated at a rate of greater than about 0.55 grams per minute. In any of the foregoing embodiments, the aminoglycoside is selected from the aminoglycosides provided in Table A.

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

In one embodiment, the FPF of the lipid-complexed aminoglycoside aerosol is measured by an Anderson cascade impactor (ACI) as greater than or equal to about 64%, or by a new generation impactor (NGI) as greater than or equal to about 51%. In a further embodiment, the aerosol comprises an aminoglycoside and a plurality of liposomes, the liposomes comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposome-complexed, for example, 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 of the foregoing embodiments, the aerosol is generated at a rate greater than about 0.54 grams per minute. In any of the foregoing embodiments, the aerosol is generated at a rate greater than about 0.55 grams per minute. In any of the foregoing embodiments, the aminoglycoside is selected from the aminoglycosides provided in Table A.

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).

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, from about 70 mg/mL to about 75 mg/mL. In a further embodiment, the aminoglycoside is selected from the aminoglycosides provided in Table A. In yet a further embodiment, the aminoglycoside is amikacin (e.g., amikacin sulfate).

This application also relates to the following:

1. A system for treating or providing prophylaxis for a lung infection in a patient, comprising:

(a) a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein the formulation is an aqueous dispersion and the lipid component of the liposomes consists of electrically neutral lipids, and

(b) a nebulizer that generates an aerosol of the pharmaceutical formulation at a rate of greater than about 0.53 g per minute, wherein the aerosol has a total median aerodynamic diameter (MMAD) of less than about 4.2 μm as measured by an Anderson Cascade Impactor (ACI) or less than about 4.9 μm as measured by a New Generation Impactor (NGI).

2. A system for treating or providing prevention of lung infection, comprising:

(a) a pharmaceutical formulation comprising a liposome-complexed aminoglycoside, wherein the formulation is an aqueous dispersion and the lipid component of the liposomes consists of electrically neutral lipids, and

(b) a nebulizer that generates an aerosol of the pharmaceutical formulation at a rate of greater than about 0.53 g per minute, wherein the aerosol has a fine particle fraction (FPF) of greater than or equal to about 64% as measured by ACI, or greater than or equal to about 51% as measured by NGI.

3. The system of claim 1 or 2, wherein the aminoglycoside is selected from amikacin, apramycin, arbekacin, astamicin, capreomycin, dibekacin, neomycin B, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribosomycin, sisomicin, spectinomycin, streptomycin, tobramycin, methylthiazolinone, or a combination thereof.

4. The system of any one of items 1 to 3, wherein the aminoglycoside is amikacin.

5. The system of any one of items 1 to 4, wherein the aminoglycoside is amikacin sulfate.

6. The system of any one of items 1 to 5, wherein the liposomes comprise unilamellar vesicles, multilamellar vesicles, or a mixture thereof.

7. The system of any one of items 1 to 6, wherein the neutral lipid comprises a neutral phospholipid or comprises a neutral phospholipid and a sterol.

8. The system of any one of items 1 to 7, wherein the electrically neutral lipid comprises phosphatidylcholine and a sterol.

9. The system of any one of items 1 to 8, wherein the neutral lipid comprises dipalmitoylphosphatidylcholine (DPPC) and a sterol.

10. The system of any one of items 1 to 9, wherein the electroneutral lipid comprises DPPC and cholesterol.

11. The system of any one of items 1 to 10, wherein the aminoglycoside is amikacin, the electrically neutral lipids consist of DPPC and cholesterol, and the liposomes comprise unilamellar vesicles, multilamellar vesicles, or a mixture thereof.

12. The system of any one of items 1 to 11, wherein the weight ratio of free aminoglycoside to liposome-complexed aminoglycoside is from about 1:100 to about 100:1.

13. The system of any one of items 1 to 12, wherein the weight ratio of free aminoglycoside to the liposome-complexed aminoglycoside is from about 1:10 to about 10:1.

14. The system of any one of items 1 to 13, wherein the weight ratio of free aminoglycoside to liposome-complexed aminoglycoside is from about 0.3:1 to about 2:1.

15. The system of any one of items 1 to 14, wherein the volume of the drug formulation is about 8 mL.

16. The system of any one of items 1 to 15, wherein the aerosol comprises about 55% to about 75% liposomally complexed amikacin.

17. The system of any one of items 1 to 16, wherein the MMAD of the liposomal complexed aminoglycoside is about 3.2 μm to about 4.2 μm as measured by ACI; or about 4.4 μm to about 4.9 μm as measured by NGI.

18. The system of any one of items 1 to 17, wherein the MMAD of the liposomal complexed aminoglycoside is 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.

19. The system of any one of items 1 to 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.

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.

21. The system of any one of items 1 to 20, wherein the nebulizer generates an aerosol of the pharmaceutical formulation at a rate greater than about 0.54 g per minute.

22. The system of any one of items 1 to 21, wherein the aerosol comprises an amount of free aminoglycoside effective to provide immediate bactericidal or antibacterial activity against pulmonary infections and an amount of liposomal-complexed aminoglycoside effective to provide sustained bactericidal or antibacterial activity against pulmonary infections.

23. The system of any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 500 mg to about 650 mg of the aminoglycoside.

24. The system of any one of items 1 to 22, wherein the pharmaceutical formulation comprises from about 550 mg to about 625 mg of an aminoglycoside.

25. The system of any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 550 mg to about 600 mg of an aminoglycoside.

26. The system of any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 560 mg of an aminoglycoside.

27. The system of any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 580 mg of an aminoglycoside.

28. The system of any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 590 mg of an aminoglycoside.

29. The system of any one of items 1 to 22, wherein the pharmaceutical formulation comprises about 600 mg of an aminoglycoside.

30. A method for treating or preventing a lung infection in a patient, the method comprising:

aerosolizing a pharmaceutical formulation comprising a liposomally complexed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate greater than about 0.53 grams per minute, and

administering aerosolized drug preparations to the patient's lungs;

wherein the aerosolized pharmaceutical preparation comprises a mixture of free aminoglycoside and liposome-complexed aminoglycoside, and

The lipid components of the liposomes are composed of electrically neutral lipids, and

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.

31. The method of claim 30, wherein

The MMAD of the aerosol is about 3.2 μm to about 4.2 μm as measured by ACI, or about 4.4 μm to about 4.9 μm as measured by NGI.

32. A method of treating or providing prophylaxis for a lung infection in a patient, the method comprising:

aerosolizing a pharmaceutical formulation comprising a liposomally complexed aminoglycoside, wherein the pharmaceutical formulation is an aqueous dispersion and is aerosolized at a rate greater than about 0.53 grams per minute, and

administering the aerosolized drug formulation to the lungs of a patient;

wherein the aerosolized pharmaceutical preparation comprises a mixture of free aminoglycoside and liposome-complexed aminoglycoside,

wherein the lipid component of the liposome consists of electrically neutral lipids, and

The aerosol has an FPF of greater than or equal to about 64% as measured by ACI, or greater than or equal to about 51% as measured by NGI.

33. The method of any one of items 30-32, wherein the drug formulation is aerosolized at a rate greater than about 0.55 grams per minute.

34. The method of any one of items 30-33, wherein the drug formulation is aerosolized at a rate greater than about 0.56 grams per minute.

35. The method of any one of items 30-34, wherein the drug formulation is aerosolized at a rate greater than about 0.58 grams per minute.

36. The method of any one of items 30-35, wherein the drug formulation is aerosolized at a rate of about 0.60 to about 0.80 grams per minute.

37. The method of any one of items 30-36, wherein the drug formulation is aerosolized at a rate of about 0.60 to about 0.70 grams per minute.

38. The method of any one of items 30-37, 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.

39. The method of any one of items 30-38, wherein the pharmaceutical formulation has a volume of about 8 mL.

40. The method of any one of items 30-39, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astamicin, kanamycin B, polmycin, tobramycin, capreomycin, dibekacin, datimicin, ethimicin, neomycin B, gentamicin, H107, hygromycin, hygromycin B, myosin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomicin, ribosomycin, sisomicin, rhodestreptomycin, sorbitol, spectinomycin, saccharopolysporin, streptomycin, tobramycin, methylthiazolin, vetimicin, or a combination thereof.

41. The method of any one of items 30-40, wherein the aerosolized pharmaceutical formulation is administered once daily within a single dosing period.

42. The method of any one of items 30-41, wherein the aminoglycoside is amikacin.

43. The method of any one of items 30-42, wherein the aminoglycoside is amikacin sulfate.

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 antibacterial activity against nontuberculous mycobacterial infection, and the liposomal-complexed aminoglycoside is in an amount effective to provide sustained bactericidal or antibacterial activity against nontuberculous mycobacterial infection.

45. The method of any one of items 30-44, wherein the pharmaceutical formulation comprises from about 500 mg to about 650 mg of the aminoglycoside.

46. The method of any one of items 30-44, wherein the pharmaceutical formulation comprises from about 550 mg to about 625 mg of the aminoglycoside.

47. The method of any one of items 30-44, wherein the pharmaceutical formulation comprises from about 550 mg to about 600 mg of the aminoglycoside.

48. The method of any one of items 30-44, wherein the pharmaceutical formulation comprises about 560 mg of an aminoglycoside.

49. The method of any one of items 30-44, wherein the pharmaceutical formulation comprises about 580 mg of an aminoglycoside.

50. The method of any one of items 30-44, wherein the pharmaceutical formulation comprises about 590 mg of an aminoglycoside.

51. The method of any one of items 30-44, wherein the pharmaceutical formulation comprises about 600 mg of an aminoglycoside.

52. A method of delivering a liposomal aminoglycoside aerosol, the method comprising

aerosolizing 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

The liposomal aminoglycoside aerosol is delivered to the patient's lungs via inhalation.

53. A method of delivering a liposomal aminoglycoside aerosol, the method comprising

aerosolizing about 8 to about 9 grams of an aqueous liposomal dispersion of an aminoglycoside for about 10 to about 15 minutes to obtain a liposomal aminoglycoside aerosol; and

The liposomal aminoglycoside aerosol is delivered to the patient's lungs via inhalation.

54. The method of claim 52 or 53, wherein the aqueous liposomal dispersion of the aminoglycoside is aerosolized 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.

55. The method of claim 52 or 53, wherein the aqueous liposomal dispersion of the aminoglycoside is aerosolized 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.

56. The method of any one of items 52-55, wherein the liposomal aminoglycoside aerosol comprises an aminoglycoside and liposomes, wherein the liposomes are composed of DPPC and cholesterol, wherein about 55% to about 75% of the aminoglycoside is liposomally complexed.

57. The method of any one of items 52-56, wherein the liposomal aminoglycoside aerosol has

an MMAD of about 3.2 μm to about 4.2 μm as measured by ACI, or an MMAD of about 4.4 μm to about 4.9 μm as measured by NGI;

a GSD of approximately 1.75 to approximately 1.80;

an FPF of greater than or equal to about 64% as measured by ACI, or an FPF of greater than or equal to about 51% as measured by NGI; or

FPD of about 35 to about 41.

58. The method of any one of items 52-57, wherein about 25% to about 35% of the liposomal aminoglycoside aerosol is deposited in the bronchial and alveolar regions of the patient's lungs.

59. The method of any one of items 52-58, wherein the aqueous liposomal dispersion of the aminoglycoside comprises the aminoglycoside and liposomes, the liposomes being composed of DPPC and cholesterol, wherein greater than about 95% of the aminoglycoside is encapsulated within the liposomes.

60. The method of any one of items 52-59, wherein the aqueous liposomal dispersion of the aminoglycoside comprises about 70 to about 75 mg/mL of the aminoglycoside; about 32 to about 35 mg/mL of DPPC; and about 16 mg/mL to about 17 mg/mL of cholesterol.

61. The method of any one of items 52-60, wherein the volume of the aqueous liposome dispersion is about 8 mL.

62. The method of any one of items 52-61, wherein the aminoglycoside is amikacin.

63. The method of any one of items 52-62, wherein the aminoglycoside is amikacin sulfate.

64. The method of any one of items 52-63, wherein about 500 mg to about 650 mg of the aminoglycoside is delivered to the patient's lungs via inhalation.

65. The method of any one of items 52-63, wherein about 550 mg to about 625 mg of the aminoglycoside is delivered to the patient's lungs via inhalation.

66. The method of any one of items 52-63, wherein about 550 mg to about 600 mg of the aminoglycoside is delivered to the patient's lungs via inhalation.

67. The method of any one of items 52-63, wherein about 560 mg of the aminoglycoside is delivered to the patient's lungs via inhalation.

68. The method of any one of items 52-63, wherein about 580 mg of the aminoglycoside is delivered to the patient's lungs via inhalation.

69. The method of any one of items 52-63, wherein about 590 mg of the aminoglycoside is delivered to the patient's lungs via inhalation.

70. The method of any one of items 52-63, wherein about 600 mg of the aminoglycoside is delivered to the patient's lungs by inhalation.

71. A liposomal aminoglycoside aerosol comprising an aminoglycoside and liposomes comprising DPPC and cholesterol, wherein about 65% to about 75% of the aminoglycoside is liposomally complexed and the liposomal aminoglycoside aerosol is generated at a rate greater than about 0.53 grams per minute.

72. The liposomal aminoglycoside aerosol of claim 71, which is generated at a rate of 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.

73. The liposomal aminoglycoside aerosol of claim 71 or 72, which is generated at a rate of about 0.60 to about 0.70 grams per minute.

74. The liposomal aminoglycoside aerosol of any one of items 71-73, which provides about 500 mg or about 560 mg of amikacin in a single dosing period.

75. The liposomal aminoglycoside aerosol of claim 74, wherein the administration period is once a day.

76. The liposomal aminoglycoside aerosol of 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.

77. The liposomal aminoglycoside aerosol of any one of items 71-76, wherein the MMAD of the aerosol is about 1.0 μm to about 4.2 μm as measured by ACI; about 2.0 μm to about 4.2 μm as measured by ACI; about 3.2 μm to about 4.2 μm as measured by ACI; about 1.0 μm to about 4.9 μm as measured by NGI; about 2.0 μm to about 4.9 μm as measured by NGI; about 4.4 μm to about 4.9 μm as measured by NGI.

78. The liposomal aminoglycoside aerosol of 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.

79. The liposomal aminoglycoside aerosol of any one of items 71-78, wherein the aerosol comprises greater than or equal to about 50% liposomally complexed amikacin.

80. The liposomal aminoglycoside aerosol of any one of items 71-79, wherein the aerosol comprises greater than or equal to about 60% liposomally complexed amikacin.

81. The liposomal aminoglycoside aerosol of any one of items 71-80, wherein the aerosol comprises about 55% to about 85% liposomally complexed amikacin.

82. The liposomal aminoglycoside aerosol of any one of items 71-81, wherein the aerosol comprises about 55% to about 75% liposomally complexed amikacin.

83. The liposomal aminoglycoside aerosol of any one of items 71 to 82, wherein the liposomes comprise unilamellar vesicles, multilamellar vesicles, or a mixture thereof.

84. The liposomal aminoglycoside aerosol of any one of items 71-83, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astamicin, kanamycin B, polmycin, tobramycin, capreomycin, dibekacin, datimicin, ethimicin, neomycin B, gentamicin, H107, hygromycin, hygromycin B, myosin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomicin, ribosomycin, sisomicin, rhodestreptomycin, sorbitol, spectinomycin, saccharopolysporin, streptomycin, tobramycin, methyl zisomycin, vetimicin, or a combination thereof.

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 liposomally complexed and the plurality of liposomes have a diameter of about 245 nm to about 290 nm as measured by light scattering.

86. The liposomal aminoglycoside aerosol of claim 85, which is generated at a rate greater than about 0.53 grams per minute.

87. The liposomal aminoglycoside aerosol of claim 85 or 86, which provides about 560 mg of amikacin over a dosing period.

88. The liposomal aminoglycoside aerosol of any one of items 85-87, wherein the administration period is once a day.

89. The liposomal aminoglycoside aerosol of any one of items 85-88, wherein the MMAD of the aerosol is about 1.0 μm to about 4.2 μm as measured by ACI; about 2.0 μm to about 4.2 μm as measured by ACI; about 3.2 μm to about 4.2 μm as measured by ACI; about 1.0 μm to about 4.9 μm as measured by NGI; about 2.0 μm to about 4.9 μm as measured by NGI; about 4.4 μm to about 4.9 μm as measured by NGI.

90. The liposomal aminoglycoside aerosol of any one of items 85-89, wherein the MMAD of the aerosol is about 3.2 μm to about 4.2 μm as measured by ACI, or about 4.4 μm to about 4.9 μm as measured by NGI.

91. The liposomal aminoglycoside aerosol of any one of items 85-90, 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.

92. The liposomal aminoglycoside aerosol of any one of items 85-91, wherein the aerosol comprises greater than about 55% liposomally complexed amikacin, greater than about 60% liposomally complexed amikacin, greater than about 65% liposomally complexed amikacin, or greater than about 70% liposomally complexed amikacin.

93. The liposomal aminoglycoside aerosol of any one of items 85-92, wherein the aerosol comprises about 55% to about 75% liposomally complexed amikacin.

94. The liposomal aminoglycoside aerosol of any one of items 85-93, wherein the liposomes comprise unilamellar vesicles, multilamellar vesicles, or a mixture thereof.

95. The liposomal aminoglycoside aerosol of any one of items 85-94, wherein the aminoglycoside is selected from amikacin, apramycin, arbekacin, astamicin, capreomycin, dibekacin, neomycin B, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribosomycin, sisomicin, spectinomycin, streptomycin, tobramycin, methyl zithromycin or a combination thereof.

96. The liposomal aminoglycoside aerosol of any one of items 71-95, which is generated at a rate of about 0.55 to about 0.70 grams per minute, or at a rate of about 0.60 to about 0.70 grams per minute, or at a rate of about 0.65 to about 0.70 grams per minute.

97. The liposomal aminoglycoside aerosol of any one of items 85-96, wherein the average liposome size measured by light scattering is about 265 nm.

98. The method of any one of items 30-51, wherein the patient has cystic fibrosis.

99. The method of any one of items 30-51 and 98, wherein the lung infection is a non-tuberculous mycobacterial infection.

100. The method of any one of items 30-51 and 98, wherein the lung infection is a Pseudomonas infection.

101. The method of any one of items 30-51 and 98, wherein the lung infection is a Burkholderia infection.

102. The method of claim 100, wherein the Pseudomonas infection is a Pseudomonas aeruginosa infection.

103. The method of claim 101, wherein the Burkholderia infection is caused by the following bacteria: Burkholderia pseudomallei (B. pseudomallei), Burkholderia cepaci (B. cepaci), Burkholderia cepacia complex (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. caribensisor, or B. caryophylli.

104. The method of claim 99, wherein the non-tuberculous mycobacterium infection is a Mycobacterium avium infection.

105. The method of claim 104, wherein the Mycobacterium avium infection is a Mycobacterium avium subsp. hominissuis infection.

106. The method of claim 99, wherein the non-tuberculous mycobacterium infection is a Mycobacterium abscessus infection.

107. The method of claim 99, wherein the non-tuberculous mycobacterium infection is a Mycobacterium avium complex (Mycobacterium avium and Mycobacterium intracellulare) infection.

108. The method of claim 99, wherein the non-tuberculous mycobacterial infection is selected from the group consisting of Mycobacterium avium (M. avium), Mycobacterium avium suis (MAH), Mycobacterium abscessus (M. abscessus), Mycobacterium chelonae (M. chelonae), Mycobacterium bolletii (M. bolletii), Mycobacterium kansasii (M. kansasii), Mycobacterium ulcerans (M. ulcerans), Mycobacterium avium, Mycobacterium avium complex (M. avium complex, MAC) (Mycobacterium avium and Mycobacterium intracellulare), Mycobacterium conspicuum, Mycobacterium kansasii, Mycobacterium peregrinum, Mycobacterium immunogenum, Mycobacterium xenopi, Mycobacterium marinum, Mycobacterium malmoense, Mycobacterium marinum, Mycobacterium mucogenicum, Mycobacterium nonchromogenicum, Mycobacterium scrofulaceum, Mycobacterium simiae, Mycobacterium smegmatis, Mycobacterium szulgai, Mycobacterium terrae, Mycobacterium terrae complex complex), M. haemophilum, M. genavense, M. asiaticum, M. shimoidei, M. gordonae, M. achromogenic, M. triplex, M. lentiflavum, M. celatum, M. fortuitum, M. fortuitum complex (M. fortuitum and M. chelonae), or a combination thereof.

109. The system of any one of items 1-29, wherein the concentration of the aminoglycoside is about 50 mg/mL or greater; or about 60 mg/mL or greater; or about 70 mg/mL or greater.

110. The system of any one of items 1-29, 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.

111. The system of any one of items 1-29, wherein the concentration of the aminoglycoside is about 60 mg/mL to about 80 mg/mL.

112. The system of any one of items 109-111, wherein the aminoglycoside is amikacin.

113. The system of claim 112, wherein the aminoglycoside is amikacin sulfate.

114. The method of any one of items 30-70 and 98-108, wherein the concentration of the aminoglycoside is about 50 mg/mL or greater; or about 60 mg/mL or greater; or about 70 mg/mL or greater.

115. The method of any one of items 30-70 and 98-108, 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.

116. The method of any one of items 30-70 and 98-108, wherein the concentration of the aminoglycoside is about 60 mg/mL to about 80 mg/mL.

117. The method of any one of items 30-70 and 98-108, wherein the concentration of the aminoglycoside is about 70 mg/mL to about 80 mg/mL.

118. The method of any one of items 115-117, wherein the aminoglycoside is amikacin.

119. The method of claim 118, wherein the aminoglycoside is amikacin sulfate.

120. The liposomal aminoglycoside aerosol of any one of items 71-97, wherein the concentration of the aminoglycoside is about 50 mg/mL or greater; or about 60 mg/mL or greater; or about 70 mg/mL or greater.

121. The liposomal aminoglycoside aerosol of any one of items 71-97, 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.

122. The liposomal aminoglycoside aerosol of any one of items 71-97, wherein the concentration of the aminoglycoside is about 60 mg/mL to about 80 mg/mL.

123. The liposomal aminoglycoside aerosol of any one of items 71-97, wherein the concentration of the aminoglycoside is about 70 mg/mL to about 80 mg/mL.

124. The liposomal aminoglycoside aerosol of any one of items 121 to 123, wherein the aminoglycoside is amikacin.

125. The liposomal aminoglycoside aerosol of claim 124, wherein the aminoglycoside is amikacin sulfate.

126. The system of any one of items 1-29, wherein the lung infection is a non-tuberculous mycobacterial lung infection.

127. The system of any one of items 1-29, wherein the lung infection is a Pseudomonas lung infection.

128. The system of any one of items 1-29, wherein the lung infection is a Burkholderia lung infection.

129. The system of claim 127, wherein the Pseudomonas infection is a Pseudomonas aeruginosa lung infection.

130. The system of claim 128, wherein the Burkholderia lung infection is a lung infection caused by Burkholderia pseudomallei, Burkholderia cepacia, Burkholderia cepacia complex, Burkholderia reluctance, Burkholderia fungi, Burkholderia gladiolus, Burkholderia multivora, Burkholderia vietnam, Burkholderia pseudomallei, Burkholderia afila, Burkholderia anthina, Burkholderia brasiliensis, Burkholderia caledonia, Burkholderia karyobensis, or Burkholderia dianthus.

131. The system of claim 126, wherein the non-tuberculous mycobacterial lung infection is a lung infection caused by Mycobacterium avium, Mycobacterium abscessus, Mycobacterium chelonae, Mycobacterium bovis, Mycobacterium kansasii, Mycobacterium ulcerans, Mycobacterium avium, Mycobacterium avium complex (MAC) (Mycobacterium avium and Mycobacterium intracellulare), Mycobacterium superiorum, Mycobacterium kansasii, Mycobacterium exogenum, Mycobacterium immunogenicum, Mycobacterium toad, Mycobacterium marinum, Mycobacterium malmoe, Mycobacterium marinum, Mycobacterium truncatum ... Mycobacterium, Mycobacterium mucogenes, Mycobacterium non-chromogenes, Mycobacterium scrofulae, Mycobacterium simiae, Mycobacterium smegmatis, Mycobacterium surga, Mycobacterium terrestris, Mycobacterium terrestris complex, Mycobacterium hemophilus, Mycobacterium geneva, Mycobacterium asiatica, Mycobacterium subspecies, Mycobacterium gordonii, Mycobacterium non-chromogenes, Mycobacterium tristis, Mycobacterium lentum, Mycobacterium occulta, Mycobacterium fortuitum, Mycobacterium fortuitum complex (Mycobacterium fortuitum and Mycobacterium chelonae), or a combination thereof.

132. The system of claim 131 , wherein the non-tuberculous mycobacterium lung infection is a Mycobacterium abscessus lung infection.

133. The system of claim 131 , wherein the non-tuberculous mycobacterium lung infection is a Mycobacterium avium lung infection.

134. The system of claim 133, wherein the non-tuberculous mycobacterium lung infection is a Mycobacterium avium subspecies hominis lung infection.

135. The method of any one of items 30-39, 41 and 44-70, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astamicin, kanamycin B, polmycin, tobramycin, capreomycin, dibekacin, datimicin, ethimicin, neomycin B, gentamicin, H107, hygromycin, hygromycin B, myosin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomicin, ribosomycin, sisomicin, rhodestreptomycin, sorbitol, spectinomycin, saccharopolysporin, streptomycin, tobramycin, methylthiazolin, vetimicin or a combination thereof.

136. The system of any one of items 1-3, 6-29, and 109-113, wherein the aminoglycoside is selected from AC4437, amikacin, apramycin, arbekacin, astamicin, kanamycin B, polmycin, tobramycin, capreomycin, dibekacin, datimicin, ethimicin, neomycin B, gentamicin, H107, hygromycin, hygromycin B, myosin, K-4619, isepamicin, KA-5685, kanamycin, neomycin, netilmicin, paromomycin, plazomicin, ribosomycin, sisomicin, rhodestreptomycin, sorbitol, spectinomycin, saccharopolysporin, streptomycin, tobramycin, methylthiazolin, vetimicin, or a combination thereof.

137. The system of any one of items 1-29, 109-113 and 136, wherein the aqueous dispersion is an aqueous suspension of a liposomally complexed aminoglycoside.

138. The system of any one of items 1-29, 109-113 and 136, wherein the aqueous dispersion is an aqueous solution of a liposomally complexed aminoglycoside.

139. The method of any one of items 30-70, 114-119 and 135, wherein the aqueous dispersion is an aqueous suspension of liposomally complexed aminoglycoside.

140. The method of any one of items 30-70, 114-119 and 135, wherein the aqueous dispersion is an aqueous solution of a liposomally complexed aminoglycoside.

141. The method of any one of items 30-51 and 98, wherein the lung infection is associated with bronchiectasis.

142. The method of claim 141, wherein the aqueous dispersion is an aqueous suspension of liposome-complexed aminoglycoside.

143. The method of claim 141, wherein the aqueous dispersion is an aqueous solution of a liposomally complexed aminoglycoside.

144. The system of any one of items 1-29, wherein the lung infection is associated with bronchiectasis.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 shows a diagram of a nebulizer (aerosol generator) in which the present invention may be implemented.

FIG. 2 is an enlarged illustration of the nebulizer diagram shown in FIG. 1 .

Figure 3 shows a cross-sectional view of a generally known aerosol generator as described in WO 2001/032246.

FIG4 is an image of a PARI nebulizer modified for use with the aminoglycoside formulations described herein, and a magnified view of the nebulizer membrane.

FIG5 is a cross-sectional computed tomography (CT) image showing a membrane having a relatively long nozzle portion.

FIG6 is a cross-sectional computed tomography (CT) image of a stainless steel membrane having a relatively short nozzle portion.

FIG. 7 is a schematic depiction of a cross-section of saliva/biofilm such as that found in cystic fibrosis patients.

8 is a graph showing the time period for aerosol generation (nebulization time) after complete release of liquid from a liquid reservoir as a function of the initial air cushion ( _VA ) within the liquid reservoir.

9 is a graph of negative pressure in the nebulizer as a function of the time from aerosol generation until complete release of the drug formulation from the liquid reservoir (nebulization time).

10 is a graph of aerosol generation efficiency as a function of negative pressure in the nebulizer.

11 is a graph of the time period for aerosol generation after complete release of liquid (nebulization time) as a function of the ratio (V _RN /V _L ) between the increased volume V _RN of the liquid reservoir and the initial volume of liquid in the liquid reservoir (V _L ).

FIG12 is a graph showing the MMAD of aerosolized formulations as a function of the nebulization rate of the corresponding formulations.

FIG13 is a graph of the FPF of aerosolized formulations as a function of the nebulization rate of the corresponding formulations.

Figure 14 is a schematic diagram of the system used for aerosol recovery after nebulization studies.

Detailed Description of the Invention

The invention described herein relates, in part, to administering aminoglycoside pharmaceutical formulations to the lungs of a subject, eg, for treating a lung disorder.

The term "treating" includes: (1) preventing or delaying the onset of clinical symptoms of a condition, disorder, or condition that develops in a subject who may be susceptible to or susceptible to the condition, disorder, or condition, but who does not yet experience or express clinical or subclinical symptoms of the condition, disorder, or condition; (2) inhibiting the condition, disorder, or condition (i.e., arresting, reducing, or delaying the development of the disease or its recurrence, arresting, reducing, or delaying the development of at least one clinical or subclinical symptom thereof, in the case of maintenance treatment); and/or (3) alleviating the condition (i.e., causing regression of the condition, disorder, or condition, or at least one clinical or subclinical symptom thereof). The benefit to the subject being treated is statistically significant or at least perceptible to the subject or physician.

In one embodiment, the systems and formulations provided herein can be used to 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), complex), Burkholderia dolosa, B. fungorum, B. gladioli, B. multivorans, B. vietnamiensis, Burkholderia pseudomallei, B. ambifaria, B. andropogonis, B. anthina, B. brasilensis, B. caledonia ledonica), Burkholderia caribensis, Burkholderia caryophylli), Staphylococcus (e.g., S. aureus, S. auricularis, S. carnosus, S. epidermidis, S. lugdunensis), Methicillin-resistant Staphylococcus aureus Staphylococcus aureus (MRSA), Streptococcus (e.g., Streptococcus pneumoniae), Escherichia coli, Klebsiella, Enterobacter, Serratia, Haemophilus, Yersinia pestis, Mycobacterium (e.g., nontuberculous mycobacteria).

In one embodiment, a system provided herein is used to treat a patient's nontuberculous mycobacterial lung infection. In a further embodiment, the nontuberculous mycobacterial lung infection is a refractory nontuberculous mycobacterial lung infection.

In one embodiment, the system provided herein is used to treat a patient having a lung infection caused by Pseudomonas. In a further embodiment, the lung infection is caused by a Pseudomonas species selected from the species provided in Table B below.

In one embodiment, the nontuberculous mycobacterial lung infection is selected from the group consisting of Mycobacterium avium, Mycobacterium avium hominis subsp. suis (MAH), Mycobacterium abscessus, Mycobacterium chelonae, Mycobacterium bovis, Mycobacterium kansasii, Mycobacterium ulcerans, Mycobacterium avium, Mycobacterium avium complex (MAC) (Mycobacterium avium and Mycobacterium intracellulare), Mycobacterium spp., Mycobacterium kansasii, Mycobacterium exogenum, Mycobacterium immunogenicum, Mycobacterium toad, Mycobacterium marinum, Mycobacterium malmoe, Mycobacterium marinum, Mycobacterium truncatum ... In some embodiments, the nontuberculous mycobacterium is infected with Mycobacterium spp., Mycobacterium mucogenicum, Mycobacterium achromogenicum, Mycobacterium scrofulaceum, Mycobacterium simiae, Mycobacterium smegmatis, Mycobacterium surga, Mycobacterium terrestris, Mycobacterium terrestris complex, Mycobacterium hemophilus, Mycobacterium geneva, Mycobacterium asiatica, Mycobacterium sulphureus, Mycobacterium gordonii, Mycobacterium achromogenicum, Mycobacterium trifum, Mycobacterium lentum, Mycobacterium crypticum, Mycobacterium fortuitum, Mycobacterium fortuitum complex (Mycobacterium fortuitum and Mycobacterium chelonae), or a combination thereof. In a further embodiment, the nontuberculous mycobacterium lung infection is Mycobacterium abscessus or Mycobacterium avium. In a further embodiment, the Mycobacterium avium infection is Mycobacterium avium subspecies Hominissuis. In one embodiment, the nontuberculous mycobacterium lung infection is a refractory nontuberculous mycobacterium lung infection.

In another embodiment, a system provided herein is used to treat a bacterial infection in a cystic fibrosis patient. In a further embodiment, the bacterial infection is a lung infection due to Pseudomonas aeruginosa. In yet another embodiment, a system provided herein is used to treat a lung infection associated with bronchiectasis in a patient.

As used herein, "prevent" or "prevent" may refer to preventing an infection or disease entirely, or preventing the development of symptoms of the infection or disease; delaying the onset of an infection or disease or its symptoms; or reducing the severity of an infection or disease or its symptoms that subsequently develops.

The term "antibacterial" is art-recognized and refers to the ability of the compounds of the present invention to prevent, inhibit or destroy the growth of bacterial microorganisms. Examples of bacteria are provided above.

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

An "effective amount" refers to the amount of an aminoglycoside (e.g., amikacin) used in the present invention that is sufficient to produce the desired therapeutic response. An effective amount of the formulations provided herein comprises both free and liposome-complexed aminoglycosides. For example, in one embodiment, the liposome-complexed aminoglycoside comprises an aminoglycoside encapsulated in or complexed with liposomes, or a combination thereof.

In one embodiment, the aminoglycoside is selected from amikacin, apramycin, arbekacin, astamicin, capreomycin, dibekacin, neomycin B, gentamicin, hygromycin B, isepamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribosomycin, sisomicin, spectinomycin, streptomycin, tobramycin or methyl zithromycin. In another embodiment, the aminoglycoside is selected from the aminoglycosides listed in Table C below.

In one embodiment, the aminoglycoside is an aminoglycoside free base or a salt, solvate, or other non-covalent derivative thereof. In a further embodiment, the aminoglycoside is amikacin. Suitable aminoglycosides for use in the pharmaceutical formulations of the present invention include pharmaceutically acceptable addition salts and complexes of the drug. Where the compound may have one or more chiral centers, unless otherwise specified, the present invention encompasses each specific racemic compound as well as each specific non-racemic compound. Where the 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 the active agent exists in tautomeric forms (e.g., keto-enol tautomers), each tautomeric form is contemplated to be included in the present invention. In one embodiment, amikacin is present in the pharmaceutical formulation as amikacin base or an amikacin salt, such as amikacin sulfate or amikacin bisulfate. In one embodiment, a combination of one or more of the above-described aminoglycosides is used in the formulations, systems, and methods described herein. In a further embodiment, the combination comprises amikacin.

A therapeutic response can be any response that a user (e.g., a clinician) recognizes as an effective response to treatment. A therapeutic response will typically be a reduction, inhibition, delay, or prevention of the growth or reproduction of one or more bacteria, or the killing of one or more bacteria, as described above. A therapeutic response can also be reflected in improvements in lung function, such as forced expiratory volume in one second (FEV ₁ ). Based on the assessment of the therapeutic response, it is further within the skill of those skilled in the art to determine the appropriate duration of treatment, appropriate dosage, and any potential combination therapies.

"Liposomal dispersion" refers to a solution or suspension comprising a plurality of liposomes.

As used herein, an "aerosol" is a gaseous suspension of liquid particles. The aerosol provided herein comprises particles of a liposome dispersion.

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. Pulmonary, ultrasonic, and electronic nebulizers, such as passive electronic mesh nebulizers, active electronic mesh nebulizers, and vibrating mesh nebulizers, are suitable for use with the present invention if the particular nebulizer emits an aerosol with the desired properties at the desired output rate.

The process of large volume (bulk) liquid being pneumatically converted into small droplets is called atomization.The operation of pneumatic nebulizer needs pressurized gas supply as the driving force of liquid atomization.Ultrasonic nebulizer uses the electric current introduced by the piezoelectric element in the liquid reservoir to convert liquid into absorbable droplets.Various types of nebulizers are described in Respiratory Care, Vol. 45, No. 6, pp. 609-622 (2000), the disclosure of which is incorporated into the present application in its entirety by reference.Term " nebulizer " and " aerosol generator " are used interchangeably throughout the specification." inhalation device ", " inhalation system " and " nebulizer " also can be used interchangeably with term " nebulizer " and " aerosol generator " in the present application.

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. The FPF is typically expressed as a percentage.

"Mass median diameter" or "MMD" is determined by laser diffraction or impactor measurement and is the mass average particle size.

"Total median aerodynamic diameter" or "MMAD" is standardized for aerodynamic separation of aqueous aerosol droplets and is determined by impactor measurements, e.g., an Anderson Cascade Impactor (ACI) or a New Generation Impactor (NGI). In one embodiment, the gas flow rate is 28 liters per minute (via an Anderson Cascade Impactor (ACI)) and 15 liters per minute (via a New Generation Impactor (NGI)). "Geometric standard deviation" or "GSD" is a measure of the distribution of aerodynamic particle size distribution.

In one embodiment, the present invention provides a system for treating lung infection or providing prevention of lung infection. Treatment is accomplished by delivering an aminoglycoside formulation via nebulized inhalation. In one embodiment, the pharmaceutical formulation comprises an aminoglycoside agent, e.g., an aminoglycoside.

As provided herein, the pharmaceutical formulations are liposomal dispersions. In particular, the pharmaceutical formulations are dispersions comprising "liposomally complexed aminoglycosides" or "aminoglycosides encapsulated in liposomes." "Liposomally complexed aminoglycosides" include embodiments in which the aminoglycoside (or combination of aminoglycosides) is encapsulated in liposomes, and include any form of aminoglycoside composition in which at least about 1% by weight of the aminoglycoside is associated with the liposomes, as part of a liposome complex, or as a liposome in which the aminoglycoside is in the aqueous phase or the hydrophobic bilayer phase or in the interfacial headgroup region of the liposome bilayer.

In one embodiment, the lipid component of the liposome comprises neutral lipids, positively charged lipids, negatively charged lipids, or a combination thereof. In another embodiment, the lipid component comprises neutral lipids. In a further embodiment, the lipid component consists essentially of neutral lipids. In yet further embodiments, the lipid component consists of neutral lipids, e.g., sterols and phospholipids.

Liposomally complexed aminoglycoside embodiments as provided above include embodiments in which the aminoglycoside is encapsulated in liposomes. Additionally, liposomally complexed aminoglycosides describe 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 liposome complex or as a liposome in which the aminoglycoside can be in the aqueous phase or the hydrophobic bilayer phase or in the interfacial headgroup region of the liposome bilayer. In one embodiment, prior to spraying, 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. In one embodiment, association is measured by filter separation, in which the lipids and lipid-bound drug are retained (i.e., in the retentate) and the free drug is in the filtrate.

The formulations, systems and methods provided herein comprise lipid-encapsulated or lipid-bound aminoglycosides. The lipids used in the pharmaceutical formulations of the present invention can be synthetic, semisynthetic or naturally occurring lipids, including phospholipids, tocopherols, sterols, fatty acids, negatively charged lipids and cationic lipids.

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

In one embodiment, the pharmaceutical formulation includes dipalmitoylphosphatidylcholine (DPPC), which is a major component of naturally occurring lung surfactant. In one embodiment, the lipid component of the pharmaceutical formulation comprises DPPC and cholesterol, or consists essentially of DPPC and cholesterol, or consists of DPPC and cholesterol. In a further embodiment, the molar ratio of DPPC and cholesterol ranges 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 further embodiments, the molar ratio of DPPC and cholesterol is about 2:1 or about 1:1. In one embodiment, DPPC and cholesterol are provided in an aminoglycoside formulation, e.g., an aminoglycoside formulation.

Other examples of lipids for use in the present invention include, but are not limited to, dimyristoylphosphatidylcholine (DMPC), dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylcholine (DSPC), distearoylphosphatidylglycerol (DSPG), dioleoylphosphatidylethanolamine (DOPE), mixed phospholipids such as palmitoylstearoylphosphatidylcholine (PSPC), and monoacylated phospholipids, e.g., monooleoylphosphatidylethanolamine (MOPE).

In one embodiment, at least one lipid component comprises sterol. In a further embodiment, at least one lipid component comprises sterol and phospholipid, or is basically composed of sterol and phospholipid, or is composed of sterol and phospholipid. Sterol for the present invention includes but is not limited to cholesterol, cholesterol ester including cholesterol hemisuccinate, cholesterol salt including cholesterol bisulfate and cholesterol sulfate, ergosterol, ergosterol ester including ergosterol hemisuccinate, ergosterol salt including ergosterol bisulfate and ergosterol sulfate, lanosterol, lanosterol ester including lanosterol hemisuccinate, lanosterol salt including lanosterol bisulfate and lanosterol sulfate and tocopherol. Tocopherol includes tocopherol, tocopherol ester including tocopherol hemisuccinate, tocopherol salt including tocopherol bisulfate and tocopherol sulfate. The term "sterol compound" includes sterol, tocopherol etc.

In one embodiment, at least one cationic lipid (positively charged lipid) is provided in the system described in the present application. Cationic lipid used can include the ammonium salt of fatty acid, phospholipid and glyceride. Fatty acid includes fatty acid with a carbon chain length of 12-26 carbon atoms, which is saturated or unsaturated. Some specific examples include: myristylamine, palmitylamine, laurylamine and stearamide, dilauroylethylphosphocholine (DLEP), dimyristoylethylphosphocholine (DMEP), dipalmitoylethylphosphocholine (DPEP) and distearoylethylphosphocholine (DSEP), N-(2,3-bis-(9-(Z)-octadecene base oxygen base)-propyl-1-base-N, N, N-trimethylammonium chloride (DOTMA) and 1,2-bis-(oleoyloxy)-3-(trimethylammonium) propane (DOTAP).

In one embodiment, at least one anionic lipid (negatively charged lipid) is provided in the system described herein. Operable negatively charged lipids include phosphatidylglycerols (PGs), phosphatidic acids (PAs), phosphatidylinositols (PIs), and phosphatidylserines (PSs). Examples include DMPG, DPPG, DSPG, DMPA, DPPA, DSPA, DMPI, DPPI, DSPI, DMPS, DPPS, and DSPS.

Without wishing to be bound by theory, phosphatidylcholine (e.g., DPPC) helps cells in the lungs (e.g., alveolar macrophages) to take up aminoglycosides and helps maintain the aminoglycosides in the lungs. It is believed that negatively charged lipids such as PGs, PAs, PSs, and PIs, in addition to reducing particle aggregation, also play a role in the sustained activity characteristics of inhaled formulations and the transpulmonary transport (transcytosis) of formulations for systemic uptake. Without wishing to be bound by theory, it is believed that sterol compounds affect the release characteristics of formulations.

Liposomes are completely closed lipid bilayer membranes containing an encapsulated aqueous volume. 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 having hydrophobic "tail" regions and hydrophilic "head" regions. The structure of the membrane bilayer is such that the hydrophobic (non-polar) "tails" of the lipid monolayers are oriented toward the center of the bilayer, while the hydrophilic "heads" are oriented toward the aqueous phase.

Liposome can be prepared by several methods (referring to, for example, Cullis etc. (1987)).In one embodiment, one or more methods described in U.S. Patent Application Publication 2008/0089927 are used to prepare the lipid preparation (liposomal dispersion) of aminoglycoside encapsulation in the present application.The disclosure of U.S. Patent Application Publication 2008/0089927 is incorporated by reference in its entirety for all purposes.For example, in one embodiment, at least one lipid and aminoglycoside are mixed with coacervate (that is, independent liquid phase) to form liposome preparation.Coacervate can be formed before mixing with lipid, during mixing with lipid or after mixing with lipid.In addition, coacervate can be the coacervate of activating agent.

In one embodiment, the liposome dispersion is formed by dissolving one or more lipids in an organic solvent to form a lipid solution, and mixing an aqueous solution of aminoglycoside with the lipid solution to form aminoglycoside aggregates. In a further embodiment, the organic solvent is ethanol. In yet a further embodiment, the one or more lipids comprise a phospholipid and a sterol.

In one embodiment, liposomes are prepared by sonication, extrusion, homogenization, swelling, electroforming, inverse emulsion, or reverse evaporation. Bangham's procedure (J. Mol. Biol. (1965)) prepared 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) disclosed methods for preparing multilamellar vesicles having substantially equal interlamellar solute distribution in their respective aqueous compartments. Paphadjopoulos et al., U.S. Pat. No. 4,235,871, disclosed methods for preparing paucilamellar liposomes by reverse phase evaporation. Each method is suitable for use in the present invention.

Unilamellar vesicles can be prepared from MLVs by a number of techniques, for example, the extrusion techniques of U.S. Pat. No. 5,008,050 and U.S. Pat. No. 5,059,421. Thus, sonication and homogenization can be used to prepare smaller unilamellar vesicles from larger liposomes (see, for example, Paphadjopoulos et al. (1968); Deamer and Uster (1983); and Chapman et al. (1968)).

The liposome preparation method of Bangham et al. (J. Mol. Biol. 13, 1965, pp. 238-252) involves suspending phospholipids in an organic solvent and then evaporating to dryness, leaving a thin film of phospholipids on the reaction vessel. An appropriate amount of aqueous phase is then added, "swelling" for 60 minutes, and the resulting liposomes, consisting of multilamellar vesicles (MLVs), are mechanically dispersed. This preparation method provided the basis for the development of small, ultrasonically produced unilamellar vesicles and large unilamellar vesicles described by Papahadjopoulos et al. (Biochim. Biophys. Acta. 135, 1967, pp. 624-638).

Techniques for preparing large unilamellar vesicles (LUVs), such as reverse phase evaporation, immersion procedures, and detergent dilution methods, can be used to prepare liposomes for use in the pharmaceutical formulations provided herein. A review of these and other methods for preparing liposomes can be found in the text Liposomes, Marc Ostro, ed., Marcel Dekker, Inc., New York, 1983, Chapter 1, which is incorporated herein by reference. See also Szoka, Jr. et al., (Ann. Rev. Biophys. Bioeng. 9, 1980, p. 467), which is also incorporated by reference in its entirety for all purposes.

Other techniques for preparing liposomes include those that form reverse-evaporation vesicles (REVs), U.S. Pat. No. 4,235,871. Another class of liposomes that can be used is characterized by having a substantially equal lamellar solute distribution. This class of liposomes is designated as stable multilamellar vesicles (SPLVs), as defined in U.S. Pat. No. 4,522,803, and includes unilamellar liposomes as described in U.S. Pat. No. 4,588,578 and frozen and thawed multilamellar vesicles (FATMLVs) as described above.

Various sterols and their water-soluble derivatives, such as cholesterol hemisuccinate, have been used to form liposomes; see, for example, U.S. Patent No. 4,721,612. Mayhew et al., PCT Publication WO 85/00968, describe methods for reducing drug toxicity by encapsulating the drug in liposomes containing α-tocopherol and certain derivatives thereof. In addition, various tocopherols and their water-soluble derivatives have been used to form liposomes; see PCT Publication 87/02219.

In one embodiment, it is about 0.01 micron to about 3.0 microns that the pharmaceutical preparation comprises mean diameter (measured by light scattering method) before spraying, for example, the scope is the liposome of about 0.2 to about 1.0 microns.In one embodiment, the mean diameter of the liposome in the preparation is about 200nm to about 300nm, about 210nm to about 290nm, about 220nm to about 280nm, about 230nm to about 280nm, about 240nm to about 280nm, about 250nm to about 280nm or about 260nm to about 280nm.The sustained activity curve of liposome product can be regulated by the character of lipid membrane and by other excipients included in the composition.

In order to minimize the dosage volume and reduce the administration time of the patient, in one embodiment, it is important that the liposome encapsulation of the aminoglycoside (e.g., aminoglycoside amikacin) is efficient and the L/D ratio is as low as possible and/or practically possible while keeping the liposomes small enough to penetrate the patient's mucus and biofilm, for example, in the case of Pseudomonas biofilm. In one embodiment, the L/D ratio of 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 a further embodiment, the average diameter of the liposomes (as measured by light scattering) is from about 260 to about 280 nm.

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

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

In one embodiment, the system provided herein comprises an aminoglycoside formulation, for example, 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.

In one embodiment, the system provided herein comprises about 8 mL of a liposomal amikacin formulation. In one embodiment, the liposomal amikacin formulation has a density of about 1.05 g/mL; and in one embodiment, approximately 8.4 g 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.

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 the formulation provided in Table 2 below.

It should be noted that increasing aminoglycoside concentration alone may not result in a decrease in dosing time. For example, in one embodiment, the lipid to drug ratio is fixed, and as the amikacin concentration increases (since the ratio of the two is fixed, the lipid concentration increases, e.g., at ~0.7:1), the solution viscosity also increases, which slows the nebulization time.

In one embodiment, prior to nebulization of an aminoglycoside formulation, about 70% to about 100% of the aminoglycoside present in the formulation is liposomally complexed. In a further embodiment, the aminoglycoside is an aminoglycoside. In a still further 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 liposomally complexed. In a further embodiment, the aminoglycoside is amikacin or tobramycin. In a still further embodiment, the aminoglycoside is amikacin. In another embodiment, prior to nebulization, about 98% of the aminoglycoside present in the formulation is liposomally complexed. In a further embodiment, the aminoglycoside is amikacin or tobramycin. In a still further embodiment, the aminoglycoside is amikacin.

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

As provided herein, the present invention provides methods and systems for treating lung infections by inhalation of a liposomal aminoglycoside formulation. In one embodiment, the formulation is administered via a nebulizer that provides an aerosol spray of the formulation for delivery to the lungs of a subject.

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

After nebulization, the liposomes in the drug formulation leak the drug. In one embodiment, the amount of aminoglycoside complexed by the liposomes 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 the "percentage of aminoglycoside bound after nebulization." As provided herein, in one embodiment, the liposomes comprise an aminoglycoside, e.g., amikacin. In one embodiment, the percentage of aminoglycoside bound after nebulization is about 60% to about 70%. In a further embodiment, the aminoglycoside is amikacin. In another embodiment, the percentage of aminoglycoside bound after nebulization is about 67%, or about 65% to about 70%. In a further embodiment, the aminoglycoside is amikacin.

In one embodiment, the percentage of aminoglycoside bound after nebulization is measured by recovering the aerosol from the air condensed in a cold trap and subsequently determining the liquid for free and encapsulated aminoglycoside (bound aminoglycoside).

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 an ACI at a gas flow rate of about 28 L/min or by a new generation impactor, NGI, at a gas flow rate of about 15 L/min.

In one embodiment, the MMAD of the aerosol of the pharmaceutical formulation is measured by ACI and 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. In one embodiment, the MMAD of the aerosol of the pharmaceutical formulation is measured by NGI and 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.

In another embodiment, the nebulizer described herein generates an aerosol of an aminoglycoside pharmaceutical formulation at a rate of greater than about 0.53 g per minute, greater than about 0.55 g per minute, or greater than about 0.60 g per minute, or about 0.60 g per minute to about 0.70 g per minute. In further embodiments, the aerosol has an FPF of 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.

In one embodiment, the system provided herein comprises a nebulizer selected from the group consisting of an electronic mesh nebulizer, a pulmonary (jet) nebulizer, an ultrasonic nebulizer, a breath-enhanced nebulizer, and a breath-actuated nebulizer. In one embodiment, the nebulizer is portable.

The operating principle of a pulmonary nebulizer is generally known to those of ordinary skill in the art and is described in, for example, Respiratory Care, Vol. 45, No. 6, pp. 609-622 (2000). In short, a pressurized gas supply is used as the driving force for liquid atomization in a pneumatic nebulizer. Compressed gas is delivered, which causes a negative pressure zone. The solution to be aerosolized is then delivered into the air flow and sheared into a liquid film. The film is unstable and breaks into droplets due to surface tension. Smaller particles, i.e., particles with the above-mentioned MMAD and FPF properties, can then be formed by placing a baffle in the aerosol stream. In one embodiment of a pulmonary nebulizer, the gas and solution are mixed before leaving the outlet orifice (nozzle) and interacting with the baffle. In another embodiment, mixing does not occur until the liquid and gas leave the outlet orifice (nozzle). In one embodiment, the gas is air, O ₂ and/or CO ₂ .

In one embodiment, the droplet size and output rate can be customized in a pulmonary nebulizer. However, it should be considered whether the properties of the formulation of the spray and the formulation (e.g., % bound aminoglycoside) have changed due to the modification of the nebulizer. For example, in one embodiment, the gas velocity and/or pharmaceutical formulation speed are modified to achieve the output rate and droplet size of the present invention. Additionally or alternatively, the flow rate of the gas and/or solution can be customized to achieve the droplet size and output rate of the present invention. For example, in one embodiment, an increase in the gas velocity reduces the droplet size. In one embodiment, the ratio of the pharmaceutical formulation flow to the gas flow can be customized to achieve the droplet size and output rate of the present invention. In one embodiment, an increase in the ratio of liquid to gas flow increases the particle size.

In one embodiment, the pulmonary nebulizer output rate is increased by increasing the fill volume in the liquid reservoir. Without wishing to be bound by theory, the increase in output rate may be due to a decrease in dead volume in the nebulizer. In one embodiment, the nebulization time is reduced by increasing the power flow to the nebulizer. See, for example, Clay et al. (1983). Lancet 2, pp. 592-594 and Hess et al. (1996). Chest 110, pp. 498-505.

In one embodiment, a storage bag is used to capture the aerosol during the nebulization process and subsequently provide the aerosol to the subject via inhalation. In another embodiment, the nebulizer provided herein includes an open hole design equipped with a valve. In this embodiment, the nebulizer output increases as the patient inhales through the nebulizer. During the exhalation phase, a one-way valve diverts the patient's flow away from the nebulizing chamber.

In one embodiment, the nebulizer provided herein is a continuous nebulizer. In other words, when a dose is given, there is no need to refill the nebulizer with the pharmaceutical formulation. Instead, the nebulizer has a capacity of at least 8 mL or at least 10 mL.

In one embodiment, a vibrating mesh nebulizer is used to deliver an aminoglycoside formulation of the 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, about 110 kHz to about 150 kHz. In one embodiment, the nebulizer membrane vibrates at a frequency of about 117 kHz upon application of an electric current.

In one embodiment, the nebulizer provided herein does not use an air compressor and therefore does not generate an air stream. 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 rear of the mixing chamber and delivers the aerosol to the patient through the mouthpiece. When the patient exhales, the patient's respiratory flow passes through the one-way exhalation valve on the device mouthpiece. In one embodiment, the nebulizer continues to generate aerosol that enters the mixing chamber and is then inhaled by the subject's next breath - and this cycle continues until the nebulizer drug reservoir is empty.

Although not limited thereto, in one embodiment, the present invention is implemented using an aerosol generator (nebulizer) as depicted in Figures 1, 2, 3, and 4. Furthermore, in one embodiment, the system of the present invention includes a nebulizer as described in European Patent Application Nos. 11169080.6 and/or 10192385.2. These applications are incorporated herein by reference in their entirety.

Figure 1 shows a therapeutic aerosol device 1 having a nebulization chamber 2, a mouthpiece 3 and a membrane aerosol generator 4 having an oscillating membrane 5. The oscillating membrane may be oscillated, for example, by a ring-shaped piezoelectric element (not shown), an example of which is described in WO 1997/29851.

During use, the drug formulation is located on one side of the oscillating membrane 5, see Figures 1, 2 and 4, and then the liquid is transported through the opening in the oscillating membrane 5 and released on the other side of the oscillating membrane 5, see the bottom of Figures 1 and 2, and enters the nebulization chamber 2 in the form of an aerosol. The patient can inhale the aerosol present in the nebulization chamber 2 at the mouthpiece 3.

Oscillating membrane 5 comprises a plurality of through holes.When aminoglycoside pharmaceutical preparation passes through membrane, the droplet of aminoglycoside preparation is generated.In one embodiment, membrane is vibrable, i.e. so-called active electronic mesh sprayer, for example the sprayer of PARI Pharma, the HL100 sprayer of Health and Life or the Aeroneb of Aerogen (Novartis) in a further embodiment, membrane vibrates with the ultrasonic frequency of about 100kHz to about 150kHz, about 110kHz to about 140kHz or about 110kHz to about 120kHz.In a further embodiment, membrane vibrates with the frequency of about 117kHz after applying electric current.In a further embodiment, membrane is fixed and another part of liquid reservoir or liquid supply is vibrable, i.e. so-called passive electronic mesh sprayer, for example the MicroAir electronic sprayer model U22 of Omron or the I-Neb I-neb AAD inhalation system of Philips Respironics.

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

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

In one embodiment, the length of the nozzle section affects the geometric standard deviation (GSD) of the droplet size distribution of the aminoglycoside pharmaceutical preparation. Low GSD is characterized by a narrow droplet size distribution (droplets of uniform size), which is advantageous for targeting aerosols 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 section, the lower the GSD. In one embodiment, the average droplet size is less than 5 μm, and the scope of GSD is 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.

In one embodiment, as provided above, the system provided herein comprises a nebulizer that generates an aerosol of an aminoglycoside pharmaceutical formulation at a rate of greater than about 0.53 g per minute or greater than about 0.55 g per minute. In a further embodiment, the nebulizer comprises a vibrating membrane having a first side in contact with the liquid and an opposing second side, wherein droplets emerge from the second side.

Film (for example, stainless steel film) can be vibrated by piezoelectric actuator or any other suitable means.Film has a plurality of through holes that run through film in the direction of extension from the first side to the second side.Through hole can be formed by laser source, electroforming or any other suitable method as mentioned above.When film vibrates, aminoglycoside pharmaceutical preparation passes through hole from the first side to the second side to generate aerosol on the second side.In one embodiment, each through hole comprises inlet opening and outlet opening.In a further embodiment, each through hole comprises the nozzle portion extending towards the inlet opening through a part of through hole from outlet opening.Nozzle portion is limited by the continuous part of the through hole on the direction of extension that comprises the minimum diameter of through hole, and forms a boundary with the larger diameter of through hole.In one embodiment, the larger diameter of through hole is defined as 3 times, about 3 times, 2 times, about 2 times, 1.5 times or about 1.5 times the diameter closest to the minimum diameter.

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

In one embodiment, the sprayer provided herein comprises a through-hole, wherein the ratio of the total length of at least one through-hole in the extension direction to the length of the corresponding nozzle portion of the through-hole in the extension direction is at least 4, or at least about 4, or at least 4.5, or at least about 4.5, or at least 5, or at least about 5, or greater than about 5. In another embodiment, the sprayer provided herein comprises a through-hole, wherein the ratio of the total length of the majority of the through-holes in the extension direction to the length of the corresponding nozzle portion of the through-hole in the extension direction is at least 4, or at least about 4, or at least 4.5, or at least about 4.5, or at least 5, or at least about 5, or greater than about 5.

In one embodiment, the extended ratio described above provides an increased total output rate compared to previously known nebulizers while also providing adequate GSD. In one embodiment, the ratio structure enables a shorter application time, resulting in greater patient comfort and effectiveness of the aminoglycoside compound. This is particularly advantageous if, due to its nature, the aminoglycoside compound in the formulation is prepared at a low concentration, and therefore a larger volume of the aminoglycoside pharmaceutical formulation must be administered within an acceptable time (e.g., one dosing period).

According to one embodiment, the nozzle portion terminates flush with the second side. Thus, in one embodiment, the length of the nozzle portion is defined as the portion extending from the second side toward the first side to a diameter that is approximately three times, approximately two times, approximately 2.5 times, or approximately 1.5 times the minimum diameter. In this embodiment, the minimum diameter is the diameter of the outlet opening.

In one embodiment, the smallest diameter (i.e., one side of the nozzle portion) is located at the end of the nozzle portion in the direction of extension adjacent the second side. In one embodiment, the larger diameter of the through-holes located on the other side of the nozzle portion is located upstream of the smallest diameter in the direction of liquid passing through the plurality of through-holes during operation.

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.

In one embodiment, the total length of at least one through-hole in the direction of extension 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 a majority of the plurality of through-holes in the direction of extension 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 a majority of the plurality of through-holes is at least about 90 μm.

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

According to one embodiment, the through hole is a laser drilled through hole formed in at least two stages, one stage to form the nozzle portion and another stage to form the remainder of the through hole.

In another embodiment, the manufacturing method used results in a substantially cylindrical or conical nozzle portion having 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.

Alternatively or additionally, the through-hole is formed by electroforming. In one embodiment, the through-hole has a first funnel-shaped portion on a first side, a second funnel-shaped portion on a second side, and a nozzle portion intermediate the first and second funnel-shaped portions, defined between the outlet opening and the larger diameter. In this case, the total length of the through-hole can also be defined solely by the distance from the first side to the outlet opening (minimum diameter).

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

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

In one embodiment, the membrane comprises from about 200 to about 8,000 through holes, from about 1,000 to about 6,000 through holes, from about 2,000 to about 5,000 through holes, or from about 2,000 to about 4,000 through holes. In one embodiment, the number of through holes increases the TOR, and the TOR increases regardless of whether the nozzle parameters are implemented as described above. In one embodiment, the sprayer provided herein comprises about 3,000 through holes. In a further embodiment, the through holes are located in a hexagonal array, for example, at approximately 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.

Figure 3 shows an aerosol generator (nebulizer) as disclosed in WO 2001/032246, incorporated herein by reference in its entirety.The aerosol generator comprises a liquid container 21 to contain a drug formulation, emitted as an aerosol into a mixing chamber 3 and inhaled through a mouthpiece 4 via an opening 41 .

The aerosol generator includes a vibrating membrane 22 that is vibrated by a piezoelectric actuator 23. The vibrating membrane 22 has a first side 24 facing the liquid container 21 and a second, opposite side 25 facing the mixing chamber 3. In use, the first side 24 of the vibrating membrane 22 comes into contact with the liquid contained in the liquid container 21. A plurality of through-holes 26 are provided in the membrane 22, extending from the first side 24 to the second side 25. In use, when the membrane 22 vibrates, liquid passes from the liquid container 21 through the through-holes 26 from the first side 24 to the second side 25 to generate an aerosol at the second side 25 and emit it into the mixing chamber 3. The aerosol can then be inhaled by the patient through the mouthpiece 4 and its inhalation port 41 from the mixing chamber 3.

FIG5 shows a cross-sectional computed tomography scan showing three through-holes 26 of this vibrating membrane 22. The through-holes 26 of this embodiment are formed by laser drilling using different process parameters in three stages. In the first stage, portion 30 is formed. In the second stage, portion 31 is formed, and in the third stage, nozzle portion 32 is formed. In this embodiment, the length of nozzle portion 32 is about 26 μm, while 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 portion 30, portion 31 and nozzle portion 32, which in this specific example is about 101.5 μm. Therefore, the ratio of the total length of each through-hole 26 in the extension direction E to the length of a corresponding nozzle portion 32 in the extension direction E is about 3.9.

In the embodiment of FIG6 , first portion 30 has a length of approximately 27 μm, portion 31 has a length of approximately 55 μm, and the nozzle portion has a length of approximately 19 μm. As a result, the total length of through-hole 26 is approximately 101 μm. Therefore, 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.

The vibrating membranes in Figures 5 and 6 were each made with 6,000 through-holes 26. The following table (Table 3) shows the mass median diameter (MMD) of the particles emitted at the second side of the membrane (as measured by laser diffraction), the time required to completely emit a certain amount of liquid (spray time), and the TOR. The tests were performed using a liposomal formulation of amikacin.

Table 3 shows that Membrane 2, having a shorter nozzle section, provided an increased TOR and a reduced spray time of 5.3 minutes, which is approximately a 36% reduction compared to Membrane 1. Table 3 also shows that the MMD for each of the tested membranes did not vary significantly. This is in contrast to the differences in TOR observed for each membrane. Thus, in one embodiment, the spray time of the sprayer described herein is significantly reduced compared to prior art sprayers without affecting droplet size, as measured by MMD.

In addition to the membranes shown in Figures 5 and 6, membranes were also manufactured with a further reduced nozzle section and 3,000 through-holes 26 (Membranes 3 and 4, Table 3). In particular, Membrane 3 was laser drilled with a shorter nozzle section, while Membrane 4 was manufactured using a shorter nozzle section than Membrane 3. Table 3 shows that even with 3,000 holes (Membranes 3 and 4), the reduction in nozzle section length still resulted in an increase in TOR compared to Membrane 1, which had 6,000 holes. Comparison of Membranes 3 and 4 compared to Membrane 2 further shows that the combination of a higher number of holes (6,000 compared to 3,000) and a reduced nozzle section length increases the TOR of the sprayer.

In one embodiment, the use of a laser drilling process is advantageous compared to electroforming the through-holes. Compared to the funnel-shaped inlet and outlet of the electroformed through-holes, for example, as disclosed in WO 01/18280, the through-holes made by laser drilling shown in Figures 5 and 6 are substantially cylindrical or conical. When the through-holes are substantially cylindrical or conical compared to the funnel-shaped inlet and outlet of the electroformed through-holes, the vibrations of the membrane (i.e., its vibration speed) can be transmitted to the drug formulation over a larger area by friction. The drug formulation is then ejected from the outlet opening of the through-hole due to its own inertia, resulting in the collapse of the liquid jets to form an aerosol. Without wishing to be bound by theory, it is believed that because the electroformed membrane comprises a very curved surface with through-holes, the surface or area for transferring energy from the membrane to the liquid is reduced.

However, the present invention may also be implemented in electroformed films where the nozzle portion is defined by a continuous portion of the through-hole extending in a direction starting from the minimum diameter of the through-hole towards the first side until it reaches a diameter that is 2× or 3× the minimum diameter of the hole. In one embodiment, the total length of the through-hole is measured from the minimum diameter to the first side.

Referring again to FIG. 1 , to prevent the patient from having to remove or put down the therapeutic device from their mouth after inhaling the aerosol, the mouthpiece 3 has an opening 6 sealed by a resilient valve element 7 (exhalation valve). If the patient exhales into the mouthpiece 3 and thereby into the nebulizer chamber 2, the resilient valve element 7 opens so that the exhaled gas can escape from the interior of the therapeutic aerosol. Upon inhalation, ambient air flows through the nebulizer chamber 2. The nebulizer chamber 2 has an opening (not shown) sealed by another resilient valve element (inhalation valve). If the patient inhales through the mouthpiece 3 and inhales from the nebulizer chamber 2, the resilient valve element opens so that ambient air can enter the nebulizer chamber and mix with the aerosol and exit the interior of the nebulizer chamber 2 to be inhaled. A further description of this process is provided in U.S. Patent No. 6,962,151, which is incorporated by reference in its entirety for all purposes.

The nebulizer shown in FIG2 includes a cylindrical reservoir 10 for providing liquid for delivery to the membrane 5. As shown in FIG2, the oscillating membrane 5 can be arranged along the end wall 12 of the cylindrical liquid reservoir 10 to ensure that the liquid poured into the liquid reservoir directly contacts the membrane 5 when the aerosol generator is held in the position shown in FIG1. However, other methods can also be used to deliver the liquid to the oscillating membrane for the purpose of generating negative pressure in the liquid reservoir without making any changes to the design of the device according to the present invention.

The cylindrical liquid container 10 is open on the side facing the end wall 12. The opening is used to pour liquid into the liquid reservoir 10. A protrusion 15 is provided slightly below the opening on the outer surface 13 of the peripheral wall 14, which serves as a support when the liquid container is inserted into the appropriate receiving opening of the outer cover 35.

The open end of the liquid container 10 is closed by a flexible sealing member 16. The sealing member 16 is located 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-shaped manner, thereby forming a tapered continuous wall portion 17 within the sealing member 16 and being closed by a flat wall portion 18 of the sealing member 16. As discussed further below, force acts on the sealing member 16 via the flat wall portion 18, so in one embodiment, the flat wall portion 18 is thicker than other portions of the sealing member 16. The periphery of the flat wall portion 18 is spaced a certain distance from the tapered wall portion 17 so that when the flat wall portion 18 moves upward, the tapered wall portion 17 can be folded, as described with respect to FIG. 2 .

On the side of the flat wall portion 18 facing away from the interior of the liquid container, there is a projection comprising a truncated conical portion 19 and a cylindrical portion 20. Since the flexible material of the sealing element 16 allows the truncated conical portion 19 to deform, the above design enables the projection to be introduced and locked into the opening adapted to match the cylindrical portion.

In one embodiment, the aerosol generator 4 comprises a slidable sleeve 21 provided with an opening of this type, which is a substantially hollow, cylindrical body open on one side. The opening for the attachment of the sealing element 16 is contained in the end wall of the slidable sleeve 21. When the truncated cone 19 is locked in place, the end wall of the slidable sleeve 21 containing the opening rests on the flat wall portion 18 of the sealing element. The locking of the truncated cone 19 to the slidable sleeve causes a force to be transmitted from the slidable sleeve 21 to the flat wall portion 18 of the sealing element 16, causing the sealing portion 18 to follow the movement of the slidable sleeve 21 in the direction of the central longitudinal axis of the liquid container 10.

In a broad sense, the slidable sleeve 21 can be considered a slidable element, which can also be implemented, for example, as a slidable rod that can be snapped or inserted into a drilled hole. A characteristic of the slidable element 21 is that it can be used to apply a substantially linearly oriented force to the flat wall element 18 of the sealing element 16. In summary, a decisive factor in the operating mode of the aerosol generator according to the present invention is that the slidable element transmits a linear movement to the sealing element so that an increase in volume occurs within the liquid reservoir 10. This causes a negative pressure to be generated within the liquid reservoir 10, since the liquid reservoir 10 is otherwise airtight.

The sealing element 16 and the slidable element 21 can be produced in one piece, ie in one operation, but from different materials. Production techniques therefor are available in order to create a one-piece component of a nebulizer, for example in a fully automated production step.

In one embodiment, the slidable sleeve 21 is open to the truncated cone at the end facing the borehole, but at least two diametrically opposed lugs 22 and 23 protrude radially into the interior of the slidable sleeve 21. An annular shaft 24 surrounding the slidable sleeve extends radially outward. When the annular shaft 24 serves as a support for the slidable sleeve 21 in the position shown in FIG5 , the lugs 22 and 23 protruding into the interior of the slidable sleeve 21 serve 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 located on the outer peripheral wall of the rotating sleeve 26.

In one embodiment, the sprayer can be implemented with one of the protrusions 22 or 23 and one recess 25. In a further embodiment, two or more protrusions and a corresponding number of recesses are provided in an evenly distributed arrangement.

In one embodiment, the rotating sleeve 26 is also a cylinder with one side open, whereby the open end is arranged in the slidable sleeve 21 and thus faces the truncated cone 19, allowing the truncated cone 19 to pass through the rotating sleeve 26. In addition, the rotating sleeve 26 is arranged in the slidable sleeve 21 in such a manner that the protrusions 22 and 23 are located in the spiral groove 25. The inclination of the spiral groove 25 is designed so that when the rotating sleeve 26 rotates relative to the slidable sleeve 21, the protrusions 22 and 23 slide along the spiral groove 25, thereby causing a force parallel to the central longitudinal axis to be applied to the sliding protrusions 22 and 23 and thus to the slidable sleeve 21. This force displaces the slidable sleeve 21 in the direction of the central longitudinal axis, causing the sealing element 16, which is latched to the slidable sleeve bore by means of the truncated cone, to also be displaced substantially in a direction parallel to the central longitudinal axis.

The displacement of the sealing element 16 along the central longitudinal axis of the liquid container 10 generates a negative pressure in the liquid container 10, which is determined in particular by the distance by which the slidable sleeve 21 is displaced in the central longitudinal axis. The displacement causes the initial volume V _RI of the airtight liquid container 10 to increase to a volume V _RN , thereby generating a negative pressure. The displacement is further limited by the design of the spiral groove 25 in the rotating sleeve 26. In this way, the aerosol generator according to the present invention ensures that a negative pressure in the liquid reservoir 10 can be generated in the relevant area by means of simple structural measures.

In order to ensure that the forces for generating negative pressure when operating the device remain low, a rotating sleeve 26 and a handle 27 are integrally formed, the dimensions of which are selected so that the user can turn the handle 27 without having to exert great effort to manually turn the rotating sleeve 26. The handle 27 essentially has the shape of a flat cylinder or a truncated cone open on one side, so that a peripheral gripping area 28 is formed on the outer peripheral edge of the handle 27, which is touched by the user's hand to turn the handle 27.

Due to the design of the helical groove 25 and the relatively short distance that the slidable sleeve 21 moves in the longitudinal direction, which generates sufficient negative pressure, in one embodiment, a relatively small rotation angle is sufficient to rotate the handle 27 and thus the rotating sleeve 26. In one embodiment, the rotation angle is in the range of 45 to 360 degrees. This embodiment facilitates the operation of the device according to the present invention and the therapeutic aerosol generator equipped therewith.

In order to create a simple and uniformly operable unit 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 to support the slidable sleeve 21, which essentially comprises a flat cylinder with one side open. The diameter of the outer 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 aligned with the inner diameter of a cylindrical locking ring 31, which is provided concentrically with the gripping area 28 of the handle 27 but with a smaller diameter on the side of the handle 27 on which the rotating sleeve 26 is also arranged. The side of the cylindrical locking ring 31 facing the rotating sleeve includes a peripheral locking edge 32, which can engage with locking projections 33 located at intervals on the outer peripheral wall 30 of the bearing sleeve 29. This allows the handle 27 to be positioned on the bearing sleeve 29 , whereby, as shown in FIG. 5 , the handle 27 is placed at the open end of the bearing sleeve 29 and the latching edge 32 and the latching projection 33 latch with each other.

To secure the slidable sleeve 21, an opening is provided in the center of the closed end of the bearing sleeve 29, within which the slidable sleeve 21 is disposed, as can be seen in FIG2 . The annular shaft 24 of the slidable sleeve 21 is located on the end wall surface of the bearing sleeve 29, facing the handle, as shown in FIG2 . Two diametrically opposed projections 51 and 52 extend into the bearing opening, projecting into two longitudinal grooves 53 and 54 on the outer circumferential 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 lock for the slidable sleeve 21, ensuring that rotational movement of the rotating sleeve 26 does not result in rotation but rather in linear displacement of the slidable sleeve 21. As is apparent from FIG2 , this ensures that the slidable sleeve 21 is retained in the combination of the handle 27 and the bearing sleeve 29 in an axially displaceable manner, but is locked against rotation. If the handle 27 is rotated relative to the bearing sleeve 29, the rotating sleeve 26 is also rotated relative to the slidable sleeve 21, whereby the sliding protrusions 22 and 23 move along the spiral groove 25. This causes the slidable sleeve 21 to be axially displaced along the bearing sleeve 29 opening.

The guide projections 51 and 52 in the bearing opening and the longitudinal grooves 53 and 54 in the slidable sleeve 21 may not be required. In one embodiment, the guide projections 51 and 52 and the longitudinal grooves 53 and 54 are not present in the aerosol generator, and the truncated cone 19, the cylindrical portion 20 of the sealing element 16 and the large-area support of the slidable sleeve 21 for fixing the truncated cone to the flat sealing element portion 18 achieve an 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.

In one embodiment, an annular first sealing lip 34 is provided on the surface of the sealing end of the bearing sleeve 19 remote from the handle, concentric with the opening of the fixed slidable sleeve. The diameter of the first sealing lip 34 corresponds to the diameter of the outer peripheral wall 14 of the liquid container 10. As shown in FIG2 , this ensures that the first sealing lip 34 presses the sealing element 16 against the end of the outer peripheral wall of the liquid reservoir 10 in a manner that seals the liquid reservoir 10. In addition, the first sealing lip 34 also fixes 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 excessive force to ensure that the above-mentioned components of the device cannot rotate relative to each other.

In one embodiment, the required force is generated at least in part by the interaction between the handle 27 and the housing 35, in which the drug agent reservoir is integrally included or in which the drug agent (liquid) reservoir 10 is inserted, as shown in FIG2 . In this case, the drug agent reservoir 10 inserted in the sleeve having the peripheral protrusion 15 is located at the interval of the support 36 of the housing 35, which extends radially into the interior of the housing 35. This allows the liquid reservoir 10 to be easily removed from the housing 35 for cleaning purposes. In the embodiment shown in FIG2 , only the support is provided at the interval, thereby providing an opening for ambient air when the patient inhales, as described in more detail below.

2 , which is implemented on the one hand by the handle 27 and on the other hand by the housing 35. Shown on the housing 35 are locking projections 62 and 63. However, there are no special requirements regarding the design of the rotary lock, as long as the device according to the invention allows for the generation of negative pressure in the liquid reservoir 10.

In one embodiment, the liquid reservoir 10 is configured to have a volume V _RN of at least 16 mL, at least about 16 mL, at least 18 mL, at least about 18 mL, at least 20 mL, or at least about 20 mL, so that when, for example, an 8 mL amount of liquid (e.g., an aminoglycoside drug formulation) emitted in aerosol form is contained in (filled or injected into) the liquid reservoir 10, an air cushion of 8 mL or about 8 mL is provided. That is, the ratio of the volume V _RN within the liquid reservoir 10 to the initial volume V _L of the liquid is at least 2.0 and the ratio between the volume _VA of the gas and the V _L of the liquid is at least 1.0. Liquid reservoirs having volumes V _RN of about 15.5 mL, about 19.5 mL, and about 22.5 mL have been shown to be effective, and their efficiency increases with increasing V _RN .

In one embodiment, the ratio between _VRN and _VL is at least 2.0, at least about 2.0, at least 2.4, at least about 2.4, at least 2.8, or at least about 2.8. In one embodiment, the ratio between _VA and _VL is at least 1.0, at least 1.2, at least 1.4, at least 1.6, or at least 1.8. In another embodiment, the ratio between _VA and _VL 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.

In one embodiment, the volume of the air cushion is at least 2 mL, at least about 2 mL, at least 4 mL, at least about 4 mL, at least 6 mL, at least about 6 mL, at least 8 mL, at least about 8 mL, at least 10 mL, at least about 10 mL, at least 11 mL, at least about 11 mL, at least 12 mL, at least about 12 mL, at least 13 mL, at least about 13 mL, at least 14 mL, or at least about 14 mL. In one embodiment, the volume of the air cushion is at least about 11 mL or at least about 14 mL. In one embodiment, the volume of the air cushion is from about 6 mL to about 15 mL, and the ratio between V _RN and V _L is from at least about 2.0 to at least about 3.0. In further embodiments, the ratio between V _RN and V _L is from at least about 2.0 to at least about 2.8.

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

In one embodiment, the ratio of volume V _RN to the initial volume V _L of the liquid is at least 2.0. Theoretically, the increasing volume V _RN of the liquid reservoir 10, which is infinitely enlarged, will produce a nearly stable negative pressure range. In one embodiment, the ratio of volume V _RN to the initial volume V _L of the liquid is in the range of 2.0 to 4.0, and in further embodiments, 2.4 to 3.2. Two examples of the ratio range (V _RN /V _L ) for the initial volume V _L of different liquids ranging from 4 mL to 8 mL are shown in Table 4 below.

The system provided herein can be used to treat a variety of lung infections in a subject in need thereof. Available lung infections (e.g., cystic fibrosis patients) that can be treated by the methods of the present invention include gram-negative infections. In one embodiment, the infection caused by the following bacteria can be treated with the system and preparation provided herein: Pseudomonas (e.g., Pseudomonas aeruginosa, Pseudomonas paucimobilis, Pseudomonas putida, Pseudomonas fluorescens, and Pseudomonas acidovorans), Burkholderia (e.g., Burkholderia pseudomallei, Burkholderia cepacia, Burkholderia cepacia complex, Burkholderia reluctance, Burkholderia fungi, Burkholderia gladiolus, Burkholderia multivora, Burkholderia vietnamese, Burkholderia pseudomallei), Burkholderia spp. Burkholderia, Burkholderia aphida, Burkholderia anthina, Burkholderia brasiliensis, Burkholderia californica, Burkholderia caryophyllus, Burkholderia dianthus), Staphylococci (e.g., Staphylococcus aureus, Staphylococcus auris, Staphylococcus carnosus, Staphylococcus epidermidis, Staphylococcus lyonii), methicillin-resistant Staphylococcus aureus (MRSA), Streptococci (e.g., Streptococcus pneumoniae), Escherichia coli, Klebsiella, Enterobacter, Serratia, Haemophilus, Yersinia pestis, Mycobacterium, nontuberculous mycobacteria (e.g., Mycobacterium avium, Mycobacterium avium hominis subsp. suis (MAH), Mycobacterium abscessus, Mycobacterium chelonae, Mycobacterium bovis, Mycobacterium kansasii, Mycobacterium ulcerans, Mycobacterium avium, Mycobacterium avium complex (MAC) (Mycobacterium avium and Mycobacterium intracellulare), Mycobacterium spp., Mycobacterium kansasii, Mycobacterium exogenum, Mycobacterium immunogenicum, Mycobacterium toad, Mycobacterium marinum, Mycobacterium malmoe, Mycobacterium ulcerans, Mycobacterium avium complex (MAC) (Mycobacterium avium and Mycobacterium intracellulare), Mycobacterium spp. Mycobacterium marinum, Mycobacterium mucogenes, Mycobacterium non-chromogenic, Mycobacterium scrofula, Mycobacterium simiae, Mycobacterium smegmatis, Mycobacterium surga, Mycobacterium terrestris, Mycobacterium terrestris complex, Mycobacterium hemophilus, Mycobacterium geneva, Mycobacterium asiatica, Mycobacterium subspecies, Mycobacterium gordonii, Mycobacterium non-chromogenic, Mycobacterium tristis, Mycobacterium lentiginosa, Mycobacterium occulta, Mycobacterium fortuitum, Mycobacterium fortuitum complex (Mycobacterium fortuitum and Mycobacterium chelonae)).

In one embodiment, the systems described herein are used to treat infections caused by nontuberculous mycobacteria. In one embodiment, the systems described herein are used to treat infections caused by Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium, or Mycobacterium avium complex. In a further embodiment, one or more systems described herein are used to treat infections caused by Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium, or Mycobacterium avium complex in patients with cystic fibrosis. In yet further embodiments, the Mycobacterium avium infection is Mycobacterium avium subspecies hominissuis.

In one embodiment, a system provided herein is used to treat a lung infection in a cystic fibrosis patient. In a further embodiment, the lung infection is a Pseudomonas infection. In yet a further embodiment, the Pseudomonas infection is Pseudomonas aeruginosa. In a further embodiment, the aminoglycoside in the system is amikacin.

In one embodiment, the system provided herein is used to treat or prevent Pseudomonas aeruginosa, Mycobacterium abscessus, Mycobacterium avium or Mycobacterium avium complex lung infections in patients with cystic fibrosis or non-cystic fibrosis. In a further embodiment, the system provided herein comprises a liposomal aminoglycoside formulation. In a further embodiment, the aminoglycoside is selected from amikacin, apramycin, arbekacin, astamicin, capreomycin, dibekacin, neomycin B, gentamicin, hygromycin B, isopamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribosomycin, sisomicin, spectinomycin, streptomycin, tobramycin, methyl zithromycin or a combination thereof. In yet a further embodiment, the aminoglycoside is amikacin, for example, amikacin sulfate.

An obstacle to treating infectious diseases such as Pseudomonas aeruginosa (a major cause of chronic disease in patients with cystic fibrosis) is drug penetration within the saliva/biofilm barrier on epithelial cells (Figure 7). In Figure 7, the circular diagram represents liposomes/complexed aminoglycosides, the "+" symbol represents free aminoglycoside, the "-" symbol represents mucin, alginate, and DNA, and the solid bar symbol represents Pseudomonas aeruginosa. This barrier contains colonized and planktonic Pseudomonas aeruginosa embedded in alginate or exopolysaccharides from the bacteria, as well as DNA from damaged white blood cells and mucin from lung epithelial cells, all of which have a net negative charge. The negative charge combines and prevents positively charged drugs such as aminoglycosides from penetrating, rendering them biologically ineffective (Mendelman et al., 1985). Without wishing to be bound by theory, encapsulation of aminoglycosides within liposomes or lipoplexes shields or partially shields nonspecific binding of aminoglycosides to saliva/biofilms, allowing the liposomes or lipoplexes (with encapsulated aminoglycosides) to penetrate (Figure 7).

In another embodiment, a system provided herein is used to treat a non-tuberculous mycobacterial lung infection in a patient. In a further embodiment, a system provided herein comprises a liposomal amikacin formulation.

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.

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.

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

As provided herein, the present invention provides aminoglycoside formulations for administration by inhalation. In one embodiment, the MMAD of the aerosol is from about 3.2 μm to about 4.2 μm, as measured by an Anderson Cascade Impactor (ACI), or from about 4.4 μm to about 4.9 μm, as measured by a new generation impactor (NGI).

In one embodiment, the nebulization time of an effective amount of an 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 an effective amount of an aminoglycoside formulation provided herein is less than 15 minutes or less than 13 minutes. In one embodiment, the nebulization time of an effective amount of an aminoglycoside formulation provided herein is about 13 minutes.

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

Example

The present invention is further illustrated by reference to the following examples. However, it should be noted that these examples, like the above-described embodiments, are illustrative and should not be construed as limiting the scope of the present invention in any way.

Example 1: Comparison of nebulizer reservoir volumes

In this example, the aerosol generators were research nebulizers from Pari Pharma GmbH, Germany, modified for use with the liposomal aminoglycoside formulations provided herein. The first aerosol generator had an initial liquid reservoir volume V RI of 13 mL (A), the second had an initial volume V _RI of 17 mL (B), the third had an initial volume V RI of 22 mL (C), and the fourth had an initial volume V RI of 20 mL (D). Thus, the initial volume V _RN of the first aerosol generator was 15.5 mL, the second had an initial volume V RI of 19.5 mL, the third had an initial volume V RI of 24.5 mL, and the fourth had an initial volume V RI of 22.5 mL.

8 mL of liposomal amikacin formulation was injected into the liquid reservoir 10. As shown in FIG8 , after the 8 mL formulation was completely discharged into the liquid reservoir, the 8 mL air cushion resulted in an aerosol generation time period of 14 to 16 minutes. 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 ( FIG6 ).

In addition, the first (A) and third (C) versions of the aerosol generator were used with 8 mL of liposomal amikacin formulation. An initial negative pressure of 50 mbar or less was generated within the liquid reservoir. In addition, the negative pressure during the aerosol generation process was measured and displayed in FIG9 along with the aerosol generation time. In other words, FIG9 shows experimental data comparing the negative pressure range during the aerosol generation time for a liquid reservoir (C) with a V _RN volume of 24.5 mL and a liquid reservoir (A) with a V _RN volume of 15.5 mL. The initial amount V _L of the amikacin formulation was 8 mL and the initial negative pressure was approximately 50 mbar. The figure shows that the larger air cushion prevents the negative pressure from increasing beyond the critical value of 300 mbar.

The above-described nebulizer was used to measure the dependence of aerosol generator efficiency (proportional to the liquid output rate or total output rate) on different negative pressures. The experiments used liposomal amikacin formulations with viscosities ranging from 5.5 to 14.5 mPa×s at shear forces of 1.1 to 7.4 Pa (thixotrope). As shown in Figure 10 , efficiency was optimal within the negative pressure range of 150 mbar to 300 mbar. Also shown in Figure 10 , efficiency decreased at negative pressures below approximately 150 mbar and above 300 mbar.

In addition, the same liposomal amikacin formulation as in FIG8 was used in four different aerosol generators based on the modified VIZOL®, wherein the first aerosol generator (A) was a modified VIZOL® with an increased volume V _RN of a liquid reservoir of 19.5 mL and was filled with 8 mL of the liposomal amikacin formulation.

The second aerosol generator (B) had a 16 mL reservoir with an increased volume V _RN containing 8 mL of the liposomal amikacin formulation, and the third aerosol generator (C) had a 24.5 mL reservoir with an increased volume V _RN containing 8 mL of the liquid. The fourth aerosol generator had a 22.5 mL liquid reservoir with an increased volume V _RN and was filled with 8 mL of the liposomal amikacin formulation.

Figure 11 shows experimental data from these four aerosol generators, each loaded with 8 mL of a liposomal amikacin formulation. The results show that the aerosol generation time required for complete release of the liposomal amikacin formulation from the liquid reservoir is related to the ratio of the increased volume of the liquid reservoir (V _RN ) to the initial volume of liquid in the liquid reservoir before use (V _L ). Figure 11 demonstrates that the improved aerosol generator device (A) requires approximately 16 minutes of aerosol generation time, and that as the V _RN /V _L ratio increases, the aerosol generation time decreases. The data also demonstrate that the aerosol generation time of the third aerosol generator device (C) can be reduced by approximately 4 minutes to less than 12 minutes.

Thus, the data provided in Example 1 demonstrate that a larger air cushion allows the aerosol generator to operate in an efficient negative pressure range for a longer period of time, thereby significantly reducing the total aerosol generation time. Thus, even large amounts of liquid, such as 8 mL, can be nebulized (released as an aerosol) in a period of less than 12 minutes.

Example 2: Aerosol Properties of Amikacin Formulations

Eleven different batches of liposomal amikacin formulations were examined using a modified nebulizer (i.e., modified for the liposomal aminoglycoside formulations described herein) having a modified 40-mesh membrane and a reservoir with an 8 mL liquid capacity and the aforementioned air cushion assembled as described herein. Cascade impaction was performed using either an ACI (Anderson Cascade Impactor) or an NGI (New Generation Impactor) to determine aerosol properties: total aerodynamic median diameter (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF).

Measurement of total median aerodynamic diameter (MMAD) using ACI

Anderson cascade impactor (ACI) is used for MMAD measurement and spray operation is carried out in ClimateZone room (WestechInstruments Inc., GA) to maintain temperature and relative humidity % during spraying. ClimateZone is pre-set to a temperature of 18°C and a relative humidity of 50%. ACI is assembled and loaded into ClimateZone interior. A probe thermometer (VWR dual thermometer) is connected to the surface of ACI at stage 3 to monitor the temperature of ACI. When the temperature of ACI reaches 18±0.5°C, spraying is started.

Since the 8 mL handle (handset) was loaded with 8 mL, it was found that the ACI could not process the entire 8 mL dose; that is, the liposomal amikacin formulation deposited on ACI plate 3 overflowed. It was determined that the percentage of drug distribution on each ACI stage was not affected by the amount of liposomal amikacin formulation collected within the ACI as long as no liquid overflowed in the ACI stage 3 (data not shown). Therefore, for nebulization, the nebulizer was loaded with 4 mL of liposomal amikacin formulation and nebulized until empty, or loaded with 8 mL of liposomal amikacin formulation and nebulized for a collection time of approximately 6 minutes (i.e., ~4 mL).

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

After collecting the spray, remove ACI collection plates 0, 1, 2, 3, 4, 5, 6 and 7, and each is placed in its own culture dish. Add an appropriate amount of extract (plate 2, 3 and 4 is 20 mL, plate 0, 1, 5, 6 and 7 is 10 mL) to dissolve the preparation deposited on each plate in each culture dish. Use mobile phase C to further appropriately dilute the sample of plate 0, 1, 2, 3, 4, 5 and 6 for HPLC analysis. The sample of plate 7 is directly analyzed by HPLC without any further dilution. The ACI filter is also transferred to a 20 mL bottle and 10 mL of extract is added. The capped bottle is vortexed to dissolve any preparation deposited thereon. The liquid sample from the bottle is filtered (0.2 μm) into the HPLC bottle for HPLC analysis. The introduction port of the connector is also rinsed with 10 mL of extract to dissolve the preparation deposited thereon, and the collected sample is diluted and analyzed by HPLC with 2 times. Based on the amount of amikacin deposited on each stage of the impactor, the total median aerodynamic diameter (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF) were calculated.

With the nebulizer loaded with 8 mL and nebulized for 6 minutes, the fine particle dose (FPD) was normalized to the formulation spray volume to compare the FPD in all experiments. The FPD (normalized to the formulation spray volume) was calculated according to the following formula:

Measurement of total median aerodynamic diameter (MMAD) using NGI

A new generation impactor (NGI) was also used for MMAD measurements and spraying was performed in a ClimateZone chamber (Westech Instruments Inc., GA) to maintain temperature and RH% during the spraying process. The ClimateZone was pre-set to a temperature of 18°C and a relative humidity of 50%. The NGI was assembled and loaded into the ClimateZone. A probe thermometer (VWR dual thermometer) was attached to the surface of the NGI to monitor the temperature of the NGI. When the temperature of the NGI reached 18 ± 0.5°C, spraying was started.

8 mL of liposomal amikacin formulation was added to the nebulizer and sprayed. When aerosol was no longer observed, the timer was stopped. The spray was collected at a flow rate of 15 L/min in an NGI cooled to 18°C. The spray time was recorded and the spray rate was calculated based on the weight difference (spray amount) divided by the time interval.

After aerosol collection is complete, the NGI tray with the tray support is removed from the NGI. An appropriate amount of extraction solution is added to NGI cups 1, 2, 3, 4, 5, 6, 7, and the MOC to dissolve the formulation deposited on these cups. This material is transferred to a volumetric flask. For NGI cups 1, 2, and 6, a 25ml volumetric flask is used; for NGI cups 2, 3, and 4, a 50ml volumetric flask is used. More extraction solution is added to the cup and transferred to the volumetric flask again. This step is repeated several times to completely transfer the formulation deposited on the NGI cup to the volumetric flask. The volumetric flask is filled to a final volume of 25ml or 50ml and shaken thoroughly before sampling. Samples from cups 1, 2, 3, 4, 5, 6, and 7 are further diluted appropriately with mobile phase C for HPLC analysis. The sample from the MOC is analyzed directly by HPLC without any further dilution. The NGI filter is also transferred to a 20ml vial and 10ml of extraction solution is added. The capped vial is vortexed to dissolve any formulation deposited on it. The liquid sample from the vial was filtered (0.2 micron) into an HPLC vial for HPLC analysis. The introduction port with the connector was also rinsed with 10 mL of the extraction solution to dissolve the preparation deposited thereon, and the sample was collected and analyzed by HPLC using an 11-fold dilution.

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

The FPD was normalized to the formulation spray volume in order to compare the FPD in all experiments. The FPD (normalized to the formulation spray volume) was calculated according to the following formula:

The results of these experiments are provided in Figures 12 and 13 and Table 5 below.

Example 3: Spray Rate Study

Spray rate study (grams of preparation sprayed per minute) was carried out in a biological safety cabinet (model 1168, B2 type, FORMA Scientific). First, the empty assembled sprayer (handle and aerosol head with a mouthpiece) (W ₁ ) was weighed, then a certain volume of preparation was added, and the sprayer device was weighed again (W ₂ ). The sprayer and timer were started, and the spray preparation was collected in a freezing impactor at a flow rate of ~8L/min (see Figure 14 for details of the experimental scheme). When no aerosol was observed, the timer was stopped. The sprayer was weighed again (W ₃ ), and the spray time (t) was recorded. The total spray preparation was calculated as W ₂ -W ₃ and the total drug residue after spraying was calculated as W ₃ -W ₁ . The spray rate of the preparation was calculated according to the following formula:

For liposome amikacin nebulized using a nebulizer assembled according to the instructions (24 aerosol heads were selected and used in these studies), the nebulization rate (in g/min) and other relevant results are obtained in Table 6.

Example 4: Percentage of Amikacin Incorporated After Nebulization and Characterization of the Nebulizer

Measurement of free amikacin and liposomal complexed amikacin in the spray of Example 3. As described in Example 3, the spray was collected in a cryoimpinger at a flow rate of 8 L/min (Figure 14).

The spray 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 remove any formulation deposited in the impinger and transfer it to the flask. To measure the free amikacin concentration in the spray, 0.5 mL of the diluted spray from the volumetric flask was loaded onto an Ultra-0.5 mL 30K centrifugal filter device (regenerated cellulose, 30K MWCO, Millipore) and centrifuged at 5000 g for 15 minutes at 15°C. An appropriate amount of the filtrate was diluted 51-fold with mobile phase C solution. The amikacin concentration was measured by HPLC. To measure the total amikacin concentration in the spray, an appropriate amount of the diluted spray from the volumetric flask was diluted (and dissolved) 101-fold in an extraction solution (perfluoropentanoic acid: 1-propanol: water (25:225:250, v/v/v)), and the amikacin concentration was measured by HPLC.

The percentage of amikacin bound after spraying was calculated by the following formula:

The percentage of amikacin incorporated 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.

During this study, the total amikacin concentration in the liposomal amikacin formulation was measured with the remaining samples using the same HPLC and amikacin standard. The value obtained was 64 mg/mL amikacin. The % amikacin bound after nebulization ranged from 58.1% to 72.7%, with a mean of 65.5 ± 2.6%. The total amount of amikacin recovered for 8 mL of nebulized liposomal amikacin formulation ranged from 426 mg to 519 mg, with a mean of 476 ± 17 mg. The calculated amount of amikacin nebulized (based on the weight of the nebulized liposomal amikacin formulation in Table 7) ranged from 471 mg to 501 mg, with a mean of 490 ± 8 mg. The total amikacin recovery ranged from 91% to 104%, with a mean of 97 ± 3% (n = 72).

Liposome size

Liposomal amikacin formulations (64 mg/mL amikacin) (before or after nebulization) were appropriately diluted with 1.5% NaCl and liposome particle size was measured by light scattering using a Nicomp 380 submicron particle sizer (Nicomp, Santa Barbara, CA).

The liposome size after nebulization of the liposomal amikacin formulation sprayed through a 24-spray aerosol head with an 8 mL reservoir handle was measured. The size of the liposomes ranged from 248.9 nm to 288.6 nm, with an average of 264.8 ± 6.7 nm (n = 72). These results are provided in Table 8. The average liposome diameter before nebulization was approximately 285 nm (284.5 nm ± 6.3 nm).

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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.

The embodiments illustrated and discussed in this specification are intended only to teach those skilled in the art the best ways known to the inventors to make and use the present invention. The above embodiments of the present invention may be modified and varied without departing from the present invention, as will be appreciated by those skilled in the art in light of the foregoing teachings. It is therefore understood that, within the scope of the claims and their equivalents, the present invention may be practiced otherwise than as specifically described.

Claims (27)

1. A system for treating a patient's lung infection, comprising: (a) A pharmaceutical formulation comprising an aqueous dispersion of a liposome-complexed aminoglycoside, wherein the lipid component of the liposome-complexed aminoglycoside is present at 45-60 mg/mL and consists of dipalmitoylphosphatidylcholine (DPPC) and cholesterol, and (b) A sprayer comprising a vibrating mesh component, the vibrating mesh component including a plurality of through holes, each having a nozzle portion, wherein the length of the nozzle portion is less than 25 μm and the diameter is the smallest diameter of the through hole, and wherein the nozzle portion forms a boundary with a larger diameter of the through hole, the larger diameter of the through hole being 1.5 times, 2 times, or 3 times the smallest diameter of the through hole, wherein the ratio of the total length of the through holes in the extending direction to the length of the nozzle portion of the through holes in the extending direction is at least 4. The sprayer produces an aerosol of the pharmaceutical formulation at a rate of 0.60 g per minute to 0.80 g per minute, wherein the fine particulate fraction (FPF) of the aerosol is greater than or equal to 64% as measured by an Anderson cascade collisionor (ACI) or greater than or equal to 51% as measured by a next-generation collisionor (NGI), and the percentage of bound aminoglycosides in the aerosol is 60% to 70%. 2. The system of claim 1, wherein the aminoglycoside is selected from amikacin, apramycin, abekacin, asmicin, capreomycin, dibekacin, neomycin B, gentamicin, hygromycin B, isopamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodestreptomycin, ribosomycin, sisomicin, spectinomycin, streptomycin, tobramycin, methylsodium thiomycin, or combinations thereof. 3. The system of claim 1, wherein the aminoglycoside is amikacin. 4. The system of claim 3, wherein the aminoglycoside is amikacin sulfate. 5. The system of claim 1, wherein the liposomes of the aminoglycoside complex are monolayer liposomes, multilayer liposomes, or a mixture thereof. 6. The system of claim 1, wherein the aminoglycoside is amikacin, and the liposomes of the aminoglycoside complex are monolayer liposomes, multilayer liposomes, or a mixture thereof. 7. The system of claim 1, wherein the fine particulate fraction (FPF) of the atomized pharmaceutical formulation is 64% to 80% as measured by ACI; or 51% to 65% as measured by NGI. 8. Use of a pharmaceutical formulation comprising an aqueous dispersion of an aminoglycoside containing a liposome complex in the preparation of a medicament for treating a patient's lung infection, wherein the lipid component of the aminoglycoside containing the liposome complex is present at 45-60 mg/mL and is composed of dipalmitoylphosphatidylcholine (DPPC) and cholesterol, the use comprising: The pharmaceutical preparation is atomized by spraying it at a rate of 0.60 g/min to 0.80 g/min using a sprayer, wherein the sprayer includes a vibrating mesh component comprising a plurality of through-holes, each having a nozzle portion, wherein the nozzle portion has a length of less than 25 μm and a diameter equal to the smallest diameter of the through-hole, and wherein the nozzle portion forms a boundary with a larger diameter of the through-hole, the larger diameter of the through-hole being 1.5 times, 2 times, or 3 times the smallest diameter of the through-hole, wherein the ratio of the total length of the through-holes in the extending direction to the length of the nozzle portion of the through-holes in the extending direction is at least 4; and The aerosolized drug preparation is administered into the patient's lungs; The aerosolized pharmaceutical formulation comprises a mixture of free aminoglycosides and liposome-complexed aminoglycosides, wherein the fine particulate fraction (FPF) of the aerosol is greater than or equal to 64% as measured by an Anderson cascade collisionor (ACI) or greater than or equal to 51% as measured by a next-generation collisionor (NGI), and the percentage of bound aminoglycosides in the aerosol is 60% to 70%. 9. The use of claim 8, wherein the aminoglycoside is selected from amikacin, apramycin, abekacin, asmicin, capreomycin, dibekacin, neomycin B, gentamicin, hygromycin B, isopamicin, kanamycin, neomycin, netilmicin, paromomycin, ribosomycin, sisomicin, rhodestreptomycin, spectinomycin, streptomycin, tobramycin, methylsodium thiosulfate, or combinations thereof. 10. The use of claim 8, wherein the aerosolized pharmaceutical formulation is administered once daily during a single dosing period. 11. The use of claim 8, wherein the aminoglycoside is amikacin. 12. The use of claim 11, wherein the aminoglycoside is amikacin sulfate. 13. The use of claim 8, wherein the patient has cystic fibrosis. 14. The use of claim 8, wherein the lung infection is a nontuberculous mycobacterial lung infection. 15. The use of claim 8, wherein the fine particulate fraction (FPF) of the atomized pharmaceutical formulation is 64% to 80% as measured by an Anderson cascade collisioner (ACI); or 51% to 65% as measured by a next-generation collisioner (NGI). 16. The use of claim 14, wherein the nontuberculous mycobacterial pulmonary infection is Mycobacterium avium, Mycobacterium avium human-swine subspecies (MAH), Mycobacterium abscessus, Mycobacterium chelonae, Mycobacterium bolletii, Mycobacterium kansasii, Mycobacterium ulcerans, Mycobacterium avium complex (MAC), Mycobacterium conspicuum, Mycobacterium peregrinum, or immunogenic mycobacteria. Mycobacterium immunogenum, Mycobacterium xenopi, Mycobacterium marinum, Mycobacterium malmoense, Mycobacterium mucogenicum, Mycobacterium nonchromogenicum, Mycobacterium scrofulaceum, Mycobacterium simiae, Mycobacterium smegmatis, Mycobacterium szulgai, Mycobacterium terrae, Mycobacterium terrae complex Lung infections caused by mycobacteria including mycobacterium haemophilum, mycobacterium genavense, mycobacterium asiaticum, mycobacterium shimoidei, mycobacterium gordonae, mycobacterium triplex, mycobacterium lentiflavum, mycobacterium celatum, mycobacterium fortuitum, or mycobacterium fortuitum complex. 17. The use of claim 16, wherein the nontuberculous mycobacterial lung infection is a Mycobacterium avium complex (MAC) lung infection. 18. The use of claim 17, wherein the aminoglycoside is amikacin. 19. The use of claim 18, wherein the aminoglycoside is amikacin sulfate. 20. The use of claim 8, wherein the lung infection is a nontuberculous mycobacterial lung infection, and the aminoglycoside is amikacin sulfate. 21. The use of claim 8, wherein the lung infection is Mycobacterium avium complex (MAC) lung infection, and the aminoglycoside is amikacin sulfate. 22. The system of claim 1, wherein the total gas dynamic median diameter (MMAD) of the aerosol is less than 4.2 μm as measured by an Anderson cascade collisioner (ACI) or less than 4.9 μm as measured by a next-generation collisioner (NGI). 23. The system of claim 22, wherein the total gas dynamic median diameter (MMAD) of the aerosol is measured to be 3.2 μm to 4.2 μm by an Anderson cascade collisioner (ACI); or 4.4 μm to 4.9 μm by a next-generation collisioner (NGI). 24. The use of claim 8, wherein the total gas dynamic median diameter (MMAD) of the aerosol is less than 4.2 μm as measured by an Anderson cascade collisioner (ACI) or less than 4.9 μm as measured by a next-generation collisioner (NGI). 25. The use of claim 24, wherein the total gas dynamic median diameter (MMAD) of the aerosol is measured to be 3.2 μm to 4.2 μm by an Anderson cascade collisioner (ACI); or 4.4 μm to 4.9 μm by a next-generation collisioner (NGI). 26. The system of claim 1, wherein the DPPC is present at 30-40 mg/mL and the cholesterol is present at 15-20 mg/mL. 27. The use of claim 8, wherein the DPPC is present at 30-40 mg/mL and the cholesterol is present at 15-20 mg/mL.