Compositions of clofazimine, combinations comprising the same, processes for its preparation, uses and methods comprising the same

Abstract

To provide a pharmaceutical composition for inhalation comprising highly hydrophobic clofazimine.SOLUTION: The present invention relates to a pharmaceutical composition for inhalation comprising a therapeutically effective dose of clofazimine, wherein the clofazimine is provided in the form of a suspension, and to a process for preparing the same. Furthermore, the present invention provides a pharmaceutical combination comprising clofazimine in the form of an aerosol for pulmonary inhalation. The combinations and compositions provided by the present invention can be used in the treatment and / or prophylaxis of pulmonary infections caused by mycobacteria and other gram-positive bacteria, as well as pulmonary fungal infections.SELECTED DRAWING: None

Description

Technical Field

[0001] This application claims the benefit of U.S. Provisional Application No. 62 / 722,048, filed Aug. 23, 2018, and this application also claims the benefit of U.S. Provisional Application No. 62 / 796,322, filed Jan. 25, 2019, the contents of both of which are hereby incorporated by reference herein as part of this specification.

[0002] The present invention relates to an inhalation pharmaceutical composition comprising a therapeutically effective dose of clofazimine, wherein the clofazimine is provided in the form of a suspension; a process for preparing the same; and uses and treatment methods comprising the same. Further, the present invention provides a combination medicament comprising clofazimine in the form of an aerosol for transpulmonary inhalation.

[0003] The combinations and compositions provided by the present invention can be used for the treatment and / or prevention of lung infections and pulmonary fungal infections caused by mycobacteria and other Gram-positive bacteria.

Background Art

[0004] Clofazimine is a highly hydrophobic (LogP = 7.66) antibiotic, namely, lofepramine, which has antibacterial and anti-inflammatory activities and was first described in 1957. The structural formula is shown below.

Chemical Formula

[0005] The exact mechanism by which clofazimine exerts its antibacterial action is not known. However, By preferentially binding to the DNA of mycobacteria, it is known to inhibit DNA replication and cell proliferation. Other proposed mechanisms of action include membrane damage / destabilization, generation of lysophospholipids that destabilize the membrane, potassium transport, and / or disruption of the intracellular redox cycle. Chlofazimine shows excellent activity against Mycobacterium tuberculosis (MTB), including multidrug-resistant strains, in vitro. However, until recently, it was generally considered ineffective for the treatment of pulmonary tuberculosis (e.g., Cholo M et al., J Antimicrob Chemother, 2012 Feb, 67(2):290 (Mycobacterium tuberculosis)(MTB) while until recently it was generally considered ineffective for the treatment of pulmonary tuberculosis (e.g., Cholo M et al., J Antimicrob Chemother, 2012 Feb, 67(2):290 - 8). Antimicrob Chemother, 2012 Feb, 67(2):290 - 8). Antimicrob Chemother, 2012 Feb, 67(2):290 - 8). -8).

[0006] Chlofazimine is one of the three main drugs recommended by the World Health Organization (WHO) for the treatment of leprosy caused by Mycobacterium leprae. In recent years, it has been increasingly used in the treatment of other mycobacterial infections such as drug-resistant tuberculosis caused by non-tuberculous mycobacteria (NTM). Chlofazimine is one of the three main drugs recommended by the World Health Organization (WHO) for the treatment of leprosy caused by Mycobacterium leprae. In recent years, it has been increasingly used in the treatment of other mycobacterial infections such as drug-resistant tuberculosis caused by non-tuberculous mycobacteria (NTM). (World Health Organization) for the treatment of leprosy caused by Mycobacterium leprae. In recent years, it has been increasingly used in the treatment of other mycobacterial infections such as drug-resistant tuberculosis caused by non-tuberculous mycobacteria (NTM). Chlofazimine is one of the three main drugs recommended by the World Health Organization (WHO) for the treatment of leprosy caused by Mycobacterium leprae. In recent years, it has been increasingly used in the treatment of other mycobacterial infections such as drug-resistant tuberculosis caused by non-tuberculous mycobacteria (NTM). Chlofazimine is poorly soluble in water and shows high membrane permeability, so it is classified as a class II drug in the Biopharmaceutics Classification System (BCS).

[0007] Chlofazimine is poorly soluble in water and shows high membrane permeability, so it is classified as a class II drug in the Biopharmaceutics Classification System (BCS). Chlofazimine is poorly soluble in water and shows high membrane permeability, so it is classified as a class II drug in the Biopharmaceutics Classification System (BCS). To overcome the problems associated with low oral absorption and low bioavailability of the drug.

[0008] [[ID=4ID=41]]To overcome the problems associated with low oral absorption and low bioavailability of the drug. For the purpose of optimizing oral dosage forms, various strategies such as micronization, nanoparticle formation (nanonization), recrystallization using supercritical fluids (supercritical fluid re-crystallization), spray freeze drying into liquid, solid dispersions, and solid solutions have been used. zation), recrystallization using supercritical fluids (supercritical fluid re-crystallization), spray freeze drying into liquid (spray fr eeze drying into liquid), solid dispersions, and solid solutions have been used. Such strategies have been used.

[0009] Since clofazimine is classified as a BCS class II drug, it is usually considered an ideal option to formulate it in a solid dispersion to improve oral bioavailability (see, for example, Bhusnure et al. IJRPC 2014, 4 (4), 906-918). Accordingly, lipophilic clofazimine is usually administered as a microcrystalline suspension in an oil-wax base for the purpose of improving oral absorption. There is a significant difference in the absorption rate by the human body after oral administration (45-62%). The adverse effects of clofazimine are dose-related and mainly affect the skin, eyes, and gastrointestinal tract. The QT prolongation side effect is accompanied by the occurrence of reddish-brown discoloration of the skin and conjunctiva and shows reversibility, slowly recovering upon discontinuation of the drug. This is due to its long-term systemic retention. (4), 906-918).

[0010] Accordingly, lipophilic clofazimine is usually administered as a microcrystalline suspension in an oil-wax base for the purpose of improving oral absorption. The QT prolongation side effect is accompanied by the occurrence of reddish-brown discoloration of the skin and conjunctiva and shows reversibility, slowly recovering upon discontinuation of the drug. This is due to its long-term systemic retention. The absorption rate by the human body after oral administration varies significantly (45-62%). The adverse effects of clofazimine are dose-related and mainly affect the skin, eyes, and gastrointestinal tract. The QT prolongation side effect is accompanied by the occurrence of reddish-brown discoloration of the skin and conjunctiva and shows reversibility, slowly recovering upon discontinuation of the drug. This is due to its long-term systemic retention. The QT prolongation side effect is accompanied by the occurrence of reddish-brown discoloration of the skin and conjunctiva and shows reversibility, slowly recovering upon discontinuation of the drug. This is due to its long-term systemic retention. This is due to its long-term systemic retention.

[0011] Mycobacterium is a genus of Actinobacteria and an independent family called Mycobacteriaceae. Mycobacteria have a rod-like shape and a waxy outer membrane. Mycobacterium is a genus of Actinobacteria and an independent family called Mycobacteriaceae. Mycobacteria have a rod-like shape and a waxy outer membrane. Mycobacteria have a rod-like shape and a waxy outer membrane.

[0012] As such, mycobacteria can be divided into three groups: · Mycobacterium tuberculosis group - the pathogen causing tuberculosis · Mycobacterium leprae group - the pathogen causing leprosy · Nontuberculous mycobacteria (NTM) - including all mycobacteria other than Mycobacterium tuberculosis and Mycobacterium leprae. Mycobacterium abscessus complex (MABSC), Mycobacterium avium complex (MAC), etc. · Mycobacterium leprae - the pathogen causing leprosy · Nontuberculous mycobacteria (NTM) - including all mycobacteria other than Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium leprae (M. leprae). · Mycobacterium tuberculosis group - the pathogen causing tuberculosis · Mycobacterium leprae group - the pathogen causing leprosy · Nontuberculous mycobacteria (NTM) - including all mycobacteria other than Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium leprae (M. leprae). Mycobacterium abscessus complex (MABSC), Mycobacterium avium complex (MAC), etc. · Mycobacterium abscessus complex (MABSC), Mycobacterium avium complex (MAC), etc. · Mycobacterium avium complex (MAC), etc.

[0013] Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis complex bacteria. TB is one of the oldest human infectious pathogens recorded in documents and still remains a major cause of increasing mortality and morbidity worldwide. The estimated number of new TB infections in 2015 was 10.4 million, and 1.4 million people died from active TB (see, for example, the World Health Organization (WHO) Global Tuberculosis Report 2016). In addition to such high morbidity and mortality, concerns about the emergence of multidrug-resistant tuberculosis (MDR-TB) are increasing, and in 2015, there were 580,000 cases. · Mycobacterium tuberculosis complex bacteria · one of the oldest human infectious pathogens recorded in documents and still remains a major cause of increasing mortality and morbidity worldwide · The estimated number of new TB infections in 2015 was 10 · 4 million, and 1.4 million people died from active TB (see, for example, the World Health Organization (WHO) Global Tuberculosis Report 2016). · World Health Organization (WHO) Global Tuberculosis Report 2016 · In addition to such high morbidity and mortality, concerns about the emergence of multidrug-resistant tuberculosis (MDR-TB) are increasing · and in 2015, there were 580,000 cases It has been shown that the patient was infected with drug-resistant TB. Coinfection with human immunodeficiency virus (HI V), etc. complicates treatment and was involved in 1.2 million TB cases in 2015 .

[0014] To treat multidrug-resistant bacteria (MDR) infections, the WHO has recommended implementing a 9- to 12-month treatment regimen using second-line anti-TB drugs. In such regimens as the 9- to 12-month Bangla desh regimen, combination therapy with gatifloxacin, ethambutol, pyrazinamide, and clofazimine is used for MDR-TB, and 87.9% of patients achieved relapse-free cure (for example, see Sotgiu, G, et al., “Applicability of the shorter ‘Bangladesh regimen’ in high multidr ug-resistant tuberculosis settings”, International Journal of Infectious Diseases (2017) 56 190-193). Sotgiu,G,et al.,“Applicability of the sh orter‘Bangladesh regimen’in high multidr ug-resistant tuberculosis settings”,Inte rnational Journal of Infectious Diseases (2017)56 190-193).

[0015] From other studies that investigated shortening the TB treatment period, it was shown that clofazimine has no clinical usefulness even after 2 weeks of oral administration (for example, Diacon, A.H., e t al., “Bactericidal Activity of Pyrazina mide and Clofazimine Alone and in Combin ations with Pretomanid and Bedaquiline”, ations with Pretomanid and Bedaquiline”, American Journal of Respiratory and Crit ical Care Medicine(2015),191(8),943-953 refer Based on the theory that this drug binds with high affinity to serum proteins in circulating blood, this lack of efficacy was considered to be due to low bioavailability of the drug. Although clofazimine has been empirically proven to be effective in the treatment of MDR-TB and extensively drug-resistant TB (XDR-TB), in short-term treatment, the bioactivity seems to be limited due to low bioavailability after systemic administration (for example, see Swanson, R.V., et al., “Pharmacokinetics and Pharmacodynamics of Clofazimine in a Mouse Model of Tuberculosis”, Antimicrobial Agents and Chemotherapy(2015), 59(6), 3042-3051).

[0016] Treatment of lung infections by inhaling antibiotics results in higher drug concentrations in the lungs and reduced side effects compared to systemic delivery (for example, see Touw, D.J., et al., “Inhalation of antibiotics in cystic fibrosis”, European Respiratory Journal(1995), 8, 1594-1604), and as a result, it is known that the bioactivity and efficacy are increased (for example, see Hickey, A.J., “Inhaled drug treatment for tuberculosis: Past progress and future prospects”, Journal of Controlled ​​​​​​​​​​​​​​​​See ed Release, (2016), 240, 127-134. TB infection model When clofazimine is aerosolized and administered to the model, compared with the case of oral administration of clofazimine, it has been demonstrated in an in vivo mouse model that the removal rate of bacilli is significantly improved only 28 days after the start of treatment (for example, see Verma , R.K., et al., “Inhaled microparticles con , R.K., et al., “Inhaled microparticles containing clofazimine are efficacious in t reatment of experimental Tuberculosis in Mice”, Antimicrobial Agents and Chemothe rapy (2013), 57(2), 1050-1052). The improvement in efficacy within this short period is presumably due to the fact that higher concentrations of clofazimine in pulmonary macrophages within tuberculous granulomas result from the direct delivery of clofazimine to the infection site in the lung. Therefore, aerosol administration of clofazimine to patients with MDR TB or XDR-TB infections should further improve the treatment outcome of the patients, and it may also shorten the period of the current treatment regimen.

[0017] Once, the group of non-tuberculous mycobacteria (NTM), which were formerly called atypical or ubiquitous mycobacteria, included more than 150 bacterial species. NTM are widely found in the natural environment and are rich in diversity. These are found not only in soil, the ground, and drinking water, but also may be detected in foods such as pasteurized milk and cheese at low temperatures. Usually, NTM are considered to have low pathogenicity.

[0018] and are rich in diversity. These are found not only in soil, the ground, and drinking water, but also may be detected in foods such as pasteurized milk and cheese at low temperatures. Usually, NTM are considered to have low pathogenicity. and are rich in diversity. These are found not only in soil, the ground, and drinking water, but also in foods such as pasteurized milk and cheese at low temperatures. Usually, NTM are considered to have low pathogenicity. are considered. However, these can be a cause of serious diseases in humans, especially those with weakened immunity or those already suffering from lung diseases. Currently, NTM are classified according to their growth rate and are divided into slow-growing (SGM) and rapidly growing (RGM) mycobacteria. The slow-growing Mycobacterium avium complex (MAC) includes the species Mycobacterium avium, Mycobacterium chimaera, and Mycobacterium intracellulare, which are the most important and most commonly seen pathogenic NTM. Likewise, most of these, such as Mycobacterium kansasii, Mycobaceterium malmoense, Mycobacterium xenopi, Mycobacterium simiae, Mycobacterium abscessus, Mycobacterium gordonae, Mycobacterium fortuitum, and Mycobacterium chelonae, also cause lung infections.

[0019] Mycobacterium marinum ​​​​​​​​​​​​​​Mycobacterium marinum is involved in skin and soft tissue infections such as aquarium granuloma.

[0020] In particular, RGM causes serious chronic lung diseases that are life-threatening, and are involved in disseminated, often fatal infections. Infections are usually caused by contaminated material involving catheters and invasive procedures, non-sterile surgical procedures or injections, as well as foreign body transplantation. Contact with showerheads and jacuzzis has also been reported to pose a risk of infection. NTM usually causes opportunistic infections in patients with chronic lung diseases such as chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF), and other patients with reduced immune function.

[0021] In recent years, the importance of Mycobacterium abscessus, a rapidly growing mycobacterium (RGM), as a human pathogen has become apparent. It includes the Mycobacterium abscessus complex (MABSC), which contains the subspecies Mycobacterium abscessus subsp. abscessus (M.a.abscessus (M·a·abscessus)), Mycobacterium abscessus bolletii, and Mycobacterium abscessus massiliense, and has a very high fatality rate compared to other RGMs.

[0022] ​​​​​​When CF patients are infected with Mycobacterium abscessus, lung destruction spreads, and in many cases, it is incurable, and the treatment failure rate is as high as 60 - 66%, which is particularly problematic. (For example, see Obregon-Henao A et al, Antimicrobial Agents and Chemotherapy, November 2015, Vol 59, No 11, p. 6904 - 6912; Qvist, T., Pressler, T., Hoiby, N. and Katzenstein, T.L., “Shifting paradigms of nontuberculous mycobacteria in cystic fibrosis”, Respiratory Research (2014), 15(1): pp. 41 - 47). When infected, lung destruction spreads, and in many cases, it is incurable, and the treatment failure rate is as high as 60 - 66%, which is particularly problematic. Since the treatment failure rate is as high as 60 - 66%, it is particularly problematic. (For example, see Obregon-Henao A et al, Antimicrobial Agents and Chemotherapy, November 2015, Vol 59, No 11, p. 6904 - 6912; Qvist, T., Pressler, T., Hoiby, N. and Katzenstein, T.L., “Shifting paradigms of nontuberculous mycobacteria in cystic fibrosis”, Respiratory Research (2014), 15(1): pp. 41 - 47). 6904 - 6912; Qvist, T., Pressler, T., Hoiby, N. and Katzenstein, T.L., “Shifting paradigms of nontuberculous mycobacteria in cystic fibrosis”, Respiratory Research (2014), 15(1): pp. 41 - 47). 6904 - 6912; Qvist, T., Pressler, T., Hoiby, N. and Katzenstein, T.L., “Shifting paradigms of nontuberculous mycobacteria in cystic fibrosis”, Respiratory Research (2014), 15(1): pp. 41 - 47). nd Katzenstein, T.L., “Shifting paradigms of nontuberculous mycobacteria in cystic fibrosis”, Respiratory Research (2014), 15(1): pp. 41 - 47). fibrosis”, Respiratory Research (2014), 15(1): pp. 41 - 47). fibrosis”, Respiratory Research (2014), 15(1): pp. 41 - 47). fibrosis”, Respiratory Research (2014), 15(1): pp. 41 - 47).

[0023] The association between NTM human infections and the AIDS pandemic is becoming even stronger. Mycobacteria belonging to the Mycobacterium avium complex (MAC) have been identified as the main causative agents of opportunistic infections in patients infected with the human immunodeficiency virus (HIV). The association between NTM human infections and the AIDS pandemic is becoming even stronger. Mycobacterium avium complex (MAC) have been identified as the main causative agents of opportunistic infections in patients infected with the human immunodeficiency virus (HIV). have been identified as the main causative agents of opportunistic infections in patients infected with the human immunodeficiency virus (HIV).

[0024] Some species of NTM are known to form biofilms. A biofilm is a small colony of bacteria encapsulated in an extracellular matrix, which confers stability and resistance to the human immune system. In recent years, some species of NTM have been shown to form biofilms that enhance their resistance to disinfectants and antibacterial agents. Biofilms is a small colony of bacteria encapsulated in an extracellular matrix, which confers stability and resistance to the human immune system. In recent years, some species of NTM have been shown to form biofilms that enhance their resistance to disinfectants and antibacterial agents. Biofilms is a small colony of bacteria encapsulated in an extracellular matrix, which confers stability and resistance to the human immune system. In recent years, some species of NTM have been shown to form biofilms that enhance their resistance to disinfectants and antibacterial agents. Biofilms is a small colony of bacteria encapsulated in an extracellular matrix, which confers stability and resistance to the human immune system. In recent years, some species of NTM have been shown to form biofilms that enhance their resistance to disinfectants and antibacterial agents. Biofilms The set progresses through several stages, including reversible adhesion, irreversible adhesion, biofilm formation by bacterial aggregation, organization, and signaling, and finally dispersion. As this process proceeds, bacteria construct a matrix containing extracellular polymeric substances (EPS) such as polysaccharides, lipids, and nucleic acids, forming a complex three-dimensional structure (see, for example, Sousa S. et al., International Journal of Mycobacteriology 4(2015), 36 - 43). Specifically, since mycobacteria do not produce extracellular polysaccharides, the EPS of mycobacteria differs in nature from other biofilms ( see, for example, Zambrano MM, Kolter R. Mycobacterial b iofilms: a greasy way to hold it together . Cell. 200). The biofilms of mycobacteria vary among species but may contain mycolic acid, glycopeptide lipids, mycolyl - diacylglycerol, lipooligosaccharide, lipopeptide tide, and extracellular DNA (Rose SJ, Babrak LM, Bermudez LE(2015) Mycobacterium avium Possesses Extracellular DNA that Contributes to Biofilm Formation, Structural Integrity, and Tolerance to Antibiotics. Review and original research from PLoS ONE). Aggregates within biofilms are known to enhance resistance to antibacterial agents ( see, for example, Faria S. et al., Journal of Pathogens, Vol 2015, Article ID 809014).

[0025] As a new approach to the treatment of NTM lung infection, aerosolized liposomal amikacin delivery / inhalation of amikacin solution sprayed by a jet nebulizer (Rose S.et al,2014,PLoS ONE,Volume 9,Issue 9,e 108703 and Olivier K.et al,Ann Am Thorac So c Vol 11,No 1,pp.30-35) In addition to dry powder microparticle inhalation for transpulmonary delivery of anti-TB drugs (Cholo M et al.,J Antimicrob Che mother.2012 Feb;67(2):290-8 and Fourie B . and Nettey O.,2015 Inhalation Magazine , Verma 2013 Antimicrob Agents Chemother ) has been proposed.

[0026] In the treatment of NTM lung disease, parenteral aminoglycosides, tigecycline, and line zolid, delamanid, and other promising oral antibacterial drugs such as bedaquiline, and in selected cases multidrug combination therapy regimens by inhaling amikacin following initial treatment by surgical intervention have been shown to be expected to produce results (Lu Ryu et al.,Tuberc Respir Dis 2016;79:74-84). However while the incidence and prevalence of NTM infections, particularly NTM lung disease, are increasing, and the treatment options are limited, it is necessary to develop new dosage forms / pharmaceutical preparations that improve the bioavailability of currently used antibacterial drugs such as clofazimine It is present. Inhalation can enhance efficacy and reduce side effects compared to oral and parenteral treatments. There is a possibility of doing so.

[0027] The combination of clofazimine and amikacin has been shown to act synergistically against both Mycobacterium abscessus (M ycobacterium abscessus) and Mycobacterium avium ( Mycobacterium avium) in vitro (for example, van Ingen, J., et al., “In Vitro Synergy between Clofazimine and A mikacin in Treatment of Nontuberculous M ycobacterial Disease”, Antimicrobial Agen ts and Chemotherapy 56(12), 6324 - 6327(201 2) see). Furthermore, the synergistic effect of combining clofazimine and bedaquiline against Mycobacterium tuberculosis (Mycobact erium tuberculosis) has also been shown (for example, Cokol, M.et al., “E fficient Measurement and factorization o f high - order drug interactions in Mycoba cterium tuberculosis”, Sciences Advances f high - order drug interactions in Mycoba cterium tuberculosis”, Sciences Advances 2017:3:e170881, 11 October 2017 see). Also, for Mycobacterium abscessus, a non - tuberculous acid - fast bacterium, the synergistic effect of the combination of clofazimine / bedaki lin has been shown (Ruth, M.M.et al., “A B edaquiline / Clofazimine Combination Regim en Might Add Activity to the Treatment o f Clinically Relevant Non-Tuberculous My cobacteria”,Journal of Antimicrobial Che motherapy(2019),doi.org / 10.1093 / jac / dky5 26).

[0028] Pathogenic fungi are becoming increasingly prominent as a major cause of human mortality. Current estimates suggest that deaths due to invasive fungal infections rival those of better-known infectious diseases such as tuberculosis. Candida albicans , Cryptococcus neoformans , and Aspergillus fumigatus are the most frequently encountered human pathogenic fungi. Although each of these species is involved in hundreds of thousands of infections annually, mortality rates are unacceptably high due to inadequate diagnostic methods and limited treatment options. Clofazimine has been shown to be effective against multiple fungal species as a combination agent (e.g., Robbins, N., et al., “An Antifungal Combination Matrix Identifies a Rich Pool of Adjuvant Molec ules that Enhance Drug Activity against Diverse Fungal Pathogens”,Cell Reports 1 al., “An Antifungal Combination Matrix Identifies a Rich Pool of Adjuvant Molec ules that Enhance Drug Activity against Diverse Fungal Pathogens”,Cell Reports 1 (see reference: 3,1481-1492, November 17, 2015). Fungi also act as commensal bacteria, colonizing bacteria, and / or pathogenic bacteria in cystic fibrosis (e.g., C hotirmall, S.H. and McElvaney, N.G., “Fungi in the cystic fibrosis lung: Bystanders o r pathogens?”, The International Journal of Biochemistry & Cell Biology 52(2014), 161-173, see reference).

SUMMARY OF THE INVENTION

PROBLEMS TO BE SOLVED BY THE INVENTION

[0029] Since crofazimine has low water solubility, its oral bioavailability is low and its microbial resistance is high. In order to formulate the drug in a liquid aqueous carrier so that aerosolization can be performed by a nebulizer for the purpose of depositing aerosol particles in the lower lung, special techniques for solubilizing and stabilizing the drug are also required.

MEANS FOR SOLVING THE PROBLEM

[0030] According to one embodiment of the present invention: (a) a therapeutically effective dose of crofazimine or a pharmaceutically acceptable derivative or salt thereof; (b) a nonionic surfactant having a hydrophilic-lipophilic balance value exceeding 10; (c) an aqueous liquid carrier selected from water, isotonic saline, buffered saline, and an aqueous electrolyte solution ; A pharmaceutical composition comprising ​​​​Clofazimine or a pharmaceutically acceptable derivative or salt thereof is in the form of particles in a suspension and is provided and the median diameter of the particles of clofazimine or a pharmaceutically acceptable derivative or salt thereof is less than 5 μm and the D90 is less than 6 μm, and a pharmaceutical composition is provided.

[0031] According to another embodiment of the present invention, the particles of clofazimine or a pharmaceutically acceptable derivative or salt thereof have an average diameter of less than 2 μm and a D90 of less than 3 μm.

[0032] In another embodiment of the present invention: (a) a therapeutically effective dose of clofazimine; (b) a nonionic surfactant having a hydrophilic-lipophilic balance value exceeding 10; (c) an aqueous liquid selected from water, isotonic saline, buffered saline, and an aqueous electrolyte solution as a carrier; and a pharmaceutical composition comprising clofazimine is provided in the form of particles in a suspension, and the particles of clofazimine have a median diameter of less than 5 μm and a D90 of less than 6 μm, and a pharmaceutical composition is provided.

[0033] In another embodiment of the present invention, the median diameter of the particles of clofazimine is less than 2 μm, and the D90 is less than 3 μm.

[0034] By aerosolizing the composition of the present invention with a suitable nebulizer, the delivery of aerosolized clofazimine to the lower respiratory tract (i.e., to the bronchi, bronchioles, and alveoli of the central and lower peripheral lungs) is significantly improved, thereby substantially improving the therapeutic effect. ral and lower peripheral lung) is significantly improved, thereby substantially improving the therapeutic effect.

[0035] More preferably, the inhalation device should be further adapted to locally deliver to the lungs an aerosol having an optimal particle size distribution for the purpose of uniformly depositing in the lower respiratory system. There should be.

[0036] Accordingly, the present invention provides an aerosol having a particle size that promotes delivery to the alveoli and bronchioles. A suitable aerodynamic particle size targeting the alveoli and bronchioles is between 1 and 5 μm. Larger particles than that selectively deposit in the upper respiratory system (upper lung), i.e., the bronchi and trachea and the mouth and pharynx, i.e., the oropharyngeal region. Accordingly, the inhalation device is adapted to generate an aerosol having an aerodynamic mass median diameter (MMAD) in the range of about 1 to about 5 μm, preferably in the range of about 1 to about 3 μm. In a further embodiment the particle size distribution is narrow and the geometric standard deviation (GSD) is less than about 2.5.

[0037] The present invention is based on the unexpected finding that by aerosol-administering clofazimine to the lungs in the form of a suspension, the active substance can be deposited in the lower (i.e., deep) respiratory system, thereby improving the bioavailability of a highly hydrophobic BCS class II active substance, and as a result significantly improving the therapeutic effect and reducing systemic side effects. Based on.

[0038] In another aspect, this finding improves antibacterial treatment for infections caused by mycobacteria and gram-positive bacteria, particularly pulmonary infections caused by NTM, such as CF, COPD, and opportunistic infections in patients with a compromised immune function (such as HIV patients), etc.

[0039] ​​​Furthermore, the present invention not only eliminates the systemic side effects of established oral treatment regimens for pulmonary infections caused by Gram-positive bacteria, particularly pulmonary TB and NTM infections, but also aims to reduce the dosage and treatment period of clofazimine.

[0040] It will be more appreciated by those skilled in the art that this application also discloses any combination of the individual features disclosed herein.

BRIEF DESCRIPTION OF THE INVENTION

[0041] DEFINITIONS The term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and properties of the compounds of the present invention and are not biologically or otherwise undesirable. In many cases, the compounds of the present invention can form salts of acids and / or bases by the presence of amino groups and / or carboxyl groups or groups similar thereto. Pharmaceutically acceptable acid addition salts can be formed using inorganic and organic acids. Examples of inorganic acids that can be used to derive salts include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Examples of organic acids that can be used to derive salts include, for example, acetic acid, propionic acid, naphthoic acid, oleic acid, palmitic acid, pamoic acid, stearic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, ascorbic acid, glucoheptonic acid, glucuronic acid, lactic acid, lactobionic acid, tartaric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0042] Pharmaceutically acceptable base addition salts can be formed using inorganic and organic bases. Examples of inorganic bases capable of inducing salts include sodium, potassium, lithium , ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc ; salts of ammonium, potassium, sodium, calcium, and magnesium are particularly preferred. Examples of organic bases capable of inducing salts include substituted amines such as primary, secondary , and tertiary amines, natural substituted amines, cyclic amines, basic ion exchange resins, etc . Specifically, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, histidine, arginine, lysine, benethamine , N-methyl-glucamine, and ethanolamine, etc. Other acids include dodecyl sulfuric acid , naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, and saccharin . According to the present invention, in addition to this free base, methanesulfonic acid, maleic acid, isonicotinic acid , nicotinic acid, malonic acid, and salicylate, particularly clofazimine methanesulfonate are preferably used.

[0043] According to the present invention, in addition to this free base, methanesulfonic acid, maleic acid, isonicotinic acid , nicotinic acid, malonic acid, and salicylate, particularly clofazimine methanesulfonate is preferably used.

[0044] As used herein, the term "pharmaceutically acceptable derivative" means, for example, the compounds disclosed in US Patent No. 9,540,336, and the entire disclosure of US Patent No. 9,5 40,336 is incorporated herein by reference as part of this specification . In addition to this, the meaning of the derivative is as described in Lu, Y., Zhen, M., Wang, B., Fu [[ID=�6]], L., Zhao, W., Li, P., Xu, J., Zhu, H., Jin, H., Yi , L., Zhao, W., Li, P., Xu, J., Zhu, H., Jin, H., Yi n, D., Huang, H., Upton, AM. and Ma, Z., "Clofaz Imine Analogs with Efficacy against Experimental Tuberculosis and Reduced Potential for Accumulation as described in “Antimicrobial Agents and Chemotherapy(2011),55(11):pp.5185- 5193. Furthermore, the “pharmaceutically acceptable derivatives” of the compounds are, for example, prodrugs of the above compounds. Generally, a prodrug is a derivative of the compound that can provide the active form of the compound after administration. This type of derivative can be, for example, an ester or amide of a carboxyl group, a carboxyl ester of a hydroxyl group, or a phosphate ester of a hydroxyl group. 5193. As further described in “Antimicrobial Agents and Chemotherapy(2011),55(11):pp.5185- 5193. Further, the “pharmaceutically acceptable derivatives” of the compounds are, for example, prodrugs of the above compounds. Generally, a prodrug is a derivative of the compound that can provide the active form of the compound after administration. This type of derivative can be, for example, an ester or amide of a carboxyl group, a carboxyl ester of a hydroxyl group, or a phosphate ester of a hydroxyl group. For example, prodrugs of the above compounds. Generally, a prodrug is a derivative of the compound that can provide the active form of the compound after administration. This type of derivative can be, for example, an ester or amide of a carboxyl group, a carboxyl ester of a hydroxyl group, or a phosphate ester of a hydroxyl group. For example, a derivative of the compound that can provide the active form of the compound after administration. This type of derivative can be, for example, an ester or amide of a carboxyl group, a carboxyl ester of a hydroxyl group, or a phosphate ester of a hydroxyl group. For example, it can be an ester or amide of a carboxyl group, a carboxyl ester of a hydroxyl group, or a phosphate ester of a hydroxyl group. Or a phosphate ester of a hydroxyl group.

[0045] “Therapeutically effective amount )”, “therapeutically effective dose e)”, or “pharmaceutically effective amount” means the amount at which clofazimine or its pharmaceutically acceptable salt or derivative disclosed in the present invention exhibits a therapeutic effect. The dose of clofazimine effective for treatment is a therapeutically effective amount. Therefore, the therapeutically effective amount used herein means the amount of clofazimine that results in the desired therapeutic effect, determined from clinical trial results and / or infection studies using animal models. As determined from clinical trial results and / or infection studies using animal models, it means the amount of clofazimine that results in the desired therapeutic effect.

[0046] The amount and daily dose of clofazimine can be mechanically determined by those skilled in the art, and several factors will vary, for example, depending on the specific microbial strains involved. Further, this amount may also depend on the patient's height, weight, sex, age, and medical history. A therapeutically effective amount in the case of prophylactic treatment is an amount effective to prevent microbial infection. The "therapeutic effect" is to relieve one or more symptoms of the infection to some extent, including curing the infection. "Curing" means that the symptoms of the active infection disappear,

[0047] including that the excessive portion of the viable bacteria involved in the infection completely or substantially disappears and becomes below the detection threshold by conventional measurements. However, certain long-term or permanent effects of the infectious disease may still exist even after a cure has been achieved (such as extensive tissue damage, etc.). As used herein, the "therapeutic effect" is defined as a statistically significant decrease in the amount of bacteria in the host, the expression of resistance, or the improvement of the infection symptoms determined by human clinical outcomes or animal experiments. The terms "treat", "treatment", or "treating" as used herein refer to administering a pharmaceutical composition / combination for prophylactic and / or therapeutic purposes. The term "prophylactic treatment" refers to treating patients who have not yet been infected but are susceptible to a specific infectious disease or are otherwise at risk. The term "therapeutic treatment" refers to treating patients who already have an infectious disease. Therefore, including the disappearance of the symptoms of the active infection, and including that the excessive portion of the viable bacteria involved in the infection completely or substantially disappears and becomes below the detection threshold by conventional measurements. However, certain long-term or permanent effects of the infectious disease may still exist even after a cure has been achieved (such as extensive tissue damage, etc.). As used herein, the "therapeutic effect" is defined as a statistically significant decrease in the amount of bacteria in the host, the expression of resistance, or the improvement of the infection symptoms determined by human clinical outcomes or animal experiments. The terms "treat", "treatment", or "treating" as used herein refer to administering a pharmaceutical composition / combination for prophylactic and / or therapeutic purposes. The term "prophylactic treatment" refers to treating patients who have not yet been infected but are susceptible to a specific infectious disease or are otherwise at risk. The term "therapeutic treatment" refers to treating patients who already have an infectious disease. Therefore, will vary, for example, depending on the specific microbial strains involved. Further, this amount may also depend on the patient's height, weight, sex, age, and medical history. A therapeutically effective amount in the case of prophylactic treatment is an amount effective to prevent microbial infection.

[0048] will vary, for example, depending on the specific microbial strains involved. Further, this amount may also depend on the patient's height, weight, sex, age, and medical history. A therapeutically effective amount in the case of prophylactic treatment is an amount effective to prevent microbial infection. The "therapeutic effect" is to relieve one or more symptoms of the infection to some extent, including curing the infection. "Curing" means that the symptoms of the active infection disappear,

[0049] The term "prophylactic treatment" refers to treating patients who have not yet been infected but are susceptible to a specific infectious disease or are otherwise at risk. The term "therapeutic treatment" refers to treating patients who already have an infectious disease. Therefore, will vary, for example, depending on the specific microbial strains involved. Further, this amount may also depend on the patient's height, weight, sex, age, and medical history. A therapeutically effective amount in the case of prophylactic treatment is an amount effective to prevent microbial infection. The "therapeutic effect" is to relieve one or more symptoms of the infection to some extent, including curing the infection. "Curing" means that the symptoms of the active infection disappear, In a preferred embodiment, the treatment is the administration of an effective amount of clofazimine to a mammal for treatment (either for therapeutic or prophylactic purposes).

[0050] Unless otherwise specified herein, the term "inhalation" means pulmonary inhalation.

[0051] Unless otherwise specified herein, the term "infection" as used herein means pulmonary infection.

[0052] Unless otherwise specified, the term "substantially" used to indicate the purity of a compound means that the purity of the compound exceeds 95%.

[0053] Unless otherwise specified, the term "appropriate particle size" refers to the particle size of clofazimine in a composition or the composition that provides the desired therapeutic effect when administered to a patient.

[0054] Unless otherwise specified, the term "appropriate concentration" refers to the concentration of a component in a composition or combination that provides a pharmaceutically acceptable composition or combination.

[0055] Pharmaceutical Compositions and Combination Medicines The following water quality grades are particularly suitable for the present invention: sterilized purified water, water for injection, sterile water for irrigation, sterile water for inhalation (USP), and those corresponding to water quality grades compliant with, for example, the European Pharmacopoeia or the National Formulary. , sterile water for inhalation (USP) and those corresponding to water quality grades compliant with, for example, the European Pharmacopoeia or the National Formulary ​​​​​​​​​​

[0056] According to the present invention, the aqueous electrolyte solution used as the aqueous liquid carrier further contains sodium chloride, potassium chloride, lithium chloride, magnesium chloride, calcium chloride, or a mixture thereof can be included.

[0057] The aqueous liquid carrier is preferably isotonic physiological saline (about 150 mM NaCl, preferably 0.9% NaCl corresponding to 154 mM NaCl).

[0058] It has been shown that clofazimine exists in at least four polymorphs (for example, see B annigan, et al., “Investigation into the S olid and Solution Properties of Known an d Novel Polymorphs of the Antimicrobial Molecule Clofazimine”, Cryst. Growth Des. 2 016, 16(12), pp. 7240 - 7250). Clofazimine can exist in the triclinic FI type, the monoclinic FII type, and the orthorhombic FIII type. Another type, FIV, has only been observed at high temperatures.

[0059] Therefore, in a further embodiment of the present invention: (a) a therapeutically effective dose of clofazimine; (b) a nonionic surfactant with a hydrophilic - lipophilic balance value exceeding 10; (c) an aqueous liquid carrier selected from water, isotonic physiological saline, buffered physiological saline, and an aqueous electrolyte solution; A pharmaceutical composition comprising clofazimine is provided in the form of particles in a suspension, and The median diameter of the particles of clofazimine is less than 5 μm, and D90 is less than 6 μm, preferably the median diameter is less than 2 μm, and D90 is less than 3 μm, and clofazimine is provided in one or more polymorphs selected from the triclinic FI type, the monoclinic FII type, and the orthorhombic FIII type, and mixtures of this kind of type A pharmaceutical composition is provided. In other embodiments, clofazimine is provided substantially in the orthorhombic FIII type.

[0060] In a further embodiment of the present invention, a pharmaceutical composition according to any of the composition embodiments described herein is provided, and the nonionic surfactant is polysorbate 20 (for example, T ween® 20), polysorbate 60 (for example, Tween® 60 ), polysorbate 80 (for example, Tween® 80), stearyl alcohol ), polyethylene glycol derivative of hydrogenated castor oil with a hydrophilic-lipophilic balance value of 14 to 16 (for example, Cremophor® RH40), polyethylene glycol derivative of hydrogenated castor oil with a hydrophilic-lipophilic balance value of 15 to 17 (for example, Cremophor® RH60), sorbitan monolaurate (for example, Span® 20), sorbitan monopalmitate (for example, Span® 40) ), sorbitan monostearate (for example, Span® 60), polyoxyethylene (20) oleyl ether (for example, Brij® 020), polyoxyethylene (20) cetyl ether (for example, Brij® 58), polyoxyethylene (10) cetyl ether (for example, Brij® C10), polyoxyethylene (20) cetyl ether (for example, Brij® 58), polyoxyethylene (10) cetyl ether (for example, Brij® C10), polyoxyethylene (20) cetyl ether (for example, Brij® 58), polyoxyethylene [[ID=__36]] (10) cetyl ether (for example, Brij® C10), polyoxyethylene Ren(10) oleyl ether (e.g., Brij® O10), polyoxyethylene (100) stearyl ether (e.g., Brij® S100), poly oxyethylene (10) stearyl ether (e.g., Brij® S10), polyoxyethylene (20) stearyl ether (e.g., Brij® S20 ), polyoxyethylene (4) lauryl ether (e.g., Brij® L4) , polyoxyethylene (20) cetyl ether (e.g., Brij® 93), polyoxyethylene (2) cetyl ether (e.g., Brij® S2), caproyl polyoxyl-8 glyceride (e.g., Labrasol®) , polyethylene glycol (20) stearate (e.g., Myrj™ 49), poly ethylene glycol (40) stearate (e.g., Myrj™ S40), poly ethylene glycol (100) stearate (e.g., Myrj™ S100), poly ethylene glycol (8) stearate (e.g., Myrj™ S8), and poly oxyl 40 stearate (e.g., Myrj™ 52), and mixtures thereof are selected.

[0061] In other embodiments of the present invention, a pharmaceutical composition according to any of the composition embodiments described herein is provided, the nonionic surfactant is polysorbate 80, and the aqueous liquid carrier is distilled water, hypertonic saline, or isotonic saline. In other embodiments of the present invention a pharmaceutical composition is provided, the hypertonic saline is 1% - 7% (w / v) sodium chloride . In a further embodiment of the present invention, a pharmaceutical composition is provided, the nonionic surfactant The surfactant is ultra-high purity polysorbate 80 (e.g., NOF Corporation Po lysorbate80 (Hx2)), and the aqueous liquid carrier is isotonic saline.

[0062] In another embodiment of the present invention, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, and the weight osmolality of the composition is in the range of 200 to 700 mOsm / kg In a further embodiment, the weight osmolality of the composition is in the range of 300 to 400 m Osm / kg. In a further embodiment of the present invention, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, and the nonionic surfactant is in the range of 0.001% to 5

[0063] %(v / v) of the total composition, and the amount of clofazimine is in the range of 0.1% to 20% (w / v) of the total composition. In another embodiment of the present invention, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, and the pharmaceutical composition comprises the following steps: / v) of the total composition.

[0064] In another embodiment of the present invention, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, and the pharmaceutical composition comprises the following steps: (1) Homogenizing a suspension of clofazimine, a nonionic surfactant, and water to obtain a suspension containing clofazimine with an appropriate particle size (1) Homogenizing a suspension of clofazimine, a nonionic surfactant, and water to obtain a suspension containing clofazimine with an appropriate particle size (2) Adjusting the pH of the suspension obtained from (1) to a pH between pH 5.5 and pH 7.5 (2) Adjusting the pH of the suspension obtained from (1) to a pH between pH 5.5 and pH 7.5 (3) Adjusting the sodium chloride concentration to an appropriate concentration (3) Adjusting the sodium chloride concentration to an appropriate concentration (4) Adjusting the weight osmolality to an appropriate level and is prepared by a process comprising.

[0065] In a further embodiment, the pH is adjusted to 7.4 and the sodium chloride concentration is 154 mM It is adjusted to sodium chloride. In other embodiments, the homogenization in step (1) is high pressure Homogenization, high shear homogenization, wet grinding, ultrasonic homogenization, or a combination of such treatments Is carried out. In other aspects, the homogenization of clofazimine is carried out by multi-stage homogenization In other embodiments, clofazimine with an appropriate particle size has an average diameter of less than 5 μm And particles with a D90 of less than 6 μm. In a further embodiment, clof Azimine is particles with an average diameter of less than 2 μm and a D90 of less than 3 μm.

[0066] In a further embodiment of the present invention, a pharmaceutical composition according to any one of the composition embodiments described herein Is provided, and the pharmaceutical composition comprises the following steps: (1) Homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension Containing clofazimine with an appropriate particle size; (2) Isolating clofazimine; (3) Adding clofazimine to a non-ionic surfactant and water; (4) Adjusting the pH of the suspension obtained from (3) to a pH between pH 5.5 and pH 7.5 Step; (5) Adjusting the sodium chloride concentration to an appropriate concentration; Is prepared by a process comprising.

[0067] In a further embodiment, the pH is adjusted to 7.4 and the sodium chloride concentration is adjusted to 154 mM sodium chloride In a further embodiment, the homogenization in step (1) is high pressure homogenization, high shear homogenization, wet grinding, ultrasonic homogenization, or a combination of such treatments Is carried out. In other embodiments, the homogenization of clofazimine is multi-stage homogenization Is carried out. In other embodiments, the homogenization of clofazimine is multi-stage homogenization It is implemented by pulverization. In other embodiments, clofazimine having an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In a further embodiment, clofazimine having an appropriate particle size has an average diameter of less than 2 μm and particles with a D90 of less than 3 μm. In a further embodiment, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, and this composition comprises the following steps: (1) pulverizing clofazimine to obtain clofazimine with an appropriate particle size; and

[0068] (2) adding clofazimine to a non-ionic surfactant and water; (3) adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5; (4) adjusting the sodium chloride concentration to an appropriate concentration. and (2) adding clofazimine to a non-ionic surfactant and water; (3) adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5; and (4) adjusting the sodium chloride concentration to an appropriate concentration. It is prepared by a process comprising.

[0069] In a further embodiment, the pH is adjusted to 7.4 and the sodium chloride concentration is adjusted to 154 mM sodium chloride. In other embodiments, the pulverization of clofazimine is carried out by jet milling, spray drying, ball milling, or supercritical fluid treatment. In other embodiments, the pulverization of clofazimine is carried out by multi-stage pulverization. In other embodiments, clofazimine with an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In a further embodiment, clofazimine with an appropriate particle size has an average diameter of less than 2 μm and particles with a D90 of less than 3 μm.

[0070] In other embodiments, the pulverization of clofazimine is carried out by jet milling, spray drying, ball milling, or supercritical fluid treatment. In other embodiments, the pulverization of clofazimine is carried out by multi-stage pulverization. In other embodiments, clofazimine with an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In a further embodiment, clofazimine with an appropriate particle size has an average diameter of less than 2 μm and particles with a D90 of less than 3 μm. In other embodiments, the pulverization of clofazimine is carried out by jet milling, spray drying, ball milling, or supercritical fluid treatment. In other embodiments, the pulverization of clofazimine is carried out by multi-stage pulverization. In other embodiments, clofazimine with an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In a further embodiment, clofazimine with an appropriate particle size has an average diameter of less than 2 μm and particles with a D90 of less than 3 μm. In other embodiments, the pulverization of clofazimine is carried out by multi-stage pulverization. In other embodiments, clofazimine with an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In a further embodiment, clofazimine with an appropriate particle size has an average diameter of less than 2 μm and particles with a D90 of less than 3 μm. In other embodiments, clofazimine with an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In a further embodiment, clofazimine with an appropriate particle size has an average diameter of less than 2 μm and particles with a D90 of less than 3 μm. In a further embodiment, clofazimine with an appropriate particle size has an average diameter of less than 2 μm and particles with a D90 of less than 3 μm.

[0071] In a further embodiment, according to any one of the composition embodiments described herein a pharmaceutical composition is provided, the composition comprising clofazimine, a non-ionic surfactant, an appropriate concentration of sodium chloride, and homogenizing a suspension in water adjusted to a pH between pH 5.5 and pH 7.5 to obtain a suspension of clofazimine having an appropriate particle size. The process is prepared by a process comprising In a further embodiment, the pH is adjusted to 7.4 and the sodium chloride concentration is adjusted to 154 mM sodium chloride. In a further embodiment, the homogenization is carried out by high-pressure homogenization, high-shear homogenization, wet milling, ultrasonic homogenization, or a combination of such treatments In other embodiments, the homogenization of clofazimine is carried out by multi-stage homogenization In other embodiments, clofazimine having an appropriate particle size has an average particle diameter of less than 5 μm and a D90 of less than 6 μm. In a further embodiment, an appropriate particle size of clofazimine consists of particles having an average diameter of less than 2 μm and a D90 of less than 3 μm there is.

[0072] In other embodiments, a process for preparing a pharmaceutical composition according to any of the composition embodiments described herein A process comprising the following steps: (1) Homogenizing a suspension of clofazimine, a non-ionic surfactant, and water to obtain a suspension containing clofazimine having an appropriate particle size step, and (2) Adjusting the pH of the suspension obtained from (1) to a pH between pH 5.5 and dpH 7.5 step, and (3) Adjusting the sodium chloride concentration to an appropriate concentration (4) Adjusting the weight osmolarity to an appropriate level A process is provided that includes.

[0073] In other embodiments, the pH is adjusted to 7.4 and the sodium chloride concentration is adjusted to 154 mM sodium chloride. In further embodiments, the homogenization is carried out by high-pressure homogenization, wet milling, ultrasonic homogenization, or a combination of such treatments. In further embodiments the homogenization of clofazimine is carried out by multi-stage homogenization. In further embodiments suitable particle size clofazimine has an average diameter of less than 5 μm and a D90 of less than 6 μm of particles. In other embodiments, suitable particle size clofazimine has an average diameter of 2 μ m and a D90 of less than 3 μm of particles.

[0074] In other embodiments, a process for preparing the embodiments of the pharmaceutical composition described herein, comprising: (1) homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension comprising suitable particle size clofazimine; (2) isolating the clofazimine; (3) adding the clofazimine to a non-ionic surfactant and water; (4) adjusting the pH of the suspension obtained from (3) to a pH between pH 5.5 and pH 7.5; (5) adjusting the sodium chloride concentration to a suitable concentration. (5) adjusting the sodium chloride concentration to a suitable concentration. A process is provided.

[0075] In other embodiments, the pH is adjusted to 7.4 and the sodium chloride concentration is adjusted to 154 mM sodium chloride. In further embodiments, the homogenization is carried out by high-pressure homogenization, wet milling, ultrasonic homogenization, or a combination of such treatments. In further embodiments , The homogenization of clofazimine is carried out by multi-stage homogenization. In a further embodiment wherein, the clofazimine with appropriate particle size has an average diameter of less than 5 μm and a D90 of less than 6 μm particles. In other embodiments, the clofazimine with appropriate particle size has an average diameter of 2 μ m and D90 of less than 3 μm particles.

[0076] In a further embodiment, a process for preparing a pharmaceutical composition according to any one of the pharmaceutical composition embodiments described herein, comprising the following steps: (1) micronizing clofazimine to obtain clofazimine with appropriate particle size step, and, (2) adding clofazimine to a nonionic surfactant and water (3) adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5 step, (4) adjusting the sodium chloride concentration to an appropriate concentration step,

[0077] In other embodiments, the pH is adjusted to 7.4 and the sodium chloride concentration is adjusted to 154 mM sodium chloride. In a further embodiment, the micronization of clofazimine is carried out by jet mill grinding, spray drying, ball mill grinding, or supercritical fluid treatment. In a further embodiment, the micronization of clofazimine is carried out by multi-stage micronization. In a further embodiment, the clofazimine with appropriate particle size has an average diameter of less than 5 μm and D90 of less than 6 μm particles. In other embodiments, the clofazimine with appropriate particle size has an average diameter of 2 μm and D90 of less than 3 μm particles.

[0078] In other embodiments, according to any one of the pharmaceutical composition embodiments described herein a process for preparing a pharmaceutical composition, comprising suspending clofazimine in water containing a nonionic surfactant and an appropriate concentration of sodium chloride, the pH of which is adjusted to between pH 5.5 and pH 7.5, and homogenizing the suspension to obtain clofazimine having an appropriate particle size is provided. In other embodiments, the pH is 7.4 and the appropriate concentration of sodium chloride is 154 mM sodium chloride. In a further embodiment, the homogenization is carried out by high-pressure homogenization, wet milling, ultrasonic homogenization, or a combination of such treatments is carried out. In a further embodiment, the homogenization of clofazimine is carried out by multi-stage homogenization. In a further embodiment, the clofazimine having an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In other embodiments, the clofazimine having an appropriate particle size has an average diameter of 2 μm and particles with a D90 of less than 3 μm. In a further embodiment, a process for preparing a pharmaceutical composition according to any one of the composition embodiments described herein, comprising the following steps: (a) homogenizing a suspension of clofazimine, a nonionic surfactant, and water to obtain a suspension containing clofazimine having an appropriate particle size; (b) adjusting the pH of the resulting suspension to a pH between pH 5.5 and pH 7.5; (c) adjusting the sodium chloride concentration to an appropriate concentration; and (d) adjusting the weight osmolality to an appropriate level, wherein steps (b), (c), and (d) may be carried out in the order of (b), (c), (d); (b), (d), (c is provided. In a further embodiment, the clofazimine having an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In other embodiments, the clofazimine having an appropriate particle size has an average diameter of 2 μm and particles with a D90 of less than 3 μm.

[0079] In a further embodiment, a process for preparing a pharmaceutical composition according to any one of the composition embodiments described herein, comprising the following steps: (a) homogenizing a suspension of clofazimine, a nonionic surfactant, and water to obtain a suspension containing clofazimine having an appropriate particle size; (b) adjusting the pH of the resulting suspension to a pH between pH 5.5 and pH 7.5; (c) adjusting the sodium chloride concentration to an appropriate concentration; and (d) adjusting the weight osmolality to an appropriate level, wherein steps (b), (c), and (d) may be carried out in the order of (b), (c), (d); (b), (d), (c is provided. In a further embodiment, the clofazimine having an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In other embodiments, the clofazimine having an appropriate particle size has an average diameter of 2 μm 7.5; (c) adjusting the sodium chloride concentration to an appropriate concentration; and (d) adjusting the weight osmolality to an appropriate level, wherein steps (b), (c), and (d) may be carried out in the order of (b), (c), (d); (b), (d), (c is provided. In a further embodiment, the clofazimine having an appropriate particle size has an average diameter of less than 5 μm and particles with a D90 of less than 6 μm. In other embodiments, the clofazimine having an appropriate particle size has an average diameter of 2 μm ); (c), (b), (d); (c), (d), (b); (d), (b), (c); or A process is provided that can be performed in the order of (d), (c), (b).

[0080] In other embodiments, a medicament according to any one of the composition embodiments described herein A process for preparing a composition, comprising the following steps: (a) Homogenizing clofazimine and a suspension in a non-aqueous liquid to obtain a suspension containing clofazimine with an appropriate particle size ; (b) isolating the clofazimine; (c) adding the clofazimine to a non-ionic surfactant and water; (d) adjusting the pH of the resulting suspension to a pH between pH 5.5 and pH 7.5; (e) adjusting the sodium chloride concentration to an appropriate concentration; and steps (d) and (e) can be performed in the order of (d ), (e); or (e), (d), and a process is provided.

[0081] In other embodiments, a process for preparing a pharmaceutical composition according to any one of the composition embodiments described herein comprises the following steps: (a) micronizing clofazimine to obtain clofazimine with an appropriate particle size, and (b) adding clofazimine to water containing a non-ionic surfactant, an appropriate concentration of sodium chloride, and having a pH adjusted to between pH 5.5 and 7.5, and a process is provided.

[0082] In other embodiments of the present invention, a combined medicament in the form of an inhalation aerosol comprising a composition according to any one of the composition embodiments described herein, using an ultrasonic nebulizer - I. A combined medicine is prepared by aerosolizing with a nebulizing device selected from an electrospray nebulizer, a vibrating membrane nebulizer, a jet nebulizer, and a mechanical soft mist metered inhaler. The aerosol particles generated by the nebulizing device have an aerodynamic mass median diameter of 1 - 5 μm. In a further embodiment, the inhaled aerosol is for deposition in the lower respiratory system. In other embodiments, the ejection rate of the nebulizing device is 0.1 - 1.0 ml / min. In other embodiments, the total inhalation volume is between 1 ml and 5 ml.

[0083] In other embodiments, it is a pharmaceutical composition according to any one of the composition embodiments described herein, comprising atomized 4 - 7% hypertonic saline, metaperiodate, sodium dodecyl sulfate, sodium bicarbonate, tromethamine, silver nanoparticles, bismuth - thiol, ethylenediaminetetraacetic acid, gentamicin loaded phosphatidylcholine - de corated gold nanoparticles, a chelating agent, cis - 2 - decenoic acid, D - amino acids, a peptide containing D - amino acid residues (D - enantiomeric peptide), gallium mesoporphyrin IX, gallium protoporphyrin I X, curcumin, patulin, penicillic acid, baicalein, naringenin, ursolic acid, asiatic acid, corosolic acid, fatty acids, host defense peptides, and antimicrobial peptides, an agent for dispersing and / or destroying biofilms, a mucolytic agent, and / or or in combination with a mucoactive agent and / or an agent that reduces biofilm formation. In another embodiment, a pharmaceutical composition for this use is provided. The substance is bedaquiline or its pharmaceutically acceptable salt or derivative, cefoxitin (ce foxitine), amikacin, clarithromycin, pyrazinamide, rifampin , moxifloxacin, levofloxacin, and para-aminosalicylates, and The compound is administered before, simultaneously with, or after the administration of an agent selected from the mixture of compounds listed above.

[0084] In other embodiments, a combination therapy according to any of the combination embodiments described herein. The drug is a nebulized mixture of 4-7% hypertonic saline, metaperiodate, sodium dodecyl sulfate, Thorium, sodium bicarbonate, tromethamine, silver nanoparticles, bismuth-thiol, ethyl Diaminetetraacetic acid, gentamicin-loaded phosphatidylcholine-modified gold nanoparticles, chiral cis-2-decenoic acid, D-amino acids, peptides containing D-amino acid residues, gallium Mesoporphyrin IX, Gallium Protoporphyrin IX, Curcumin, Patulin, Penicillin Syringic acid, baicalein, naringenin, ursolic acid, asiatic acid, corosolic acid, a biofilm-dissolving peptide selected from fatty acids, host defense peptides, and antimicrobial peptides; dispersing and / or disrupting agents, mucolytic and / or mucoactive agents, and / or A combination pharmaceutical for use in combination with an agent for reducing biofilm formation is provided. In another embodiment, the combination for this use comprises bedaquiline or a pharmaceutically acceptable salt thereof. acceptable salts or derivatives, cefoxitine, amikacin, Administering an agent selected from clarithromycin, pyrazinamide, rifampin, moxifloxacin, levofloxacin, and para-aminosalicylate, and mixtures thereof before, simultaneously with, or after, administering the composition of the present invention. In other embodiments the composition is administered before, simultaneously with, or after administering an agent selected from bedaquiline or a pharmaceutically acceptable salt or derivative thereof, and amikacin, and mixtures thereof In further embodiments, the composition is administered before, simultaneously with, or after administering bedaquiline or a pharmaceutically acceptable salt or derivative thereof In other embodiments, a pharmaceutical composition according to any one of the composition embodiments described herein is provided for use in the treatment and / or prevention of lung infections caused by mycobacteria or other gram-positive bacteria

[0085] In further embodiments, the infection is caused by a mycobacterium species selected from the group consisting of non-tuberculous mycobacteria and mycobacterium tuberculosis (Mycobacterium tuberculosis), and combinations thereof In further embodiments, the non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium leprae ​​​​​​​​e)) and selected from these combinations. In other embodiments, the infectious disease is pulmonary MAC disease and nontuberculous mycobacteriosis in patients with cystic fibrosis, chronic obstructive pulmonary disease, or acquired immunodeficiency syndrome selected opportunistic infection. In other embodiments, the infectious disease is an opportunistic infection by nontuberculous mycobacteria in patients with cystic fibrosis. In other embodiments, the composition for this use is administered before, simultaneously with, or after administration of an agent selected from bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazinamide, rifampin, moxifloxacin, levofloxacin, and para-aminosalicylate, and mixtures thereof. In other embodiments, the composition is administered before, simultaneously with, or after administration of an agent selected from bedaquiline or a pharmaceutically acceptable salt or derivative thereof, and amikacin, and mixtures thereof. In a further embodiment, the composition is administered before, simultaneously with, or after administration of bedaquiline or a pharmaceutically acceptable salt or derivative thereof. In other embodiments, a combination medicament according to any of the combination embodiments described herein is provided for use in the treatment and / or prevention of a pulmonary infection caused by mycobacteria or other gram-positive bacteria. In a further embodiment, the infectious disease is selected from nontuberculous mycobacteria and Mycobacterium tuberculosis and combinations thereof.

[0086] In other embodiments, a combination medicament according to any of the combination embodiments described herein is provided for use in the treatment and / or prevention of a pulmonary infection caused by mycobacteria or other gram-positive bacteria. In a further embodiment, the infectious disease is selected from nontuberculous mycobacteria and Mycobacterium tuberculosis and combinations thereof. In a further embodiment, the infectious disease is selected from nontuberculous mycobacteria and Mycobacterium tuberculosis (Mycobacterium tuberculosis) and combinations thereof. Caused by bacterial species belonging to the genus Mycobacterium In a further embodiment, the non-tuberculous mycobacteria are Mycobacterium avium , Mycobacterium intracellulare , Mycobacterium abscessus, and leprosy bacteria (Mycobacterium leprae), and combinations thereof. In other embodiments, the infection is a opportunistic infection selected from pulmonary MAC disease and non-tuberculous mycobacteriosis in patients with cystic fibrosis , chronic obstructive pulmonary disease, or acquired immunodeficiency syndrome. In other embodiments, the infection is an opportunistic infection caused by non-tuberculous mycobacteria in patients with cystic fibrosis . In other embodiments, the combination for this use is administered before, simultaneously, or after administering an agent selected from bedaquiline or a pharmaceutically acceptable salt or derivative thereof , cefoxitin, amikacin, clarithromycin, pyrazinamide , rifampin, moxifloxacin, levofloxacin, and para-aminosalicylic acid , and mixtures thereof, for use in administering the composition of the present invention. In other embodiments, the combination for this use is administered before, simultaneously, or after administering an agent selected from bedaquiline or a pharmaceutically acceptable salt or derivative thereof , amikacin, and mixtures thereof, for use in administering the composition of the present invention. In other embodiments, the combination for this use is administered before, simultaneously, or after administering bedaquiline or a pharmaceutically acceptable salt or derivative thereof , and the composition of the present invention . In other embodiments, the combination for this use is administered before, simultaneously, or after administering bedaquiline or a pharmaceutically acceptable salt or derivative thereof . In other embodiments, the combination for this use is administered before, simultaneously, or after administering bedaquiline or a pharmaceutically acceptable salt or derivative thereof, and the composition of the present invention . In other embodiments, the combination for this use is administered before, simultaneously, or after administering bedaquiline or a pharmaceutically acceptable salt or derivative thereof, and the composition of the present invention It is used for administering a composition.

[0087] In other embodiments, a system is provided for use in imparting antibacterial activity when treating or preventing pulmonary infections caused by Mycobacterium or other Gram-positive bacteria. This system comprises: 1) (a) a therapeutically effective amount of clofazimine; (b) a nonionic surfactant having a hydrophilic-lipophilic balance value exceeding 10; (c) an aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solutions and; a nebulized combined medicament comprising, 2) a nebulizer, wherein clofazimine is present in the form of a suspension, and the aerosol particles produced by this system have an aerodynamic mass median diameter of 1 - 5 μm.

[0088] In a further embodiment, a pharmaceutical composition according to any one of the composition embodiments described herein is provided for use in the treatment and / or prevention of pulmonary fungal infections or Clostridium difficile (clostridium difficile) or combinations thereof. In other embodiments, a pharmaceutical composition according to any one of the composition embodiments described herein is provided for use in the treatment and / or prevention of pulmonary fungal infections. In a further embodiment, the pulmonary fungal infection is Candida albicans (candi da albicans) or Aspergillus fumigatus (aspergilus fumigatus) or combinations thereof.

[0089] In a further embodiment, the pulmonary fungal infection or Clostridium difficile or A combination medicament according to any one of the embodiments described herein for use in the treatment and / or prevention of these combinations is provided. A combination medicament according to any one of the combination embodiments described herein for use in the treatment and / or prevention of pulmonary fungal infections is provided. In a further embodiment, the pulmonary fungal infection is candida albicans or aspergillus fumigatus, or a combination thereof. A combination medicament according to any one of the combination embodiments described herein for use in the treatment and / or prevention of pulmonary fungal infections is provided. In a further embodiment, the pulmonary fungal infection is candida albicans or aspergillus fumigatus, or a combination thereof. candida albicans) or aspergillus fumigatus (asper gilus fumigatus), or a combination thereof.

[0090] In another embodiment, a method of treating or preventing a pulmonary infection in a patient in need thereof, comprising administering by inhaling a composition according to any one of the composition embodiments described herein is provided. In another embodiment, the infection is caused by a mycobacterium species belonging to the genus mycobacterium, selected from the group of non-tuberculous mycobacteria and mycobacterium tuberculosis (Mycoba cterium tuberculosis)) and combinations thereof. In a further embodiment, the non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium leprae (Mycobacterium leprae) ), and combinations thereof. In a further embodiment, the infection is caused by a mycobacterium species belonging to the genus mycobacterium, selected from the group of non-tuberculous mycobacteria and mycobacterium tuberculosis (Mycoba cterium tuberculosis)) and combinations thereof. In a further embodiment, the non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium leprae (Mycobacterium leprae) ), and combinations thereof. In a further embodiment, the non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium leprae (Mycobacterium leprae) ), and combinations thereof. In a further embodiment, the infection is caused by a mycobacterium species belonging to the genus mycobacterium, selected from the group of non-tuberculous mycobacteria and mycobacterium tuberculosis (Mycoba ), and combinations thereof. In a further embodiment, the non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium leprae (Mycobacterium leprae) ), and combinations thereof. In a further embodiment, the infection is caused by a mycobacterium species belonging to the genus mycobacterium, selected from the group of non-tuberculous mycobacteria and mycobacterium tuberculosis (Mycoba Pulmonary MAC disease and pulmonary pulmonary disease in patients with cystic fibrosis, chronic obstructive pulmonary disease, or acquired immunodeficiency syndrome and nontuberculous mycobacterial infections. Mycobacterium tuberculosis is an opportunistic infection caused by nontuberculous mycobacteria in patients with cystic fibrosis.

[0091] In a further embodiment, the method comprises administering to a subject in need thereof a method for treating mycobacteria or other bacterial infections. A method for treating or preventing a pulmonary infection caused by gram-positive bacteria, comprising administering to a subject a pulmonary infection comprising administering to a subject a pulmonary infection caused by gram-positive bacteria, the method ... salts of phosphorus or its pharmaceutically acceptable derivatives, cefoxitine , amikacin, clarithromycin, pyrazinamide, rifampin, moxifloxacin levofloxacin, and para-aminosalicylates, and mixtures thereof. any of the composition embodiments described herein before, simultaneously with, or after administering the agent to be treated. In another embodiment, a method is provided, comprising administering by inhalation a composition according to one of the methods. In embodiments, the agent is bedaquiline or amikacin. , this agent is bedaquiline.

[0092] Particle size and distribution The therapeutic effect of aerosol therapy depends on the dose deposited and its distribution. The particle size of the aerosol determines the deposition amount and distribution of the drug aerosol in the lungs. It is one of the important variables.

[0093] In general, inhaled aerosol particles are dispersed by two mechanisms: collisions (usually with larger aerosols); deposition (dominated by smaller aerosol particles) and settling (dominated by smaller aerosol particles) Collisions occur when the momentum of inhaled aerosol particles is transferred to the airflow, causing the particles to settle. Occurs when large enough not to follow and reaches the physiological surface In contrast, sedimentation occurs when very small aerosol particles carried along with the inspiratory flow reach the physiological surface as a result of sedimentation mainly by the action of gravity in the lower respiratory system

[0094] Drug delivery to the lungs can be achieved by inhaling an aerosol through the oropharynx Aerosol particles with an aerodynamic diameter exceeding about 5 μm generally do not reach the lungs. Instead, they tend to collide deep in the throat and may be swallowed and absorbed orally Aerosol particles with a diameter of about 3 to about 5 μm are small enough to reach the upper-to-mid-pulmonary region (conducting airways), but are too large to reach the alveoli Smaller, i.e., about 0.5 to about 3 μm aerosol particles, can reach the alveolar region. Aerosol particles with a diameter smaller than about 0.5 μm tend to be exhaled with breathing during quiet breathing, but can also be deposited in the alveolar region by breath-holding

[0095] Aerosols used for pulmonary drug delivery are composed of a wide range of aerosol particle sizes, so statistical descriptors are used. Aerosols used for pulmonary drug delivery are usually characterized by their mass median diameter (MMD). That is, half of the mass contained in the aerosol particles is larger than the MMD, and half of the mass contained in the aerosol particles is smaller than the MMD. When the particle density is uniform, the volume median diameter (VMD) can be used interchangeably with the MMD. The determination of VMD and MMD is performed using laser diffraction. The width of the distribution is the geometric standard deviation ​​​​​​​​​​​​ It is represented by (GSD). On the other hand, the deposition of aerosol particles in the airway can be more accurately expressed using the aerodynamic diameter of the particles, so the aerodynamic mass median diameter is usually used. The MMA D measurement is performed by measuring inertial impaction or flight time. In the case of water particles, VMD, MMD , and MMAD should be equal. However, if the humidity is not controlled while the aerosol passes through the impactor, dehydration will occur, so the measured value of MMAD will be smaller than MMD and VM D. To use this description method, the measurements of VMD, MMD, and MM AD are considered to be performed under controlled conditions so that the descriptions of VMD, MMD, and MMAD are comparable (comparable ). However, for the purpose of explanation, the aerosol particle size of the aerosol particles shall comply with the United States Pharmacopeial Convention and be given as the MMAD determined by measuring at room temperature using a Next Generation Impactor (NGI) (In Pr

[0096] ocess Revision <601> Aerosols, Nasal Spra ys, Metered-Dose Inhalers, and Dry Powder Inhalers, Pharmacopeial Forum (2003), Volu me Number 29, pages 1176 - 1210, Jolyon Mitc hell, Mark Nagel “Particle Size Analysis of Aerosols from Medicinal Inhalers”, KON Inhalers, Pharmacopeial Forum (2003), Volume Number 29, pages 1176 - 1210, Jolyon Mitch ell, Mark Nagel “Particle Size Analysis of Aerosols from Medicinal Inhalers”, KON Inhalers, Pharmacopeial Forum (2003), Volume Number 29, pages 1176 - 1210, Jolyon Mitch A Powder and Particle Journal(2004), Volu me 22, pages 32 - 65 is also disclosed).

[0097] According to the present invention, the particle size of the aerosol is optimized so that clofazimine is maximally deposited at the infection site while maximizing tolerance. The aerosol particle size can be expressed in terms of the aerodynamic mass median diameter (MMAD). Large particles (e.g., MMAD > 5 μm ) are too large to travel along the bends of the airway and tend to deposit in the extrapulmonary and upper airway . When large particles deposit in the upper airway, intolerance symptoms (e.g., cough and bronchospasm) may appear .

[0098] Therefore, according to a preferred embodiment, the MMAD of the aerosol should be less than about 5 μm , preferably between about 1 - 5 μm, more preferably less than 3 μm (< 3 μm) .

[0099] However, by guiding the breathing method (guided breathing maneuver ), larger particles can be passed through the extrapulmonary and upper airway and penetrate deeper into the lungs than during quiet breathing, thereby increasing the deposition of the aerosol in the middle and lower respiratory systems . It is also possible to slow down the breathing method to 100 ml / min by guidance . Therefore, when using the guided breathing method, the preferred MMAD of the aerosol should be less than about 10 μm.

[0100] Another factor (other than the aerosol particle size) that is equally important is the particle size and size distribution of the solid particles , in this case the particle size and distribution of clofazimine. The solid particles of a given aerosol particle ​The particle size must be smaller than the aerosol particles containing it. Larger aerosol particles may contain one or more solid particles. Further, when dealing with dilute suspensions, most of the aerosol particles may not contain solid particles.

[0101] Therefore, it is desirable to use solid drug particles that are significantly smaller than the MMAD of the aerosol particles. For example, if the MMAD of the aerosol particles is 3 μm, it would be desirable for the solid particles to be below 1 μm.

[0102] Other considerations include, for example, the case of using a vibrating mesh nebulizer, where the formulation is delivered through the holes in the plate, and this plate breaks up the suspension into droplets. In that case, for such reasons, it is also necessary for the solid particles to be smaller than the holes so that they can pass

[0103] The particle size of the solid in the suspension can be given by the average diameter of the particles, or it can also be given by the particle distribution. The D90 value represents that 90% of the particles in the suspension are below its average diameter.

[0104] Nebulizers For aqueous and other non-pressurized liquid systems, various nebulizers (including small volume nebulizers) are available for aerosolizing the formulation. Compressor- driven nebulizers incorporate jet spray technology and use compressed air to generate a liquid aerosol. This type of device is, for example, , Healthdyne Technologies, Inc.; Invacare, I nc.; Mountain Medical Equipment, Inc.; Pari Respiratory, Inc.; Mada Medical, Inc.; Puri tan - Bennet; Schuco, Inc., DeVilbiss Health Care, Inc.; and are commercially available from Hospitak, Inc. Ultrasonic nebulizer Izers utilize mechanical energy by vibrating piezoelectric crystals to generate inhalable droplets, for example, and are commercially available from Omron Healthcare, Inc. and DeVil biss Health Care, Inc. Vibration mesh nebulizer Izers utilize either piezoelectric pulses or mechanical pulses to generate inhalable droplets. Other examples of nebulizers used in conjunction with clofazimine described herein are U.S. Patent No. 4,268,460; U.S. Patent No. 4,253,468; U.S. Patent No. 4,046,146; U.S. Patent No. 3,826,255; U.S. Patent No. 4,649,911; U.S. Patent No. 4,510,929; U.S. Patent No. 4 ,624,251; U.S. Patent No. 5,164,740; U.S. Patent No. 5,5 86,550; U.S. Patent No. 5,758,637; U.S. Patent No. 6,644 ,304; U.S. Patent No. 6,338,443; U.S. Patent No. 5,906,2 02; U.S. Patent No. 5,934,272; U.S. Patent No. 5,960,792 ; U.S. Patent No. 5,971,951; U.S. Patent No. 6,070,575; U.S. Patent No. 6,192,876; U.S. Patent No. 6,230,706 ; U.S. Patent No. 6,349,719; U.S. Patent No. 6,367,470; U.S. Patent No. 6,367,470; U.S. Patent No. 6,367,470; U.S. Patent No. 6,349,719; U.S. Patent No. 6,367,470; U.S. Japanese Patent No. 6,543,442; U.S. Patent No. 6,584,971; U.S. Patent No. 6,601,581; U.S. Patent No. 4,263,907; U.S. Patent No. 5,709,202; U.S. Patent No. 5,823,179; U.S. Patent No. 6, 192,876; U.S. Patent No. 6,644,304; U.S. Patent No. 5,54 9,102; U.S. Patent No. 6,083,922; U.S. Patent No. 6,161, 536; U.S. Patent No. 6,264,922; U.S. Patent No. 6,557,54 9; and U.S. Patent No. 6,612,303, the entire contents of all of which are hereby incorporated herein by reference as part of this specification. Examples of commercially available nebulizers that can be used with the clofazimine compositions described herein include Aero gen's Respirgard II®, Aeroneb®, Aeroneb® Pro, and Aeroneb® Go; Aradigm's AERx® and AERx Essence™; I-neb Resp ironics, Inc.'s Porta-Neb®, Freeway Fre edom™, Sidestream, Ventstream; and PARI, G mbH's PARI LCPlus®, PARI LC-Star® and e-Flow7m. Other non-limiting examples are disclosed in U.S. Patent No. 6,196,21 9.

[0105] According to the present invention, the pharmaceutical composition is preferably an ultrasonic nebulizer, an electrospray It can be aerosolized using an atomizing device selected from an isar, a vibrating membrane nebulizer, a jet nebulizer, or a mechanical soft mist inhaler. It can be aerosolized using an atomizing device selected from an isar, a vibrating membrane nebulizer, a jet nebulizer, or a mechanical soft mist inhaler.

[0106] Preferably, the device controls the patient's inhalation rate either electrically or mechanically. Preferably, the device controls the patient's inhalation rate either electrically or mechanically.

[0107] In a further preferred embodiment, the generation of the aerosol by the device is actuated by the patient's inhalation action, such as an AKITA device. In a further preferred embodiment, the generation of the aerosol by the device is actuated by the patient's inhalation action, such as an AKITA device.

[0108] Preferred (commercially available) examples of the above-mentioned nebulizer / devices used in accordance with the present invention are Vectura fox, Pari eFlow, Pari Trek S, Philips In nospire mini, Philips InnoSpire Go, Medspr ay device, Aeroneb Go, Aerogen Ultra, Respi ronics Aeroneb, Akita, Medspray Ecomyst, and Respimat. Respimat.

[0109] Use in treatment and / or prevention The pharmaceutical compositions and combination pharmaceuticals (aerosols, aerosolized formulations) and systems according to the present invention are for lungs caused by other crofazimine-sensitive bacteria such as Mycobacterium or Staphylococcus aureus (including methicillin-resistant and vancomycin-intermediate-resistant strains), Streptococcus pneumoniae, and Enterococcus spp. strains), Streptococcus pneumoniae, and Enterococcus spp. cus pneumoniae)), and Enterococcus spp. cus pneumoniae)), and Enterococcus spp. It is intended to be used for the treatment and / or prevention of infectious diseases. The pharmaceutical composition and combination medicaments of the present invention can also be used for the treatment and / or prevention of pulmonary fungal infections.

[0110] Administration of clofazimine According to the present invention, the pharmaceutical composition is delivered by atomizing about 1 to 5 ml, preferably 1 to 2 ml of the pharmaceutical composition of the present invention.

[0111] Therefore, based on the fact that the concentration of clofazimine in the pharmaceutical composition is about 20 mg / ml, the target fill dose is about 1 to 5 ml corresponding to 20 to 100 mg of clofazimine.

[0112] The daily lung dose (i.e., the amount of drug deposited in the lungs) of clofazimine administered according to the present invention is about 5 to 10 mg in the case of M. abscessus infection, which corresponds to a nominal dose of 15 to 30 mg (the amount of drug administered by the device).

[0113] Those skilled in the art will understand that the lung dose of clofazimine to be administered (and thus the fill dose / nominal dose / atomization volume) will be mechanically adjusted based on the minimum inhibitory concentration (MIC) of clofazimine against each specific strain, which is well established in the art.

[0114] Therefore, the daily lung dose will be divided according to the dosing frequency of once or twice a day.

[0115] According to the present invention, clofazimine is administered once or twice a day such that the total daily lung dose results in about 5 to 10 mg. ​​​​​​​

[0116] The amounts described above relate to the free base of clofazimine, and the dosages of derivatives and salts must be adjusted based on the MIC of each compound and strain, which will be apparent to those skilled in the art.

[0117] Mucolytic / biofilm modifying agent ) For the purpose of reducing the viscosity of sputum during aerosol therapy and destroying the existing biofilm, the treatment and / or prophylaxis according to the present invention can include the additional administration of a mucolytic and / or a biofilm disrupting agent. These agents can be formulated as a fixed combination, or administered simultaneously with or subsequent to a pharmaceutical composition / aerosol combination containing clofazimine according to the present invention.

[0118]

[0119] Agents for dispersing / destroying biofilms, mucolytics and / or mucus-active agents, and / or agents for reducing biofilm formation used according to the present invention are atomized 4 ~7% hypertonic saline, metaperiodate, sodium dodecyl sulfate, sodium bicarbonate, tromethamine, silver nanoparticles, bismuth-thiol, ethylenediaminetetraacetic acid, gentamicin loaded phosphatidylcholine-modified gold nanoparticles, chelating agents, cis-2-decenoic acid, D-amino acids, peptides containing D-amino acid residues, gallium mesoporphyrin IX, gallium protoporphyrin IX, curcumin, patulin, penicillic acid, baicalein, naringenin, ursolic acid, asiatic acid, corosolic acid, fatty acids, host defense peptides, and antimicrobial peptides.

[0120] Additionally, other pharmaceutically active agents may be used in combination with the pharmaceutical composition / aerosol combination according to the present invention. Such active agents include bedaquiline or its pharmaceutically acceptable salts or derivatives. Cefoxitine, amikacin, clarithromycin, pyramidalisol Zinamide, rifampin, moxifloxacin, levofloxacin, and para-amino salicylates, as well as mixtures thereof.

[0121] These agents can be formulated as a combination or can be used in combination with the clopidogrel of the present invention. It may also be administered before, simultaneously with, or after the pharmaceutical composition / aerosol combination containing Fazimin. Cut.

[0122] The following examples more fully illustrate the manner of using the above-described invention and further illustrate the principles of the invention. It serves to illustrate the best modes contemplated for carrying out various aspects of the invention. Embodiments in accordance with the present invention are encompassed by the claims set forth herein. [Example]

[0123] test The following exemplary compositions and combinations were prepared according to the processes described herein.

[0124] Example 1 Clofazimine (as triclinic form I) 200 mg, sodium chloride 90 mg, and water 9.5 ml was homogenized in an Ultra-Turrax homogenizer at 10,000 rpm for 5 minutes. Mixing was performed twice. 0.5 ml of polysorbate 80 (NOF Hx2) was added. The mixture was subjected to an ultrasonic probe (Bandelin S Branson Digital with onoplus Probe MS73 was treated 7 times (3 minutes each) at 70% amplitude with a Sonifier (trademark) 250D. The volume was adjusted to 10 ml with water. This suspension was filtered through a VWR folded qualitative filter paper (303, particle retention capacity 5 - 13 μm, size: 150 mm) to obtain the composition of Example 1. The median particle size of crophazimine in the composition of Example 1 was 3.9 μm, and the D90 was 6.7 μm was. The concentration of crophazimine was measured at 280 nm by ultraviolet / visible spectrophotometry and calibrated with a 1 mg / ml stock solution of crophazimine diluted in the mobile phase, and was determined to be 7.16 mg / ml.

[0125] The composition of Example 1 is shown in Table 1.

[0126]

Table 1

[0127] Preparation of orthorhombic form III crophazimine A slurry of crophazimine (10 g) in toluene (20 ml) was stirred at 800 rpm for 72 hours in an oil bath at 40 °C using a magnetic stirrer. The solid part of the slurry was filtered and recovered with a crucible filter, and vacuum dried in an oven with the maximum temperature set at 40 °C and. This yielded 8.64 g of substantially pure (≥98%) orthorhombic form III croph azimine. was obtained.

[0128] Example 2 A suspension containing 6 g of orthorhombic form III crophazimine in 100 ml of water containing 0.5% polysorbate 80 (NOF Hx2) and 0.6% sodium chloride was prepared with an Ultra - T ​Pre - micronized at about 10,000 rpm for about 40 seconds using urrax (registered trademark) . A preliminary preparation was carried out by adding water containing 0.6% sodium chloride to make the volume 300 ml . 300 ml of this suspension was fed into the inlet of a homogenizer, M - 110EH - 30 m icrofluidizer (Microfluidics, Westwood, MA, USA), and the suspension was circulated through the H30Z chamber at 5,000 psi for 15 minutes to perform a preliminary homogenization step. Then, a second H10Z chamber was attached in series to the first chamber, and the suspension was further homogenized at 25,000 psi for 23 minutes . When particle size analysis was performed using HORIBA LA 950, it was shown that the median particle size was 0 .83 μm and the D90 value was 1.17 μm. Measured at 280 nm by ultraviolet / visible spectroscopy and calibrated with a 1 mg / ml stock solution of crofazimine diluted with the mobile phase , the concentration of crofazimine was determined to be 16.05 mg / ml .

[0129] The composition of Example 2 is shown in Table 2

[0130]

Table 2

[0131] Example 3 A suspension in which crofazimine (crystal modification: orthorhombic system type III) was suspended in a solution of water, sodium chloride, and polysorbate 80 was used with an M - 110EH - 30 Microflui dizer (registered trademark) Processor (chambers: H30Z and G10Z), arranged in an H30Z - G10Z configuration, and operated at a pressure of 28,250 psi for 30 minutes . . The composition of Example 3 was produced. The median particle size of the obtained clofazimine particles was 1.28 μ m, and the D90 was less than 2 μm.

[0132] The composition of Example 3 is shown in Table 3.

[0133]

Table 3

[0134] Viscosity measurement The viscosity of the composition of Example 3 was measured in stress control mode using a STRESSTECH Rheometer. A double gap geometry was utilized and the spindle was rotated continuously to ensure that the fine particles remained suspended at each temperature point (during temperature points). The viscosity measurement was carried out with stresses of 0 .01, 0.05, and 0.1 Pa, and measured at 20 °C, 25 °C, and 30 °C respectively The average value of the viscosity obtained by performing the measurement with two separate sample loadings is shown in Table 4 below.

Table 4

[0135]

Table 4

[0136] Animal model and efficacy test Rather than administering clofazimine systemically, preclinical data was obtained to establish the concentration levels in lung tissue after direct delivery to the respiratory tract. For this purpose, the ability of the composition of the present invention to inhibit the growth of clinical (clinical al) NTM bacterial species was tested in an in vivo acute pulmonary infection mouse model of NTM. To investigate NTM lung infections for each target bacterial species, two species of Separate mouse models were used. To conduct the test, Mycobacterium avium ( M. ycobacterium avium,) strain 2285 and Mycobacterium abscessus strain 103 were used ( For details of the strains, refer to "Phylogenetic analysis of Mycobac terial species using whole genome sequen ces" registered in the EMBL / GenBank / DDBJ database (September 2014). Hazbon M.H., Riojas M.A., Damon A.M., Alalade R., Cantwell B.J., Monaco A., King S., Sohrabi A.). These two mycobacterial species have already been used as models of NTM infection in the literature (Obregon-Henao et al. 2015 Antimicrob Agents Chemother; and C han et al. Animal Models of Non-Tuberculo us Mycobacterial Infections, Mycobact Dis 2016).

[0137] In vivo safety test in Balb / C mice: To evaluate in vivo safety and tolerance, 6- to 8-week-old Balb / C female mice were obtained from Charles River. The mice were allowed to rest for one week before dosing. Three healthy mice were provided for each dose of clofazimine and administered a total of three times every other Performed. The aerosol of this compound was administered intratracheally to three healthy mice three times at one-day intervals using a Microsprayer (registered trademark).

[0138] The safety of clofazimine at 20 mg / kg (forced oral administration, 200 μl) has been confirmed . The composition of Example 1 showed no toxicity even at the maximum dose tested (10.0 mg / kg; 0.2506 mg / dose, in 35 μl, intratracheally). Therefore, the composition of Formula I is considered safe at 1 0.0 mg / kg and shows sufficient tolerance.

[0139] Determination of minimum growth inhibitory concentration The minimum inhibitory concentration (MIC) test was performed by the microbroth dilution method using Mueller-Hinton (MH) broth (cation adjusted )(calcium and magnesium ion concentrations recommended by CLSI standard M7-A7 )(Becton Dickinson). The MIC test was also performed by the microbroth dilution method using 7H9 broth (Sigma-Aldrich). The rationale for using both MH and 7H9 broths for compound screening is that antimycobacterial compounds show different MIC activities depending on the broth used in the MIC test, as has been found .

[0140] M. abscessus was grown on 7H11 agar plates (Sigma-Al drich) in ambient air at 35 - 37 °C (varying depending on the bacterial strain) for 3 days, and M. avium was grown on 7H11 agar plates (Sigma-Aldrich) in ambient air at 37 °C for 21 - 30 days. .

[0141] Colony forming units (CFU) were collected from agar plates and 0.05% Tween-80 was added. On day 3, the cells were added to either MH broth or 7H9 broth supplemented with M. abscessus (M Optical microscope (M. abscessus) or after the 12th day (M. avium) The density (OD) was measured by absorbance and kept between 0.08 and 0.1 (McFarland turbidity standard). The bacteria were grown in air at 35-37°C until they reached a concentration of 0.5. The suspension is adjusted to an OD of 0.08-0.1 (McFarland turbidity standard solution No. 0.5). The compound was suspended in DMSO at a concentration of 1.28 mg / ml. Compound stock solutions were prepared by turbidity, with the test range set at 64 to 0.062 μg / ml. Then, 180 μl of broth (either MH or 7H9) was added to a 96-well plate. In addition to the first row of the plate, 100 μl of broth was added to the remaining rows of the 96-well plate. 20 μl of the compound stock solution was added to the first row of wells and serially diluted. 100 μl of the solution specific for each microorganism was added to all wells except for the medium-only control wells. QC Reagents: 1) Bacteria-only negative control; 2) Media-only negative control; 3) Clarithromycin positive control. Sex-drug control.

[0142] The OD of M. abscessus was measured after 3 days, and M. avium (M. avium) was measured after 12 days. Following these measurements, Resaz Plates were measured using the Urin Microtiter Assay Plate method. Briefly, this method involves the extraction of resazurin (7-hydroxy-3H-phenoxazine-3- It is to add (10-oxide) to a 96-well plate. Resazurin is a blue pigment, which itself emits weak fluorescence, but when irreversibly reduced, it becomes resorufin that emits strong red fluorescence with a pink color. This is used as a redox indicator for determining the viability of bacterial cells in MIC measurement. It is a blue pigment, which itself emits weak fluorescence, but when irreversibly reduced, it becomes resorufin that emits strong red fluorescence with a pink color. This is used as a redox indicator for determining the viability of bacterial cells in MIC measurement.

[0143] The tests were carried out in triplicate. Test #1 was carried out after storing the composition of Example 1 at 4°C for 2 months, Test #2 was carried out after 4 months, and Test #3 was carried out after 5 months.

[0144] Minimum inhibitory concentration in the presence and absence of CF sputum The minimum inhibitory concentration measurement was carried out as described above.

[0145] To investigate the effect of sputum from patients with cystic fibrosis (CF) on the antibacterial activity of clofazimine (CFZ) and the composition of Example 1, sputum was collected from patients who had not received antibacterial drugs within 48 hours, and the sputum was sterilized by exposure to UV light to remove endogenous bacteria. After sterilization, M. abscessus, M. avium (M. avium), M. intracellulare, and M. Chimaera were incubated in 10% CF sputum and then the MIC test was performed. According to the same CLSI protocol as described above, the MIC of the composition of Example 1 was measured in the presence and absence of sputum from patients with cystic fibrosis. All the tests were carried out in duplicate.

[0146] The MIC values of clofazimine and the composition of Example 1 in the presence and absence of sputum are shown in Table 5.

[0147] ​​​​​​

Table 5

[0148] From the results shown in Table 5, it can be seen that both clofazimine and the composition of Example 1 exhibit stable MICs against a wide range of non-tuberculous acid-fast bacterial species.

[0149] These data suggest that the composition of Example 1 exhibits potent in vitro activity against both M. abscessus (M.abscessus) and M. avium, and is stable for at least this period.

[0150] Mouse model of M. abscessus in SCID mice Six- to eight-week-old SCID female mice were purchased from Charles River. The mice were allowed to rest for one week before infection.

[0151] A 1-ml aliquot of the working stock of M. abscessus 103 strain was taken and stored at -80 °C until use. For infection, the aliquot was thawed and diluted with sterile 1×PBS using a 1-ml Luer-lock syringe fitted with a 26-gauge needle and disrupted 20 times.

[0152] Acute-infected SCID mouse models were infected with 1×10 6 CFU / mouse (M. abscessus 103 strain) by non-invasive intratracheal injection.

[0153] Three mice were sacrificed on day 1 after infection to determine the bacterial uptake. The whole lungs, spleens, and livers were excised and homogenized in 4.5 ml of 1×PBS Homogenized. The homogenate was serially diluted at a dilution ratio of 1:10, and the diluted solutions (0-1-2- 3-4-5-6-7) were spread on 7H11 agar plates. The plates were left standing in a dry air incubator at 32°C (varying depending on the strain) for 7 days.

[0154] 10.0 mg / kg of the composition of Example 1 was administered via the transpulmonary route using a Microsprayer (registered trademark) (35 μl), and clofazimine (forced oral administration), amikacin (subcutaneous ) were administered in an amount of 200 μl per mouse. These were started on the 2nd day after infection and continued every other day for 8 days.

[0155] The mice were sacrificed 2 days after the last dose of the compound was administered. Six mice from all groups (untreated control, clofazimine (forced oral administration), the composition of Example 1, and amikacin -treated mice) were sacrificed, and the bacterial load was measured. Lung homogenates were plated at 0-1-2-3-4-5-6 -7, spleens at 0-1-2-3-4-5-6-7, and livers at 0-1-2-3-4-5-6 -7.

[0156] A protection value expressed as Log10 of at least 0. 60 suggests that the activity is statistically significant. Statistical analysis was performed by first logarithmically transforming the CFU and then evaluating this by one-way ANOVA, followed by one- way Tukey test for analysis of variance by multiple comparison (GraphPad Prism analysis software). Differences were considered significant at a 95% confidence level.

[0157] Table 6 shows the results of infecting SCID mice with M. abscessus ​​​​​​Subsequent Log 10 Shows the mean value of CFU data and the standard error of the mean (SEM). "n" is The total number of animals per group at the time of slaughter.

[0158]

Table 6

[0159] The data in Table 6 show that the amount of bacteria recovered from the lungs and spleens of animals infected with M. abscessus was significantly reduced by treatment with the composition of Example 1. This reduction in the amount of bacteria was statistically improved compared to treatment with amikacin or oral clofazimine.

[0160] Mouse model of Beige mice infected with M. avium Six to eight-week-old female Beige mice were purchased from Charles River. Before infection, the mice were allowed to rest for one week.

[0161] In the acute infection Beige mouse model, 1×10 8 colony-forming units (CFU) / ml (M · avium (M. avium) strain 2285 rough type) was non-invasively lung-infected by exposure to aerosol. The test stock solution of M. avium (M. avium) strain 2285 rough type was aliquoted 1 ml and frozen, and stored at -80 °C until use. For infection, the aliquot was thawed, and a 1 ml Luer-lock syringe with a 26 g needle was used to disrupt it 20 times and diluted with sterile 1× phosphate-buffered saline.

[0162] On the 1st and 7th days after infection, three mice were sacrificed and bacterial uptake was measured. Whole lungs, The spleen and liver were excised, homogenized in 4.5 ml of 1×PBS, and diluted 1:10. The dilutions (0-1-2-3-4-5-6-7) were plated on 7H11 / OADC, TSA, and Middlebrook agar plates and incubated for 30 days in a dry air incubator at 32 °C (varies according to the strain).

[0163] 10.0 mg / kg of the composition of Example 1 was administered via the transpulmonary route (35 μl) using a Microsprayer®, and clofazimine (forced oral administration) was administered to each mouse in an amount of 200 μl. These were started on day 7 after infection and continued every other day for 10 days.

[0164] The mice were sacrificed 5 days after administration of the final dose of the compound. Six mice from all groups (untreated control, clofazimine (forced oral administration), and the composition of Example 1) were sacrificed and the bacterial load was measured. 0-1-2-3-4-5-6-7 of the lung homogenate, 0-1-2-3-4-5-6-7 of the spleen, and 0-1-2-3-4-5-6-7 of the liver were plated on plates.

[0165] A defensive value expressed as Log10 of at least 0.60 suggests that the activity is statistically significant. Statistical analysis was performed by first transforming the CFU to logarithms and then evaluating this by one-way ANOVA followed by multiple comparison by one-way Tukey test (using the SigmaStat software program). Differences were considered significant at a 95% confidence level.

[0166] 10Shows the average value of CFU data.

[0167]

Table 7

[0168] The data in Table 7 show that the amount of bacteria recovered from the lungs and spleens of animals infected with M. avium m was significantly reduced by treatment with the composition of Example 1. .

[0169] Chronic infection Beige mouse model Six- to eight-week-old Beige mice were rested for one week before infection. On day 0, M. avium M. avium strain 2285 rough type at 1 × 10 8 CFU was used to infect the mice by lung infection. On day 1, three mice were sacrificed, and on day 27, six mice were sacrificed to measure bacterial uptake and the amount of bacteria before treatment. The whole lungs, spleens, and livers were excised, homogenized in 4.5 ml of 1× PBS, and diluted (0-1-2-3-4-5-6-7) was plated on 7H11 agar plates and charcoal agar plates. The plates were left standing in a 37°C dry air incubator for 25 to 30 days.

[0170] The remaining infected Beige mice were treated every other day starting on day 28 for a total of 14 times. The animals were treated with one of the following treatments: physiological saline (Microsp rayer (registered trademark), 35 μl); clofazimine (forced oral administration, 20 mg / kg , 200 μl); the composition of Example 1 (IT, Microsprayer (registered trademark), 1 0 mg / kg, 35 μl).

[0171] The mice were sacrificed on day 57, two days after the last treatment. The plates were placed in a 37°C dry air It was left standing in the incubator for 30 days.

[0172] For statistical analysis, first the CFU was logarithmically transformed, and then this was evaluated by one-way ANOVA After evaluation by one-way ANOVA, analysis of variance by multiple comparison was performed by one-way Tukey test This was done by doing so. A difference is considered significant at a 95% confidence level.

[0173] Table 8 shows the mean values of the Log og 10 CFU data after chronic infection of Beige mice with M. avium.

[0174]

Table 8

[0175] These data suggest that clofazimine hardly penetrates into the granuloma-like structures formed in the established "chronic" NTM-infected animal model. The composition of the present invention does not have this problem and seems to be able to maintain antibacterial activity even after the infection has been fully established.

[0176] The integrity of the barrier function and the effect on inflammation after exposing the composition of Example 3 to lung epithelial cells in vitro Cell viability To evaluate the cell viability of lung epithelial cells, three different types of cells: Calu-3; A 549; and hAELVi cells were used under two in vitro conditions. The cells were either in the "immersion condition" (i.e., in the cell culture medium on a Transwell (trademark) plate) or in the "air-liquid interface" (ALI) simulation condition where the cell culture medium was removed from the apical side of the cells. In the "immersion condition," Calu-3 cells were immersed in the composition of Example 3 at three doses (10%, 5%). The cells were exposed to 0% or 100% HCl for 4 hours. Staining was performed using fluorescein orange / propidium iodide (AO / PI) staining to identify live / dead cells. Red fluorescence indicates dead cells.

[0177] Uptake by macrophages THP-1 cells were incubated with 124 ng of phorbol 12-myristate 13-acetate (PMA). The cells were incubated with 100 μg / ml of erythrocyte monolayer for 3 days to differentiate into macrophage-like cells. Once ripe, add the composition of Example 3 (diluted 1:200 with Hank's Balanced Salt Solution (HBSS)) for 4 minutes. The cells were stained with AO / PI as described above to confirm the cells after exposure. Survival rates were determined.

[0178] Transepithelial Electrical Resistance (TEER) measurement 1 × 10 Calu-3 cells 5 Transwell (commercial) The cells were seeded onto a 3460 plate and grown for 12 days until they reached confluence. World Precision Instruments, Friedberg, Ge TEER measurements were performed using a Calu (Calmany) according to the manufacturer's instructions. -3 cells were incubated with saline (negative control) or the composition of Example 3 (concentration: 20 mg / ml, 10 After 2-4 hours of exposure, the cells were exposed to either 100mg / ml or 2mg / ml of erythrocytes. The TEER was measured.

[0179] Inflammatory cytokine production Differentiated THP-1 cells (dTHP-1) were exposed to the composition of Example 3 for 4 hours or 24 hours. Exposed (diluted 1:200 in HBSS). Exposure to HBSS alone was used as a negative control and lipopolysaccharide (LPS) (100 ng / ml) was administered as a positive control.

[0180] After incubation for t = 4 hours or 24 hours, the supernatant was removed from the cells and pooled. Enzyme-linked immunosorbent assay (ELISA) was performed on the pooled supernatant samples. Separate ELISA kits were used for TNF-α, IL-6, IL-8, and IL-10 according to the manufacturer's instructions. After incubation for t = 4 hours or 24 hours, the supernatant was removed from the cells and pooled. Enzyme-linked immunosorbent assay (ELISA) was performed on the pooled supernatant samples. Separate ELISA kits were used for TNF-α, IL-6, IL-8, and IL-10 according to the manufacturer's instructions. After incubation for t = 4 hours or 24 hours, the supernatant was removed from the cells and pooled. Enzyme-linked immunosorbent assay (ELISA) was performed on the pooled supernatant samples. Separate ELISA kits were used for TNF-α, IL-6, IL-8, and IL-10 according to the manufacturer's instructions. After incubation for t = 4 hours or 24 hours, the supernatant was removed from the cells and pooled. Enzyme-linked immunosorbent assay (ELISA) was performed on the pooled supernatant samples. Separate ELISA kits were used for TNF-α, IL-6, IL-8, and IL-10 according to the manufacturer's instructions.

[0181] After performing one-way analysis of variance (ANOVA), statistical analysis was performed using Tukey post-hoc test. A p-value < 0.05 was considered statistically significant. After performing one-way analysis of variance (ANOVA), statistical analysis was performed using Tukey post-hoc test. A p-value < 0.05 was considered statistically significant. After performing one-way analysis of variance (ANOVA), statistical analysis was performed using Tukey post-hoc test. A p-value < 0.05 was considered statistically significant.

[0182] Results Even after incubation for 4 hours under the "immersion" condition, the composition of Example 3 did not visually reduce cell viability at any of the administered concentrations. Even after incubation for 4 hours under the "immersion" condition, the composition of Example 3 did not visually reduce cell viability at any of the administered concentrations.

[0183] Under the "ALI" condition, three different cell lines (Calu-3, A549, and HAELVi cells) were examined three times at different time points (5 hours later, 2 days later, and 7 days later). After 4 hours, no cytotoxicity was observed in any of the cells, and even after 2 days in the case of Calu-3 cells. In A549 cells, some toxicity was observed after 2 days and 7 days, and in Calu-3 cells, some toxicity was observed after 7 days. Due to technical limitations, quantification of dead cells was not possible. Under the "ALI" condition, three different cell lines (Calu-3, A549, and HAELVi cells) were examined three times at different time points (5 hours later, 2 days later, and 7 days later). After 4 hours, no cytotoxicity was observed in any of the cells, and even after 2 days in the case of Calu-3 cells. In A549 cells, some toxicity was observed after 2 days and 7 days, and in Calu-3 cells, some toxicity was observed after 7 days. Due to technical limitations, quantification of dead cells was not possible. Under the "ALI" condition, three different cell lines (Calu-3, A549, and HAELVi cells) were examined three times at different time points (5 hours later, 2 days later, and 7 days later). After 4 hours, no cytotoxicity was observed in any of the cells, and even after 2 days in the case of Calu-3 cells. In A549 cells, some toxicity was observed after 2 days and 7 days, and in Calu-3 cells, some toxicity was observed after 7 days. Due to technical limitations, quantification of dead cells was not possible. Under the "ALI" condition, three different cell lines (Calu-3, A549, and HAELVi cells) were examined three times at different time points (5 hours later, 2 days later, and 7 days later). After 4 hours, no cytotoxicity was observed in any of the cells, and even after 2 days in the case of Calu-3 cells. In A549 cells, some toxicity was observed after 2 days and 7 days, and in Calu-3 cells, some toxicity was observed after 7 days. Due to technical limitations, quantification of dead cells was not possible. Under the "ALI" condition, three different cell lines (Calu-3, A549, and HAELVi cells) were examined three times at different time points (5 hours later, 2 days later, and 7 days later). After 4 hours, no cytotoxicity was observed in any of the cells, and even after 2 days in the case of Calu-3 cells. In A549 cells, some toxicity was observed after 2 days and 7 days, and in Calu-3 cells, some toxicity was observed after 7 days. Due to technical limitations, quantification of dead cells was not possible. Under the "ALI" condition, three different cell lines (Calu-3, A549, and HAELVi cells) were examined three times at different time points (5 hours later, 2 days later, and 7 days later). After 4 hours, no cytotoxicity was observed in any of the cells, and even after 2 days in the case of Calu-3 cells. In A549 cells, some toxicity was observed after 2 days and 7 days, and in Calu-3 cells, some toxicity was observed after 7 days. Due to technical limitations, quantification of dead cells was not possible.

[0184] Regarding uptake by macrophages, differentiated THP-1 cells were diluted 1:200 in HB Incubated with SS for 4 hours to determine the cell viability of macrophages after exposure. Implementation The composition of Example 3 did not induce cell death, but it was definitely shown that crophagimin was taken up by macrophages.

[0185] Regarding TEER measurement, Calu-3 cells were exposed to HBSS or the composition of Example 3 at three concentrations for 4 hours, and TEER measurement values were extracted at various time points during exposure. If the TEER decreased by ≧50% compared to the control at any given time point, it was considered that the integrity of the barrier function was significantly reduced. When Calu-3 cells were exposed to the composition of Example 3, no effect on the integrity of the barrier function was observed 1 hour after exposure. When exposed at 20 mg / ml, it significantly decreased (i.e., ≧50%) after 2 hours. At a concentration of 10 mg / ml, a slight decrease (i.e., 25 - 35%) was observed at any time point after 2 hours. When exposed at 2 mg / ml, no decrease in the barrier function was observed throughout the test period.

[0186] When Calu-3 cells were exposed to the composition of Example 3, no effect on the integrity of the barrier function was observed 1 hour after exposure. When exposed at 20 mg / ml, it significantly decreased (i.e., ≧50%) after 2 hours. At a concentration of 10 mg / ml, a slight decrease (i.e., 25 - 35%) was observed at any time point after 2 hours. When exposed at 2 mg / ml, no decrease in the barrier function was observed throughout the test period.

[0187] Inflammatory cytokine production The positive control LPS showed the expected behavior in this model. No significant change in cytokines was observed at any time point when the composition of Example 3 was investigated.

[0188] The results are shown in Table 9.

[0189]

Table 9

[0190] In vivo safety and tolerance Six - to eight - week - old Balb / C female mice were administered a total of 3 times at intervals of one day. The mice were given ​​​​​​​​The compositions of Example 1 were administered at 10.0, 5.01, and 2.51 mg / kg. The compositions were administered intratracheally (IT) as an aerosol in an amount of 35 μl / animal using a Microsprayer (registered trademark). After the injection, the mice were observed at 10 minutes, 1, 2, and 4 hours after administration, and then observed daily.

[0191] Table 10 shows the overall observations after administration. "BAR" indicates that the animals were bright , active, and responsive. This is shown in the table.

[0192] [Table 10] [Table 11]

[0193] These data indicate that there were no statistically significant changes in body weight over the three-day treatment period. These results indicate that the compositions of the present invention show sufficient tolerance at the doses used in the tests.

[0194] ​

Claims

1. A pharmaceutical composition comprising: (a) a therapeutically effective amount of clofazimine or a pharmaceutically acceptable derivative or salt thereof; (b) a nonionic surfactant having a hydrophilic-lipophilic balance value exceeding 10; (c) an aqueous liquid selected from water, isotonic saline, buffered saline, and an aqueous electrolyte solution as a carrier; wherein the clofazimine or a pharmaceutically acceptable derivative or salt thereof is provided in the form of particles in a suspension, and the particles of clofazimine or a pharmaceutically acceptable derivative or salt thereof have a median diameter of less than 5 μm and a D90 of less than 6 μm, a pharmaceutical composition.

2. The particles of clofazimine or a pharmaceutically acceptable derivative or salt thereof have an average diameter of less than 2 μm and a D90 of less than 3 μm, the pharmaceutical composition according to claim 1.

3. A pharmaceutical composition comprising: (a) a therapeutically effective amount of clofazimine; (b) a nonionic surfactant having a hydrophilic-lipophilic balance value exceeding 10; (c) an aqueous liquid selected from water, isotonic saline, buffered saline, and an aqueous electrolyte solution as a carrier; wherein the clofazimine is provided in the form of particles in a suspension, and the particles of clofazimine have a median diameter of less than 5 μm and a D90 of less than 6 μm a pharmaceutical composition.

4. The particles have a median diameter of less than 2 μm and a D90 of less than 3 μm, according to claim 3 the pharmaceutical composition described.

5. The clofazimine is provided in one or more polymorphs selected from triclinic FI type, monoclinic FII type, and orthorhombic FIII type , and mixtures of this kind of type, claim 3 or 4 of the pharmaceutical composition described.

6. The clofazimine is provided substantially in orthorhombic FIII type, according to claim 5 the pharmaceutical composition described.

7. The nonionic surfactant is polysorbate 20, polysorbate 60, polysorbate 80, stearyl alcohol, a polyethylene glycol derivative of hydrogenated castor oil having a hydrophilic-lipophilic balance value of 14-16, a polyethylene glycol derivative of hydrogenated castor oil having a hydrophilic-lipophilic balance value of 15-17 , sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, polyoxyethylene (20) oleyl ether, polyoxyethylene (20) cetyl ether, polyoxyethylene (10) cet ​ ​ Tetrahydrofuran, polyoxyethylene (10) oleyl ether, polyoxyethylene (1 00) stearyl ether, polyoxyethylene (10) stearyl ether, polyoxy ethylene (20) stearyl ether, polyoxyethylene (4) lauryl ether, polyoxyethylene (20) cetyl ether, polyoxyethylene (2) cetyl ether , caprylocapryloyl polyoxyl-8 glyceride, polyethylene glycol (20) mo nostearate, polyethylene glycol (40) stearate, polyethylene glycol l (100) stearate, polyethylene glycol (8) stearate, and polyoxy sil 40 stearate, and a mixture thereof, according to any one of claims 1 to 6 The pharmaceutical composition according to item.

8. The nonionic surfactant is polysorbate 80, and The aqueous liquid carrier is distilled water, hypertonic saline, or isotonic saline, according to any one of claims 1 to 7. The pharmaceutical composition according to item.

9. The hypertonic saline is 1% to 7% (w / v) sodium chloride, according to claim 8 The pharmaceutical composition according to item.

10. The nonionic surfactant is ultra-high purity polysorbate 80, and the aqueous liquid carrier Is isotonic saline, according to claim 8. The pharmaceutical composition according to item.

11. The weight osmolality of the composition is in the range of 200 to 700 mOsm / kg, according to claim 1 to 10. The pharmaceutical composition according to any one of items.

12. The weight osmolality of the composition is in the range of 300 to 400 mOsm / kg, according to claim 1 to 10. The pharmaceutical composition according to any one of items.

13. The nonionic surfactant is in the range of 0.001% to 5% (v / v) of the total composition And The amount of clofazimine is in the range of 0.1% to 20% (w / v) of the total composition, according to claim 1 to 10. The pharmaceutical composition according to any one of items.

14. The following steps: (1) Homogenizing a suspension of clofazimine, the nonionic surfactant, and water to obtain a suspension containing clofazimine having an appropriate particle size; (2) Adjusting the pH of the suspension obtained in (1) to a pH between pH 5.5 and pH 7.5; (3) Adjusting the concentration of sodium chloride to an appropriate concentration; (4) Adjusting the weight osmolality to an appropriate level; ​ ​ The pharmaceutical composition according to any one of claims 1 to 13, prepared by a process comprising

15. The following steps: (1) Homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension containing clofazimine with an appropriate particle size; (2) Isolating the clofazimine; (3) Adding the clofazimine to the nonionic surfactant and water; (4) Adjusting the pH of the suspension obtained from (3) to a pH between pH 5.5 and pH 7.5; (5) Adjusting the sodium chloride concentration to an appropriate concentration. The pharmaceutical composition according to any one of claims 1 to 13, prepared by a process comprising

16. The following steps: (1) Micronizing clofazimine to obtain clofazimine with an appropriate particle size; (2) Adding the clofazimine to the nonionic surfactant and water; (3) Adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5; (4) Adjusting the sodium chloride concentration to an appropriate concentration. The pharmaceutical composition according to any one of claims 1 to 13, prepared by a process comprising

17. A suspension in which clofazimine is suspended in water containing the nonionic surfactant and an appropriate concentration of sodium chloride and adjusted to a pH between pH 5.5 and pH 7 .5, prepared by a process comprising homogenizing to obtain clofazimine with an appropriate particle size. The pharmaceutical composition according to any one of claims 1 to 13.

18. The pH is adjusted to 7.4, and the sodium chloride concentration is adjusted to 154 mM sodium chloride. The pharmaceutical composition according to any one of claims 14, 15, or 16.

19. The pH is 7.4, and the appropriate concentration of sodium chloride is 154 mM sodium chloride. The pharmaceutical composition according to claim 17.

20. The micronization of the clofazimine is performed by jet milling, spray drying, ball milling, or supercritical fluid treatment. The pharmaceutical composition according to claim 16.

21. The homogenization in step (1) is performed by high-pressure homogenization, high-shear homogenization, wet milling, ultrasonic homogenization, or a combination of this type of treatment. The pharmaceutical composition according to any one of claims 14, 15, 17, or 19.

22. The homogenization of clofazimine is carried out by multi-stage homogenization, the pharmaceutical composition according to any one of claims 14, 15, 17, 18, 19, or 21.

23. The micronization of clofazimine is carried out by multi-stage micronization, the pharmaceutical composition according to claim 16 or 20 thereof.

24. The clofazimine with the appropriate particle size has an average diameter of less than 5 μm and D90 is particles less than 6 μm, the pharmaceutical composition according to any one of claims 14 to 23.

25. The clofazimine with the appropriate particle size has an average diameter of less than 2 μm and D90 is particles less than 3 μm, the pharmaceutical composition according to any one of claims 14 to 23.

26. The composition according to any one of claims 1 to 25 is aerosolized by a nebulizer selected from ultrasonic nebulizers, charged spray nebulizers, diaphragm nebulizers, jet nebulizers, and mechanical soft mist metered-dose inhalers to prepare a combination pharmaceutical in the form of an inhalation aerosol, wherein the aerodynamic mass median diameter of the particles of the aerosol generated by the nebulizer is 1 to 5 μm, the combination pharmaceutical.

27. The inhalation aerosol is for deposition in the lower respiratory system, the combination pharmaceutical according to claim 26.

28. Atomized 4 to 7% hypertonic saline, metaperiodate, sodium dodecyl sulfate, sodium bicarbonate, tromethamine, silver nanoparticles, bismuth-thiol, ethylenediamine tetraacetic acid, gentamicin-loaded phosphatidylcholine-modified gold nanoparticles, chelating agents, cis -2-decenoic acid, D-amino acids, peptides containing D-amino acid residues, gallium mesoporphyrin IX, gallium protoporphyrin IX, curcumin, patulin, penicillic acid, baicalein, naringenin, ursolic acid, asiatic acid, corosolic acid, fatty acids, host defense peptides, and antimicrobial peptides, agents for dispersing and / or or destroying biofilms, mucolytics and / or mucoactive agents, and / or agents for reducing biofilm formation, the pharmaceutical composition according to any one of claims 1 to 25 for use in combination with.

29. Atomized 4 to 7% hypertonic saline, metaperiodate, sodium dodecyl sulfate, ​ ​ Sodium bicarbonate, tromethamine, silver nanoparticles, bismuth-thiol, ethylenediaminetetraacetic acid, gentamicin-loaded phosphatidylcholine-modified gold nanoparticles, chelating agent, cis-2-decenoic acid, D-amino acid, peptide containing D-amino acid residue, gallium mesoporphyrin IX, gallium protoporphyrin IX, curcumin, patulin, penicillic acid, baicalein, naringenin, ursolic acid, asiatic acid, corosolic acid, fatty acid, host defense peptide, and antimicrobial peptide, an agent for dispersing and / or destroying biofilm, a mucolytic agent and / or a mucoactive agent, and / or a combination with an agent for reducing biofilm formation, the combination medicament according to claim 26 or 27

30. The pharmaceutical composition according to any one of claims 1 to 25 for use in the treatment and / or prevention of lung infections caused by Mycobacteria or other Gram-positive bacteria

31. The combination medicament according to claim 26 or 27 for use in the treatment and / or prevention of lung infections caused by Mycobacteria or other Gram-positive bacteria

32. The pharmaceutical composition according to claim 30, wherein the infection is caused by a bacterial species belonging to the genus Mycobacterium selected from the group of non-tuberculous mycobacteria and Mycobacterium tuberculosis (Mycobacterium tuberculosis) and combinations thereof

33. The combination medicament for use according to claim 31, wherein the infection is caused by a bacterial species belonging to the genus Mycobacterium selected from the group of non-tuberculous mycobacteria and Mycobacterium tuberculosis (Mycobacterium tuberculosis) and combinations thereof

34. The non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and Mycobacterium leprae ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ (Mycobacterium leprae)), and selected from these combinations The pharmaceutical composition for use according to claim 32.

35. The non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium avium), Mycobacterium intracellulare, Mycobacterium intracellulare m intracellulare), Mycobacterium abscessus, and Mycobacterium acterium abscessus), and Mycobacterium leprae (Mycobacterium leprae)), and selected from these combinations The combined medicine for use according to claim 33.

36. The infectious disease is a opportunistic infectious disease selected from pulmonary MAC disease and non-tuberculous mycobacteriosis in patients with cystic fibrosis, chronic obstructive pulmonary disease, or acquired immunodeficiency syndrome. The pharmaceutical composition for use according to claim 32.

37. The infectious disease is a opportunistic infectious disease selected from pulmonary MAC disease and non-tuberculous mycobacteriosis in patients with cystic fibrosis, chronic obstructive pulmonary disease, or acquired immunodeficiency syndrome. The combined medicine for use according to claim 33.

38. The infectious disease is an opportunistic infectious disease of non-tuberculous mycobacteria in patients with cystic fibrosis. The pharmaceutical composition for use according to claim 36.

39. The infectious disease is an opportunistic infectious disease of non-tuberculous mycobacteria in patients with cystic fibrosis. The combined medicine for use according to claim 37.

40. A system for use in imparting antibacterial activity in treating or preventing pulmonary infections caused by mycobacteria or other Gram-positive bacteria, the system comprising: (a)A therapeutically effective dose of clofazimine; (b)A nonionic surfactant having a hydrophilic-lipophilic balance value exceeding 10; (c)An aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solutions １） A nebulized combined medicine containing, 2) A nebulizer, Including, The clofazimine is present in the form of a suspension, And The aerosol particles produced by the system have an aerodynamic mass median diameter of 1 to 5 μm.

41. The nebulizer has a spraying rate of 0.1 to 1.0 ml / min. Use according to any one of claims 28 to 39.

42. 。 The total inhalation volume is between 1 ml and 5 ml. Use according to any one of claims 28 to 39 or 41.

43. ​ ​ ​ ​ The composition is administered before, simultaneously with, or after administration of an agent selected from bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazinamide, rifampicin, moxifloxacin, levofloxacin, and para-aminosalicylate, and mixtures thereof, for use according to any one of claims 28, 30, 32, 34, 36, or 38.

44. The combination is administered before, simultaneously with, or after administration of an agent selected from bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazinamide, rifampicin, moxifloxacin, levofloxacin, and para-aminosalicylate, and mixtures thereof, for use according to any one of claims 29, 31, 33, 35, 37, or 39.

45. The pharmaceutical composition according to any one of claims 1 to 25 for use in the treatment and / or prevention of pulmonary fungal infections or Clostridium difficile or a combination thereof.

46. The combination medicament according to any one of claims 26, 27, 29, 31, 33, 35, 37, or 39 for use in the treatment and / or prevention of pulmonary fungal infections or Clostridium difficile or a combination thereof.

47. The pharmaceutical composition according to claim 45 for use in the treatment and / or prevention of pulmonary fungal infections.

48. The combination medicament according to claim 46 for use in the treatment and / or prevention of pulmonary fungal infections.

49. The pulmonary fungal infection is Candida albicans or Aspergillus fumigatus or a combination thereof, the pharmaceutical composition according to claim 45 or 47.

50. The pulmonary fungal infection is Candida albicans 。 or Aspergillus fumigatus or a combination thereof, the combination medicament according to claim 46 or 48.

51. The composition is administered before, simultaneously with, or after administration of an agent selected from bedaquiline or a pharmaceutically acceptable salt or derivative thereof and amikacin and mixtures thereof, ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ The use according to claim 43.

52. The use according to claim 43, wherein the composition is administered before, simultaneously with, or after administration of bedaquiline or a pharmaceutically acceptable salt or derivative thereof.

53. A method for treating or preventing pulmonary infections in a patient in need thereof, comprising administering the composition according to any one of claims 1 to 25 by inhalation.

54. The method for treatment or prevention according to claim 53, wherein the infection is caused by a mycobacterium species selected from the group of non-tuberculous mycobacteria and mycobacterium tuberculosis (Mycobacterium tuberculosis) and combinations thereof.

55. The method for treatment or prevention according to claim 54, wherein the non-tuberculous mycobacteria are selected from Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, and leprosy bacteria (Mycobacterium leprae), and combinations thereof.

56. The method for treatment or prevention according to claim 53, wherein the infection is an opportunistic infection selected from pulmonary MAC disease and non-tuberculous mycobacteriosis in patients with cystic fibrosis, chronic obstructive pulmonary disease, or acquired immunodeficiency syndrome.

57. The method for treatment or prevention according to claim 56, wherein the infection is an opportunistic infection of non-tuberculous mycobacteria in patients with cystic fibrosis.

58. A method for treating or preventing pulmonary infections caused by mycobacteria or other gram-positive bacteria in a patient in need thereof, comprising administering the composition according to any one of claims 1 to 25 by inhalation before, simultaneously with, or after administration of an agent selected from salts of bedaquiline or a pharmaceutically acceptable derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazinamide, rifampicin, moxifloxacin, levofloxacin, and para-aminosalicylate, and mixtures thereof.

59. The method for treatment or prevention according to claim 58, wherein the agent is bedaquiline or amikacin.

60. The treatment or prevention method according to claim 59, wherein the agent is bedaquiline.

61. The following steps: (1) Homogenizing a suspension of clofazimine, the non-ionic surfactant, and water to obtain a suspension containing clofazimine having an appropriate particle size; (2) Adjusting the pH of the suspension obtained from (1) to a pH between pH 5.5 and pH 7.5; (3) Adjusting the sodium chloride concentration to an appropriate concentration; (4) Adjusting the weight osmolarity to an appropriate level; A process for preparing a pharmaceutical composition according to any one of claims 1 to 13, comprising the above steps.

62. The following steps: (1) Homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension containing clofazimine having an appropriate particle size; (2) Isolating the clofazimine; (3) Adding the clofazimine to the non-ionic surfactant and water; (4) Adjusting the pH of the suspension obtained from (3) to a pH between pH 5.5 and pH 7.5; A process for preparing a pharmaceutical composition according to any one of claims 1 to 13, comprising the above steps.

63. The following steps: (1) Micronizing clofazimine to obtain clofazimine having an appropriate particle size; (2) Adding the clofazimine to the non-ionic surfactant and water; (3) Adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5; (4) Adjusting the sodium chloride concentration to an appropriate concentration; A process for preparing a pharmaceutical composition according to any one of claims 1 to 13, comprising the above steps.

64. A process for preparing a pharmaceutical composition according to any one of claims 1 to 13, comprising homogenizing a suspension in which clofazimine is suspended in water containing the non-ionic surfactant and an appropriate concentration of sodium chloride and having a pH adjusted to a pH between pH 5.5 and pH 7.5 to obtain clofazimine having an appropriate particle size.

65. The process according to any one of claims 61, 62, or 63, wherein the pH is adjusted to 7.4 and the sodium chloride concentration is adjusted to 154 mM sodium chloride.

66. ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ The pH is 7.4 and the appropriate concentration of sodium chloride is 154 mM sodium chloride, the process according to claim 64.

67. The micronization of the clofazimine is carried out by jet milling, spray drying, ball milling, or supercritical fluid treatment, the process according to claim 63.

68. The homogenization in step (1) is carried out by high-pressure homogenization, wet milling, ultrasonic homogenization, or a combination of this kind of treatment, the process according to any one of claims 61, 62, 64, or 66.

69. The homogenization of the clofazimine is carried out by multi-stage homogenization, the process according to any one of claims 61, 62, 64, 65, 66, or 68.

70. The micronization of the clofazimine is carried out by multi-stage micronization, the process according to claim 63 or 20

71. The clofazimine with the appropriate particle size has an average diameter of less than 5 μm and a D90 of less than 6 μm particles, the process according to any one of claims 61 to 70.

72. The clofazimine with the appropriate particle size has an average diameter of less than 2 μm and a D90 of less than 3 μm particles, the process according to any one of claims 61 to 70.

73. The following steps: (a) homogenizing a suspension of clofazimine, the non-ionic surfactant, and water to obtain a suspension containing clofazimine with an appropriate particle size; ( b) adjusting the pH of the obtained suspension to a pH between pH 5.5 and pH 7.5; ( b) adjusting the pH of the obtained suspension to a pH between pH 5.5 and pH 7.5; ( c) adjusting the sodium chloride concentration to an appropriate concentration; and (d) adjusting the weight osmolarity to an appropriate level, steps (b), (c), and (d) can be carried out in the order of (b), (c), (d); (b), (d), (c); (c), (b), (d); (c) , (d), (b); (d), (b), (c); or (d), (c), (b) for preparing the pharmaceutical composition according to any one of claims 1 to 13.

74. The following steps: (a) homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension containing clofazimine with an appropriate particle size; (b) isolating the clofazimine; (c) adding the clofazimine to the non-ionic surfactant and water ​ ​ The step of adding to; (d) adjusting the pH of the resulting suspension to a pH between pH 5.5 and pH 7.5; The step of adjusting the sodium chloride concentration to an appropriate concentration; Including, steps (d) and (e) can be carried out in the order of (d), (e); or (e), (d), for preparing the pharmaceutical composition according to any one of claims 1 to 13 Process.

75. The following steps: (a) micronizing clofazimine to obtain clofazimine with an appropriate particle size; And (b) adding the clofazimine to water containing the nonionic surfactant and an appropriate concentration of sodium chloride and having a pH adjusted to between pH 5.5 and 7.5, for preparing the pharmaceutical composition according to any one of claims 1 to 13 Process. ​ ​ ​

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