JP2021535124A - The composition of clofazimine, the combination containing it, the process for preparing it, the use and method containing it. - Google Patents

Abstract

The present invention relates to an inhalation pharmaceutical composition comprising a therapeutically effective dose of clofazimine, wherein clofazimine is provided in the form of a suspension, and a process for preparing the inhalation pharmaceutical composition. Further, the present invention provides a combination drug containing clofazimine in the form of an aerosol for transpulmonary inhalation. 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.

Description

This application claims priority to US Provisional Application No. 62 / 722,048 filed on August 23, 2018, which is also filed on January 25, 2019. It also claims the priority of US Provisional Application No. 62/769, 322, and the contents of both specifications are incorporated herein by reference.

The present invention is an inhalation pharmaceutical composition comprising a therapeutically effective dose of clofazimine, wherein clofazimine is provided in the form of a suspension; a process for preparing it; as well as the same. Including use and treatment methods. Further, the present invention provides a combination drug containing clofazimine in the form of an aerosol for transpulmonary inhalation.

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.

Clofazimine is a highly hydrophobic (LogP = 7.66) antibiotic liminophenazine with anti-mycobacterial and anti-inflammatory activity, first described in 1957. The structural formula is shown below.

The exact mechanism by which clofazimine exerts its antibacterial activity is unknown. However, it is known to inhibit DNA replication and cell proliferation by preferentially binding to mycobacterial DNA. Other proposed mechanisms of action include membrane damage / destabilization, membrane destabilizing lysophospholipid production, potassium transport and / or disruption of the intracellular redox cycle. In vitro, clofazimine has excellent activity against tubercle bacilli (Mycobacterium tuberculosis) (MTB) including multidrug-resistant strains, but until recently, it has been effective in the treatment of pulmonary tuberculosis. The general idea was that it was not (see, for example, Choro Met al., J Antimiclob Chemother, 2012 Feb, 67 (2): 290-8).

Clofajimin is one of the three major drugs recommended by the World Health Organization for the treatment of leprosy caused by Mycobacterium leprae. In recent years, it has been increasingly used for the treatment of other mycobacterial infections such as drug-resistant tuberculosis caused by nontuberculous mycobacteria (NTM).

Clofazimine is classified as a Class II drug of the Biopharmaceutics Classification System (BCS) because it is hardly soluble in water and exhibits high membrane permeability.

In optimizing the oral dosage form, with the aim of overcoming the problems associated with low oral absorption of drugs and low bioavailability, pulverization, nanonization, and re-using supercritical fluids were used. Various strategies have been used, such as supercritical fluid re-crystallation, spray freeze-drying in liquid, solid dispersions, and solid solutions.

Since clofazimine is classified as a BCS Class II drug, it is usually considered to be an ideal option to be formulated in solid dispersions to improve oral bioavailability (eg, Bushnure et al). . IJRPC 2014, 4 (4), 906-918).

Accordingly, lipophilic clofazimine is usually administered as a polycrystalline suspension in an oil-wax base for the purpose of improving oral absorption. There is a considerable difference in the rate of absorption by the human body after oral administration (45-62%). The adverse effects of clofazimine are dose-related and primarily affect the skin, eyes and gastrointestinal tract, with QT prolongation side effects accompanied by reddish-brown discoloration of the skin and conjunctiva, which is reversible and slowly recovers with drug suspension. Is shown. This is due to the long-term retention throughout the body.

The genus Mycobacterium is a genus of the phylum Actinobacteria and is an independent family of the family Mycobacterium. Mycobacteria have a rod-like shape and a waxy adventitia.

As such, mycobacteria can be divided into three groups:
-Mycobacterium tuberculosis group-causative agent of tuberculosis-Mycobacterium leprae-causative agent of Hansen's disease-Nontuberculous Includes all mycobacteria except mycobacterium (NTM) -tuberculous (M. tuberculosis) and Mycobacterium leprae (M. leprae). Mycobacterium avium complex (MABSC), Mycobacterium avium complex (MAC), etc.

Mycobacterium tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis complexx bacteria. TB is one of the oldest documented human infectious agents and remains a significant contributor to increased mortality and morbidity worldwide. The number of new TB infections in 2015 is estimated to be 10.4 million, with 1.4 million dying from active TB (see, for example, the World Health Organization (WHO) Global Tuberculosis Report 2016). ). In addition to these high prevalence and mortality rates, there is growing concern about the emergence of multidrug-resistant tuberculosis (MDR-TB), and 580,000 patients will be infected with drug-resistant TB in 2015. It is shown that he was doing. Co-morbidity with human immunodeficiency virus (HIV) and the like complicates treatment and was involved in 1.2 million TB cases in 2015.

To treat multidrug-resistant (MDR) infections, WHO has recommended a 9-12 month treatment regimen with second-line anti-TB drugs. In such regimens, such as Bangladesh regimen for 9-12 months, MDR-TB is treated with gatifloxane, ethambutol, pyrazineamide, and clofazimine in combination, with 87.9% of patients absent. Relapsed healing was achieved (eg, Sotguiu, G, et al., "Applicability of the shorter'Bangladesh regimen'in high multidrug-resistant tuberculosisstation

Other studies investigating shortening the duration of TB treatment have shown that clofazimine has no clinical utility after 2 weeks of oral administration (eg, Diacon, HA, et al., " Bactericidal Activity of Pyrazinamide and Clofazimine Alone and in Combinations with Pretomanid and Bedaquiline ”, American Journament Since the theory has been established that this drug binds to serum proteins in the circulating blood with high affinity, it was considered that this lack of efficacy was due to the low bioavailability of the drug. Although clofazimine has been empirically demonstrated to be effective in the treatment of MDR-TB and hyperdrug-resistant TB (XDR-TB), in short-term treatment, due to its low bioavailability after systemic administration. Biological activity seems to be limited (eg, Swanson, R.V., et al., "Pharmacokinetics and Pharmacodynamics of Clofazimine in a Mouse Model of Tuberculosis, Tuberculosis", See -3051).

Treatment of lung infections by inhalation of antibiotics results in higher drug levels in the lungs and reduced side effects compared to systemic delivery (eg, Tow, DJ, et al., "Inhalation". Of antibiotics in cytotic fibrosis ”, European Respiratory Journal (1995), 8, 1594-1604), as a result of which is known to be more bioactive and effective (eg, Hickey, A.J.,". See Inhaled drug treatment for tuberculosis: Past drug and future projects ”, Journal of Controlled Release, (2016), 240, 127-134). Aerosolized administration of clofazimine to a TB infection model significantly improves the removal rate of bacillus bacteria (bacilli) only 28 days after the start of treatment compared to oral administration of clofazimine in vivo mice. Model-proven (eg, Verma, RK, et al., "Inhaled microparticulis contacting clofazimine are efficacious in treatment of experimental Tuberculosis10 See 1052). This short-term improvement in efficacy may be due to higher levels of clofazimine in lung macrophages of tuberculous granulomatosis as a result of direct delivery of clofazimine to the site of infection in the lung.

Therefore, aerosolizing and administering clofazimine to patients infected with MDR TB or XDR-TB should further improve the outcome of the patient's treatment and may shorten the duration of the current treatment regimen.

The nontuberculous mycobacteria (NTM) group, formerly called atypical or ubiquitous mycobacteria, contains more than 150 bacterial species. NTMs are widespread and diverse in the natural environment. These may be detected not only in soil, ground and drinking water, but also in foods such as pasteurized milk and cheese. NTMs are usually considered to be less pathogenic. However, they can cause serious illness in humans, especially those with weakened immunity and those already suffering from lung disease. Currently, NTMs are classified according to their growth rate and are divided into slow-growing (SGM) and fast-growing (RGM) mycobacteria.

The slow-growing Mycobacterium avium complex (MAC) includes Mycobacterium avium, Mycobacterium avium, and Mycobacterium avium intramycobacterium. Intracellulare) strains are included, these are the most important and most commonly found pathogenic NTMs. Mycobacterium kansasii, Mycobacterium malmoense, Mycobacterium xenopi, Mycobacterium mycobacterium, Mycobacterium mycobacterium, Mycobacterium mycobacterium. ), Mycobacterium gordonae, Mycobacterium fortuitum, and Mycobacterium chelonae, most of which also cause lung infections. Mycobacterium marinum is involved in skin and soft tissue infections such as aquarium granuloma.

In particular, RGM causes serious, life-threatening chronic lung disease and is involved in disseminated, often fatal infections. Infection usually results from contaminated material and invasive procedures involving catheters, non-sterile surgical procedures or injections, and foreign body transplants. Contact with shower heads and jacuzzis has also been reported to be at risk of infection. NTM usually causes opportunistic infections in patients with chronic lung disease such as chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF) and in other patients with impaired immune function.

In recent years, rapid-growing (RGM) Mycobacterium abscess (Mycobacterium abscess subspecies Mycobacterium abscess (M. abscess) (M. abscess) (M. abscess). Mycobacterium abscess Boleti, Mycobacterium abscess Bolletii., And Mycobacterium abscess Massiliense, Mycobacterium abscess M. Its importance as a pathogen is becoming apparent, with a much higher lethality rate than any other RGM.

Infection of CF patients with Mycobacterium abscess is particularly problematic, as lung destruction spreads, is often untreatable, and has a high treatment failure rate of 60-66%. (For example, Obregon-Henao A et al, Nontuberculous Agents and Chemotherapy, November 2015, Vol 59, No 11, p. 6904-6912; Qvist, T., Pressler, T. See “Shifting paradigms of nontuberculous mycobacteria in cyclic fibrosis”, Respiratory Research (2014), 15 (1): pp. 41-47).

The link between human infection with NTM and the outbreak of human acquired immunity deficiency syndrome is increasing. Mycobacteria belonging to the Mycobacterium avium complex (MAC) have been identified as the major causative agent of opportunistic infections in patients infected with the human immunodeficiency virus (HIV).

Some strains of NTM are known to form biofilms. A biofilm is a microcolony of bacteria encapsulated in extracellular matrix that imparts stability and resistance to the human immune system. Recently, it has been shown that several species of NTM form biofilms that increase resistance to disinfectants and antibacterial agents. Biofilm assembly progresses through several stages, including reversible attachment, irreversible attachment, biofilm formation by bacterial aggregation, organization, and signal transduction, and final dispersion. During this process, bacteria build a matrix containing extracellular polymer substances (EPS) such as polysaccharides, lipids, and nucleic acids to form complex three-dimensional structures (eg, Sousa S. et al. , International Journal of Mycobacterialogic 4 (2015), 36-43). Specifically, since mycobacteria do not produce exopolysaccharides, the EPS of mycobacteria is different from other biofilms (eg, Zambrano MM, Kolter R. Mycobacterial biofilms: a grayy way to hold it). See Cell.200). Mycobacterial biofilms vary from species to species but may include mycolic acid, glycopeptide lipids, mycolyl-diacylglycerols, lipooligosaccharides, lipopeptides, and extracellular DNA (Rose SJ, Babrak LM, Bermudes LE (2015)). Mycobacterium avium Possible Peptides Extracellular DNA the Tributes to Biofilm Formation, Structural Integrity, and Toralance to Antibiotics. Aggregates in biofilms are known to increase resistance to antibacterial agents (see, eg, Faria S. et al., Journal of Pathogens, Vol 2015, Article ID 809014).

As a new approach to the treatment of NTM lung infections, delivery of aerosolized liposome amikacin / inhalation of amikacin solution sprayed by a jet nebulizer (Rose S. et al, 2014, PLoS ONE, Volume 9, Issue 9, e108703 and In addition to Oliver K. et al, Ann Am Torac Soc Vol 11, No. 1, pp. 30-35), inhalation of dry powder fine particles for transpulmonary delivery of anti-TB drugs (Cholo M et al., J Antimiclob Chemother. 2012 Feb; 67 (2): 290-8 and Fourie B. and Netty O., 2015 Inhalation Magazine, Verma 2013 Antimiclob Agents Chemother) has been proposed.

In the treatment of NTM lung disease, parenteral aminoglycosides, tigecycline, and other promising oral antibiotics such as linezolid, delamanid, and bedaquiline, and, in selected cases, following initial treatment with surgical intervention, amikacin. The multidrug therapy regimen by inhalation has been shown to be promising (Lu Ryu et al., Tuberc Respir Dis 2016; 79: 74-84). However, due to the increasing incidence and prevalence of NTM infections, especially NTM lung disease, and limited treatment options, the bioavailability of currently used antibacterial agents such as clofazimine. There is a need to develop new dosage forms / pharmaceutical formulations that improve. Inhalation may increase efficacy and reduce side effects compared to oral and parenteral treatments.

The combined use of clofazimine and amicacin has been shown in vitro to act synergistically with both Mycobacterium avium and Mycobacterium avium (eg, van Ingen, van Ingen, etc.). J., et al., "In ViroSynergy bestween Clofazimine and Amikacin in Treatment of Mycobacterium Mycobacterium Disease", Antimicrobial63a12 (Atimicrobial63A. Furthermore, synergistic effects of the combined use of clofadimine and bedaquiline against tubercle bacilli (Mycobacterium tuberculosis) have also been shown (eg, Cocol, M. et al., "Efficient Measurement and factoriation". of high-order drug interventions in Mycobacterium tuberculosis ”, Sciences Advances 2017: 3: e170881,11 October 2017). In addition, a synergistic effect of the combined use of clofazimine / bedaquiline has been shown against the nontuberculous mycobacteria Mycobacterium absessus (Ruth, MM et al., "A Bedaquiline / Clofazimine Combination". Regimen Might Add Activity to the Treatment of Clinically Relevant Non-Tuberculous Mycobacterium.

Pathogenic fungi are becoming more prominent as a major cause of human mortality. Current estimates suggest that deaths from deep-seated fungal diseases are comparable to the more well-known infectious diseases such as tuberculosis. Candida albicans, Cryptococcus neoformans, and Aspergillis fumigatus are the most common pathogenic fungi in humans. Each of these strains is involved in the infection of hundreds of thousands of people each year, but mortality is unacceptably high due to inadequate diagnostic methods and limited treatment options. Clofazimine has been shown to be effective against multiple fungi as a combination (eg, Robbins, N., et al., "An Antifungal Combination Matrix Ideas a Rich Pool of Adjuvant Molecules". Pathogens ”, Cell Reports 13, 1481-1492, November 17, 2015). The fungus also acts as a commensal, engraftment, and / or pathogen in cystic fibrosis (eg, Photomall, SH and McElvaney, NG., "Fungi in the cytotic fibrosis". rung: Bystanders or pathogens? ”, The International Journal of Biochemistry & Cell Biology 52 (2014), 161-173).

Clofazimine has low oral bioavailability and high drug resistance due to its low water solubility. A special solubilizing and stabilizing drug for formulation in a liquid aqueous carrier for aerosolization and the like by a nebulizer for the purpose of depositing aerosol particles in the lower lung. Techniques are also needed.

According to one embodiment of the invention:
(A) With therapeutically effective doses of clofazimine or pharmaceutically acceptable derivatives or salts thereof;
(B) With a nonionic surfactant having a hydrophilic-lipophilic balance value of more than 10;
(C) With an aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solution;
A pharmaceutical composition containing
Clofazimine or a pharmaceutically acceptable derivative or salt thereof is provided in the form of particles in a suspension.
And a pharmaceutical composition is provided in which 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.

According to another embodiment of the 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.

In another embodiment of the invention:
(A) With therapeutically effective doses of clofazimine;
(B) With a nonionic surfactant having a hydrophilic-lipophilic balance value of more than 10;
(C) With an aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solution;
A pharmaceutical composition containing
Clofazimine is provided in the form of particles in suspension,
And the clofazimine particles are provided with a pharmaceutical composition having a median diameter of less than 5 μm and a D90 of less than 6 μm.

In another embodiment of the invention, the clofazimine particles have a median diameter of less than 2 μm and a D90 of less than 3 μm.

The compositions of the invention are aerosolized with a suitable nebulizer into the alveoli of the aerosolized lower respiratory system (ie, bronchi, bronchioles, and central and lower peripheral lungs). ) Is greatly improved, thereby substantially improving the therapeutic effect.

More preferably, the inhaler should be further adapted to locally deliver an aerosol with an optimal particle size distribution to the lungs for the purpose of uniform deposition in the lower respiratory system.

Accordingly, the present invention provides aerosols with particle sizes that facilitate delivery to the alveoli and bronchioles. Suitable aerodynamic particle sizes targeting the alveoli and bronchioles are between 1-5 μm. Larger particles selectively deposit in the upper lungs, i.e., the bronchi and trachea and the mouth and pharynx, i.e., the oropharyngeal region. Therefore, the inhaler is adapted to produce aerosols with aerodynamic mass media 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.

The present invention allows the deposition of active agents in the lower (ie, deep) respiratory system by aerosolly administering clofazimine to the lungs in the form of a suspension, thereby making the BCS highly hydrophobic. It is based on the unexpected finding that the bioavailability of Class II agonists is improved, resulting in significantly improved therapeutic efficacy and reduced systemic side effects.

In other embodiments, this finding results in opportunistic infections of infections caused by mycobacteria and gram-positive bacteria, particularly lung infections with NTMs, such as CF, COPD, and patients with impaired immune function (such as HIV patients). Antibacterial drug treatment such as is improved.

Furthermore, the present invention aims not only to eliminate systemic side effects of established oral treatment regimens for lung infections caused by gram-positive bacteria, especially TB and NTM infections in the lung, but also to reduce the dose and duration of treatment of clofazimin. do.

It will be appreciated by those skilled in the art that this application will also disclose any combination of the individual features disclosed herein.

Definitions The term "pharmaceutically acceptable salt" refers to a salt that retains the biological efficacy and properties of the compounds of the invention and is not biologically or otherwise undesirable. In many cases, the compounds of the present invention can form salts of acids and / or bases due to the presence of amino and / or carboxyl groups or similar groups. Pharmaceutically acceptable acid addition salts can be formed using inorganic and organic acids. Examples of the inorganic acid capable of inducing a salt include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Organic acids that can induce salts include, for example, acetic acid, propionic acid, naphthoic acid, oleic acid, palmitic acid, pamo (embon) acid, stearic acid, glycolic acid, pyruvate, oxalic acid, maleic acid, malon. Acids, succinic acid, fumaric acid, tartaric acid, citric acid, ascorbic acid, glucoheptic acid, glucuronic acid, lactic acid, lactobionic acid, tartaric acid, benzoic acid, cinnamon acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfon Acids, salicylic acids and the like can be mentioned.

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 and the like; ammonium, potassium, sodium, calcium and magnesium. Each salt of is particularly preferable. Examples of the organic base capable of inducing a salt include substituted amines such as primary, secondary and tertiary amines and naturally substituted amines, cyclic amines, basic ion exchange resins and the like, and specific examples thereof. , Isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, histidine, arginine, lysine, venetamine, N-methyl-glucamine, ethanolamine and the like. Other acids include dodecyl sulfate, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, and saccharin.

According to the present invention, it is preferable to use methanesulfonic acid, maleic acid, isonicotinic acid, nicotinic acid, malonic acid, and salicylate, particularly clofazimin methanesulphonate, in addition to the free base.

As used herein, the term "pharmaceutically acceptable derivative" means, for example, a compound disclosed in US Pat. No. 9,540,336, US Pat. No. 9,540, The entire disclosure of specification 336 is incorporated herein by reference as it constitutes part of this specification. In addition to this, the meaning of the derivative is Lu, Y. , Zhen, M. et al. , Wang, B. , Fu, L. , Zhao, W. et al. , Li, P.I. , Xu, J. et al. , Zhu, H. et al. , Jin, H. , Yin, D. , Huang, H. et al. , Upton, AM. and Ma, Z. , "Clofazimine Analogs with Efficacy against experimental Tuberculosis and redened Potential for Acculation" Antimicrobial Agent (11) As described in 5185-5193. Further, the "pharmaceutically acceptable derivative" of the compound is, for example, a prodrug of the above compound. In general, a prodrug is a derivative of the compound capable of providing an active form of the compound after administration. Derivatives of this type 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.

A "therapeutically effective amount", a "therapeutically effective dose", or a "pharmaceutically effective dose", or a "pharmaceutically effective salt thereof" is disclosed in the present invention. Alternatively, it means an amount of the derivative showing a therapeutic effect. The therapeutically effective dose of clofazimine is the therapeutically effective amount. Thus, the therapeutically effective amount as used herein means the amount of clofazimine that provides the desired therapeutic effect, as determined from clinical trial results and / or infection studies using animal models.

The amount and daily dose of clofazimine can be determined mechanically by one of ordinary skill in the art and will vary depending on several factors, eg, the specific microbial strain involved. In addition, this amount may also depend on the patient's height, weight, gender, age, and medical history. The therapeutically effective amount in the case of prophylactic treatment is an amount effective in preventing microbial infection.

"Therapeutic effect" is to reduce one or more symptoms of an infection to some extent, and also includes curing the infection. "Cure" means that the symptoms of active infection are eliminated, and the excess of live bacteria involved in the infection is completely or substantially eliminated and detected by conventional measurements. Includes being below the threshold. However, certain long-term or permanent effects of infection may exist even after healing has been achieved (eg, extensive tissue damage). As used herein, "therapeutic effect" is defined as a statistically significant reduction in host bacterial mass, the development of resistance, or an improvement in infectious symptoms as determined by human clinical outcomes or animal studies. ..

As used herein, "treat," "treatment," or "treating" is the administration of a pharmaceutical composition / combination for prophylactic and / or therapeutic purposes. Point to.

The term "prophylactic treatment" refers to treating a patient who is not yet infected but is susceptible to or otherwise at risk for a particular infection. The term "therapeutic treatment" refers to treating a patient who is already suffering from an infectious disease. Therefore, in a preferred embodiment, the treatment is to administer a therapeutically effective amount of clofazimine (either for therapeutic or prophylactic purposes) to the mammal.

Unless otherwise specified herein, the term "inhalation" means transpulmonary inhalation.

Unless otherwise specified herein, the term "infection (disease)" as used herein means a lung infection (disease).

Unless otherwise stated, the term "substantially" used to refer to the purity of a compound means that the purity of the compound exceeds 95%.

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

Unless otherwise stated, the term "appropriate concentration" refers to the concentration of an ingredient in a composition or combination that provides a pharmaceutically acceptable composition or combination.

Pharmaceutical Compositions and Combinations The following water grades are particularly suitable for the invention: sterile purified water, sterile injection water, sterile water for irrigation, sterile water for. Equivalent to water quality grades in accordance with sterility (USP) and, for example, the European Pharmacopoeia or the National Formula.

The aqueous electrolyte solution used as an aqueous liquid carrier according to the present invention may further contain sodium chloride, potassium chloride, lithium chloride, magnesium chloride, calcium chloride, or a mixture thereof.

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

Clofazimine has been shown to have at least four polymorphisms (eg, Bannigan, et al., "Investigation into the Solid and Solution Properties of Known and Novell Polymorphism Des. 2016, 16 (12), pp. 7240-7250). Clofazimine can be present in triclinic FI, monoclinic FII, and orthorhombic FIII. Another type, FIV, is also found only at high temperatures.

Therefore, in a further embodiment of the invention:
(A) With therapeutically effective doses of clofazimine;
(B) With a nonionic surfactant having a hydrophilic-lipophilic balance value of more than 10;
(C) With an aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solution;
A pharmaceutical composition containing
Clofazimine is provided in the form of particles in suspension,
And the median diameter of the clofazimine particles 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 triclinic FI type. Provided are pharmaceutical compositions provided in one or more polymorphs selected from monoclinic FII and orthorhombic FIII types, as well as mixtures of this type. In other embodiments, clofazimine is substantially provided in orthorhombic type FIII.

In a further embodiment of the invention, a pharmaceutical composition according to any of the composition embodiments described herein is provided, wherein the nonionic surfactant is polysolvate 20 (eg, Tween® 20). , Polysolvate 60 (eg, Tween® 60), Polysorbate 80 (eg, Tween® 80), stearyl alcohol, polyethylene glycol of hydrogenated castor oil having a hydrophilic-lipophilic balance value of 14-16. Derivatives (eg, Cremophor® RH40), polyethylene glycol derivatives of hydrogenated castor oil having a hydrophilic-lipophilic balance value of 15-17 (eg, Cremophor® RH60), sorbitan monolaurate (eg, Cremophor®). , Span® 20), sorbitan monopalmitate (eg, Span® 40), sorbitan monostearate (eg, Span® 60), polyoxyethylene (20) oleyl ether (eg, eg. Brij® 020), polyoxyethylene (20) cetyl ether (eg Brij® 58), polyoxyethylene (10) cetyl ether (eg Brij® C10), polyoxyethylene (eg, Brij®) ( 10) Oleyl ether (eg, Brij® O10), polyoxyethylene (100) stearyl ether (eg, Brij® S100), polyoxyethylene (10) stearyl ether (eg, Brij®). S10), polyoxyethylene (20) stearyl ether (eg, Brij® S20), polyoxyethylene (4) lauryl ether (eg, Brij® L4), polyoxyethylene (20) cetyl ether (eg, Brij®) ( For example, Brij® 93), polyoxyethylene (2) cetyl ether (eg Brij® S2), caprilocaproyl polyoxyl-8 glyceride (eg Labrasol®), polyethylene glycol. (20) Steerate (eg, Myrj ™ 49), Polyethylene Glycol (40) Steerate (eg, Myrj ™ S40), Polyethylene Glycol (100) Steerate (eg, Myrj ™ S100), Polyethylene. Glycol (8) stearate (eg, Myrj ™ S8), and polyo It is selected from xyl 40 stearate (eg, Myrj ™ 52), as well as mixtures thereof.

In another embodiment of the 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 another embodiment of the invention, a pharmaceutical composition is provided, the hypertonic saline is 1% -7% (w / v) sodium chloride. In a further embodiment of the invention, a pharmaceutical composition is provided, the nonionic surfactant is ultra-high purity polysorbate 80 (eg, NOF Corporation Polysorbate 80 (Hx2)), and the aqueous liquid carrier is isotonic saline. It is a saline solution.

In another embodiment of the invention, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, wherein the weight osmolal concentration of the composition is in the range of 200-700 mOsm / kg. In a further embodiment, the weight osmolal concentration of the composition is in the range of 300-400 mOsm / kg.

In a further embodiment of the invention, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, the nonionic surfactant being 0.001% to 5% of the total composition. It is in the range of% (v / v) and the amount of clofazimin is in the range of 0.1% to 20% (w / v) of the whole composition.

In another embodiment of the invention, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, wherein the pharmaceutical composition is described in the following step:
(1) A step of homogenizing a suspension of clofazimine, a nonionic surfactant, and water to obtain a suspension containing clofazimine of an appropriate particle size.
(2) A step of adjusting the pH of the suspension obtained from (1) to a pH between pH 5.5 and pH 7.5, and
(3) Steps to adjust the sodium chloride concentration to an appropriate concentration,
(4) Steps to adjust the weight osmolality to an appropriate level,
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 other embodiments, the homogenization of step (1) is performed by high pressure homogenization, high shear homogenization, wet grinding, ultrasonic homogenization, or a combination of this type of treatment. In another embodiment, the homogenization of clofazimine is carried out by multi-step homogenization. In other embodiments, clofazimines of suitable particle size are particles with an average diameter of less than 5 μm and a D90 of less than 6 μm. In a further embodiment, clofazimines of appropriate particle size are particles with an average diameter of less than 2 μm and a D90 of less than 3 μm.

In a further embodiment of the invention, a pharmaceutical composition according to any one of the composition embodiments described herein is provided, wherein the pharmaceutical composition is described in the following step:
(1) A step of homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension containing clofazimine of an appropriate particle size.
(2) Steps to isolate clofazimine and
(3) The step of adding clofazimine to the nonionic surfactant and water, and
(4) The step of adjusting the pH of the suspension obtained from (3) to a pH between pH 5.5 and pH 7.5, and
(5) Steps to adjust the sodium chloride concentration to an appropriate concentration,
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 in step (1) is performed by high pressure homogenization, high shear homogenization, wet grinding, ultrasonic homogenization, or a combination of this type of treatment. In other embodiments, the homogenization of clofazimine is carried out by multi-step homogenization. In other embodiments, clofazimines with suitable particle sizes are particles with an average diameter of less than 5 μm and a D90 of less than 6 μm. In a further embodiment, clofazimine having an appropriate particle size is a particle having an average diameter of less than 2 μm and 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, the composition comprising:
(1) A step of pulverizing clofazimine to obtain clofazimine having an appropriate particle size, and
(2) The step of adding clofazimine to the nonionic surfactant and water, and
(3) A step of adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5, and
(4) Steps to adjust the sodium chloride concentration to an appropriate concentration,
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 other embodiments, the pulverization of clofazimine is carried out by jet mill milling, spray drying, ball mill milling, or supercritical fluid treatment. In other embodiments, clofazimine atomization is performed by multi-step atomization. In other embodiments, clofazimines of suitable particle size are particles with an average diameter of less than 5 μm and a D90 of less than 6 μm. In a further embodiment, clofazimines of appropriate particle size are particles with an average diameter of less than 2 μm and 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, which comprises clofazimine, a nonionic surfactant, chloride at an appropriate concentration. Prepared by a process comprising homogenizing a suspension containing sodium and suspended in water whose pH was adjusted between pH 5.5 and pH 7.5 to obtain clofazimine of appropriate particle size. To. 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, homogenization is carried out by high pressure homogenization, high shear homogenization, wet grinding, ultrasonic homogenization, or a combination of this type of treatment. In other embodiments, the homogenization of clofazimine is carried out by multi-step homogenization. In other embodiments, clofazimines of suitable particle size have an average particle size of less than 5 μm and a D90 of less than 6 μm. In a further embodiment, clofazimines of appropriate particle size are particles with an average diameter of less than 2 μm and a D90 of less than 3 μm.

In another embodiment, it is a process for preparing a pharmaceutical composition according to any of the composition embodiments described herein, wherein the following steps:
(1) A step of homogenizing a suspension of clofazimine, a nonionic surfactant, and water to obtain a suspension containing clofazimine of an appropriate particle size.
(2) A step of adjusting the pH of the suspension obtained from (1) to a pH between pH 5.5 and dpH 7.5, and
(3) Steps to adjust the sodium chloride concentration to an appropriate concentration,
(4) Steps to adjust the weight osmolality to an appropriate level,
A process is provided that includes.

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, homogenization is carried out by a combination of high pressure homogenization, wet grinding, ultrasonic homogenization, or this type of treatment. In a further embodiment, the homogenization of clofazimine is carried out by multi-step homogenization. In a further embodiment, clofazimines of suitable particle size are particles with an average diameter of less than 5 μm and a D90 of less than 6 μm. In other embodiments, clofazimines of suitable particle size are particles with an average diameter of 2 μm and D90 of less than 3 μm.

In other embodiments, it is a process for preparing embodiments of the pharmaceutical compositions described herein.
(1) A step of homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension containing clofazimine of an appropriate particle size.
(2) The step of isolating clofazimine and
(3) The step of adding clofazimine to the nonionic surfactant and water, and
(4) A step of adjusting the pH of the suspension obtained from (3) to a pH between pH 5.5 and pH 7.5, and
(5) Steps to adjust the sodium chloride concentration to an appropriate concentration,
A process is provided that includes.

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, homogenization is carried out by a combination of high pressure homogenization, wet grinding, ultrasonic homogenization, or this type of treatment. In a further embodiment, the homogenization of clofazimine is carried out by multi-step homogenization. In a further embodiment, clofazimines of suitable particle size are particles with an average diameter of less than 5 μm and a D90 of less than 6 μm. In other embodiments, clofazimines of suitable particle size are particles with an average diameter of 2 μm and a D90 of less than 3 μm.

In a further embodiment, it is a process for preparing a pharmaceutical composition according to any one of the pharmaceutical composition embodiments described herein, wherein the following steps:
(1) A step of pulverizing clofazimine to obtain clofazimine having an appropriate particle size, and
(2) The step of adding clofazimine to the nonionic surfactant and water, and
(3) A step of adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5, and
(4) Steps to adjust the sodium chloride concentration to an appropriate concentration,
A process is provided that includes.

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 pulverization of clofazimine is carried out by jet mill milling, spray drying, ball mill milling, or supercritical fluid treatment. In a further embodiment, clofazimine atomization is carried out by multi-step atomization. In a further embodiment, clofazimines of suitable particle size are particles with an average diameter of less than 5 μm and a D90 of less than 6 μm. In other embodiments, clofazimines of suitable particle size are particles with an average diameter of 2 μm and a D90 of less than 3 μm.

In other embodiments, a process for preparing a pharmaceutical composition according to any one of the pharmaceutical composition embodiments described herein, wherein clofazimine is a nonionic surfactant, at an appropriate concentration. A process comprising homogenizing a suspension containing sodium chloride and suspended in water whose pH was adjusted between pH 5.5 and pH 7.5 to obtain clofazimine of appropriate particle size. 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, homogenization is carried out by a combination of high pressure homogenization, wet grinding, ultrasonic homogenization, or this type of treatment. In a further embodiment, the homogenization of clofazimine is carried out by multi-step homogenization. In a further embodiment, clofazimines of suitable particle size are particles with an average diameter of less than 5 μm and a D90 of less than 6 μm. In other embodiments, clofazimines of suitable particle size are particles with an average diameter of 2 μm and 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, wherein the following steps: (a) chloride, nonionic surfactant. The step of homogenizing the suspension of the agent and water to obtain a suspension containing chloride of appropriate particle size; (b) pH of the resulting suspension, pH 5.5-5. Including a step of adjusting the pH between 5; (c) adjusting the sodium chloride concentration to an appropriate concentration, and (d) adjusting the weight osmolality concentration to an appropriate level; step (b). , (C) and (d) are (b), (c), (d); (b), (d), (c); (c), (b), (d); (c), A process is provided that can be performed in the order (d), (b); (d), (b), (c); or (d), (c), (b).

In another embodiment, a process for preparing a pharmaceutical composition according to any one of the composition embodiments described herein, wherein the following steps: (a) Clofazimine and a suspension of a non-aqueous liquid. A step of homogenizing the turbidity to obtain a suspension containing clofazimine of appropriate particle size; (b) a step of isolating clofazimine; (c) the clofazimine in a nonionic detergent and water. The step of adding; (d) adjusting the pH of the obtained suspension to a pH between pH 5.5 and pH 7.5; and (e) adjusting the concentration of sodium chloride to an appropriate concentration. A process is provided in which steps (d) and (e) can be performed in the order of (d), (e); or (e), (d).

In another embodiment, it is a process for preparing a pharmaceutical composition according to any one of the composition embodiments described herein, wherein the following steps: (a) clofazimine, suitable granules. The step of atomizing to obtain clofazimine of diameter and (b) clofazimine containing a nonionic surfactant, an appropriate concentration of sodium chloride, the pH was adjusted between pH 5.5-7.5. A process comprising adding to water is provided.

In another embodiment of the present invention, a combination drug in the form of an inhalation aerosol, wherein the composition according to any one of the composition embodiments described herein is an ultrasonic nebulizer, a charged spray type. A combination drug prepared by aerosolization with a nebulizing device selected from an ejectron spray, a vibrating membrane nebulizer, a jet nebulizer, and a mechanical soft mist metering inhaler is provided by the nebulizer. The aerosol particles produced have an aerodynamic mass median diameter of 1-5 μm. In a further embodiment, the inhalation aerosol is for depositing in the lower respiratory system. In another embodiment, the ejection speed of the atomizer is 0.1 to 1.0 ml / min. In other embodiments, the total inhalation volume is between 1 ml and 5 ml.

In other embodiments, the pharmaceutical composition according to any one of the composition embodiments described herein is a sprayed 4-7% hypertonic physiological saline, metaperiodate, dodecyl. Sodium sulfate, sodium hydrogen carbonate, tromethamine, silver nanoparticles, bismuth-thiol, ethylenediamine tetraacetic acid, gentamycin-supported phosphatidylcholine-modified gold nanoparticles, chelating agent, cis-2-decenopartic acid. Amino acid, peptide containing D-amino acid residue (D-enantiometric peptide), gallium mesoporphyrin IX, gallium protoporphyllin IX, curcumin, patulin, penicillic acid, baicalene, naringenin, ursolic acid, asiatic acid, corosolic acid, fatty acid, host Used in combination with agents selected from protective peptides and antimicrobial peptides to disperse and / or destroy biofilms, mucolytic agents and / or mucilage activators, and / or agents that reduce biofilm formation. A pharmaceutical composition for this purpose is provided. In other embodiments, the composition for this use is bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazineamide, rifampine, moxifloxacin, levofloxacin. , And para-aminosalicylate, and agents selected from mixtures thereof, are administered before, simultaneously, or after.

In another embodiment, a combination drug according to any of the combination embodiments described herein, atomized 4-7% hypertonic physiological saline, metaperiodate, sodium dodecyl sulfate, hydrogen carbonate. Sodium, tromethamine, silver nanoparticles, bismus-thiol, ethylenediamine tetraacetic acid, gentamicin-supported phosphatidylcholine-modified gold nanoparticles, chelating agent, cis-2-decenoic acid, D-amino acid, peptide containing D-amino acid residues, gallium mesoporphyrin Disperses and / or disrupts biofilms selected from IX, gallium protoporphyrin IX, curcumin, patulin, penicillic acid, baicaline, naringenin, ursoric acid, aciatic acid, corosolic acid, amino acids, host defense peptides, and antimicrobial peptides. A combination drug for use in combination with an agent for, a mucolytic agent and / or a mucilage activator, and / or an agent for reducing biofilm formation is provided. In other embodiments, the combination for this use is bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazineamide, rifampine, moxifloxacin, levofloxacin, And para-aminosalicylate, and agents selected from mixtures thereof, are used to administer the compositions of the invention before, at the same time, or after. 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, pharmaceutical compositions according to any one of the composition embodiments described herein are used for the treatment and / or prevention of lung infections caused by mycobacteria or other Gram-positive bacteria. Provided to do. In a further embodiment, the infection is a group of nontuberculous mycobacteria and tubercle bacilli (Mycobacterium tuberculosis) and the genus Mycobacterium selected from a combination thereof. It is caused by the bacterial species belonging to. In a further embodiment, the nontuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium avium, and Mycobacterium avium. (Mycobacterium leprae), as well as a combination thereof. In other embodiments, the infection is opportunistic selected from pulmonary MAC disease and nontuberculous infection in patients with cystic fibrosis, chronic obstructive lung, or acquired immunodeficiency syndrome. It is an infectious disease. In another embodiment, the infection is an opportunistic infection with nontuberculous mycobacteria in a patient with cystic fibrosis. In other embodiments, the composition for this use is bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazineamide, rifampine, moxifloxacin, levofloxacin. , And para-aminosalicylate, and agents selected from mixtures thereof, are administered before, simultaneously, or after. 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 combinatorial drug according to any of the combinatorial embodiments described herein is provided for use in the treatment and / or prevention of lung infections caused by mycobacteria or other Gram-positive bacteria. Will be done. In a further embodiment, the infectious agent is selected from nontuberculous mycobacteria and tubercle bacilli (Mycobacterium tuberculosis) and combinations thereof, the genus Mycobacterium. It is caused by the bacterial species belonging to. In a further embodiment, the non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium avium, and Mycobacterium avium. It is selected from Mycobacterium avium, as well as combinations thereof. In another embodiment, the infection is an opportunistic infection selected from pulmonary MAC disease and nontuberculous mycobacteriosis in patients with cystic fibrosis, chronic obstructive lung disease, or acquired immunodeficiency syndrome. .. In another embodiment, the infection is an opportunistic infection with nontuberculous mycobacteria in a patient with cystic fibrosis. In other embodiments, the combination for this use is bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazineamide, rifampine, moxifloxacin, levofloxacin, And para-aminosalicylate, and agents selected from mixtures thereof, are used to administer the compositions of the invention before, at the same time, or after. In other embodiments, the combination for this use is the invention 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. Is used to administer the composition of. In other embodiments, the combination for this use is used to administer the composition of the invention before, simultaneously with, or after administration of bedaquiline or a pharmaceutically acceptable salt or derivative thereof.

In other embodiments, a system is provided for use in imparting antibacterial activity in treating or preventing lung infections caused by mycobacteria or other Gram-positive bacteria, which system is:
1)
(A) With therapeutically effective doses of clofazimine;
(B) With a nonionic surfactant having a hydrophilic-lipophilic balance value of more than 10;
(C) With an aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solution;
With atomized combination medicines, including
2) Atomizer and
Including
Clofazimine exists in the form of a suspension,
The aerosol particles produced by this system have an aerodynamic mass center diameter of 1 to 5 μm.

In a further embodiment, any one of the composition embodiments described herein for use in the treatment and / or prevention of pulmonary fungal infections or Clostridium difficile or combinations thereof. The pharmaceutical composition according to the above is provided. In other embodiments, pharmaceutical compositions according to any one of the composition embodiments described herein are provided for use in the treatment and / or prevention of pulmonary fungal infections. In a further embodiment, the pulmonary fungal infection is Candida albicans or Aspergillus fumigatus or a combination thereof.

In a further embodiment, a combination pharmaceutically according to any one of the combination embodiments described herein for use in the treatment and / or prevention of pulmonary fungal infections or Clostridium difficile or combinations thereof is provided. Will be done. Provided are combinatorial formulations according to any one of the combinatorial embodiments described herein for use in the treatment and / or prevention of pulmonary fungal infections. In a further embodiment, the pulmonary fungal infection is Candida albicans or Aspergillus fumigatus, or a combination thereof.

In another embodiment, a method of treating or preventing a lung infection in a patient in need thereof, administered by inhalation of a composition according to any one of the composition embodiments described herein. Methods are provided, including doing. In other embodiments, the infection is selected from the nontuberculous mycobacteria and tubercle bacilli (Mycobacterium tuberculosis) groups and combinations thereof, the genus mycobacterium. ) Is caused by the bacterial species belonging to. In a further embodiment, the non-tuberculous mycobacteria are Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium avium, and Mycobacterium avium. It is selected from Mycobacterium avium, as well as combinations thereof. In a further embodiment, 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. be. In another embodiment, the infection is an opportunistic infection with nontuberculous mycobacteria in a patient with cystic fibrosis.

In a further embodiment, a method of treating or preventing a lung infection caused by mycobacteria or other gram-positive bacteria in a patient in need thereof, a salt of bedaquiline or a pharmaceutically acceptable derivative thereof. , Cefoxitin, amikacin, clarithromycin, pyrazineamide, rifampine, moxifloxacin, levofloxacin, and para-aminosalicylate, and agents selected from a mixture thereof before, simultaneously, or after administration. , A method comprising inhaling a composition according to any one of the composition embodiments described herein is provided. In other embodiments, the agent is bedaquiline or amikacin. In a further embodiment, the agent is bedaquiline.

Particle size and distribution The therapeutic effect of aerosol therapy depends on the dose dosed and its distribution. Aerosol particle size is one of the important variables that determine the amount and distribution of drug aerosols in the lungs.

In general, inhaled aerosol particles tend to be deposited by one of two mechanisms: collision (usually dominating larger aerosol particles) and sedimentation (dominant with smaller aerosol particles). Collisions occur when the momentum of the inhaled aerosol particles is large enough that the particles do not follow the air flow, resulting in a physiological surface. In contrast, sedimentation occurs when very small aerosol particles carried along the inspiratory flow reach a physiological surface as a result of sedimentation, primarily due to the action of gravity in the lower respiratory system.

Drug delivery to the lungs can be done by inhaling the aerosol through the oropharynx. Aerosol particles with aerodynamic diameters greater than about 5 μm generally do not reach the lungs. Instead, they tend to hit the back of the throat and can be swallowed and orally absorbed. Aerosol particles with a diameter of about 3 to about 5 μm are small enough to reach the upper-to-mid-plumony region (guided airway), but 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 smaller than about 0.5 μm in diameter tend to be expelled with breathing during resting breathing, but can also be deposited in the alveolar region by breath-holding.

Statistical descriptors are used because the aerosols used for pulmonary drug delivery are composed of a wide range of aerosol particle sizes. Aerosols used for pulmonary drug delivery are usually expressed in their mass median diameter (MMD). That is, half of the mass contained in the aerosol particles is larger than that of MMD, and half of the mass contained in the aerosol particles is smaller than that of MMD. If the particle density is uniform, the volume center diameter (VMD) can be used interchangeably with MMD. The VMD and MMD determinations are made using laser diffraction. The width of the distribution is expressed in geometric standard deviation (GSD). On the other hand, the deposition of aerosol particles in the airway can be expressed more accurately by using the aerodynamic diameter of the particles, so the central diameter of the aerodynamic mass is usually used. MMAD measurements are made by inertial collision or flight time measurements. For water particles, the VMD, MMD, and MMAD should be equal. However, if the humidity is not controlled while the aerosol passes through the impactor, dehydration will occur and the MMAD readings will be smaller than the MMD and VMD. In order to use this description, measurements of VMD, MMD, and MMAD are considered to be performed under controlled conditions so that the description of VMD, MMD, and MMAD is comparable. ..

However, for purposes of explanation, the aerosol particle size of the aerosol particles shall be measured at room temperature using the Next Generation Impactor (NGI) in accordance with the United States Pharmacopeia Convention. (In Process Revision <601> Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers, Pharmacopoeia , Mark Nagal "Particle Size Analysis of Aerosols from Medical Internals", KONA Powder and Particle Journal (2004), Volume 22, page-65.

According to the present invention, the particle size of the aerosol is optimized for maximal deposition of clofazimine at the site of infection and for maximum tolerability. The aerosol particle size can be expressed in terms of aerodynamic mass median diameter (MMAD). Large particles (eg, MMAD> 5 μm) tend to deposit extrathoracically and in the upper airway because they are too large to travel along the flexion of the airway. Deposition of large particles in the upper respiratory tract can lead to intolerance symptoms (eg, cough and bronchospasm).

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

However, by guiding breathing techniques, larger particles can pass through the extrathoracic cavity and upper respiratory tract, allowing them to penetrate deeper into the lungs than during resting breathing, thereby in the aerosol. And deposition in the lower respiratory system will increase. The breathing method can be slowed down to 100 ml / min by instruction. Therefore, when using the instructed breathing method, the preferred MMAD of the aerosol should be less than about 10 μm.

Another similarly important factor (other than the aerosol particle size) is the particle size and particle size distribution of the solid particles, in this case clofazimine particle size and distribution. The particle size of a solid particle of a given aerosol particle must be smaller than the aerosol particle containing it. Larger aerosol particles may contain one or more solid particles. Moreover, when dealing with dilute suspensions, the majority of aerosol particles may be free of solid particles.

Therefore, it is desirable to use solid drug particles that are significantly smaller than the MMAD of aerosol particles. For example, when the MMAD of aerosol particles is 3 μm, it is desirable that the solid particles are 1 μm or less.

Another consideration is, for example, when using a vibrating mesh nebulizer, where the pharmaceutical product is delivered through the pores of the plate, which breaks down the suspension into droplets. In that case, for this reason, it is also necessary to make the solid particles smaller than the pores so that they can pass through the pores.

The particle size of the solid in the suspension can be given by the average diameter of the particles or by the distribution of the particles. The D90 value indicates that 90% of the particles in the suspension are below their average diameter.

Atomizer For aqueous and other non-pressurized liquid systems, various atomizers, including small volume nebulizers, are available for aerosolizing the formulations. The compressor-driven sprayer incorporates jet spray technology, which uses compressed air to produce a liquid aerosol. This type of instrument is described, for example, in Healthdyne Technologies, Inc. Invacare, Inc. Mountain Medical Equipment, Inc. Pari Recovery, Inc. Mada Medical, Inc. Puritan-Bennet; Schuco, Inc. , DeVilvis Health Care, Inc. ; And commercially available from Hospitak, Inc. Ultrasonic nebulizers utilize mechanical energy in the form of vibrating piezoelectric crystals to generate inhalable droplets, such as Omron Heathare, Inc. And DeVilvis Health Care, Inc. Vibrating mesh nebulizers utilize either piezoelectric or mechanical pulses to generate inhalable droplets. Other examples of atomizers used with clofazimin described herein are US Pat. No. 4,268,460; US Pat. No. 4,253,468; US Pat. No. 4,046; , 146; US Pat. No. 3,286,255; US Pat. No. 4,649,911; US Pat. No. 4,510,929; US Pat. No. 4,624,251; No.; US Pat. No. 5,164,740; US Pat. No. 5,586,550; US Pat. No. 5,758,637; US Pat. No. 6,644,304; Book; US Pat. No. 6,338,443; US Pat. No. 5,906,202; US Pat. No. 5,934,272; US Pat. No. 5,960,792; US Pat. No. 5,971,951; US Pat. No. 6,070,575; US Pat. No. 6,192,876; US Pat. No. 6,230,706; US Pat. No. 6,230,706; 6,349,719; US patent 6,367,470; US patent 6,543,442; US patent 6,584,971; US patent 6. , 601,581; US Pat. No. 4,263,907; US Pat. No. 5,709,202; US Pat. No. 5,823,179; US Pat. No. 6,192. , 876; US Pat. No. 6,644,304; US Pat. No. 5,549,102; US Pat. No. 6,083,922; US Pat. No. 6,161,536; No.; US Pat. No. 6,264,922; US Pat. No. 6,557,549; and US Pat. No. 6,612,303; all of these. The contents are incorporated herein by reference as they form part of this specification. Examples of commercially available nebulizers that can be used with the clofazimine compositions described herein include Respirgard II®, Aeroneb®, Aeroneb® Pro, and Aeroneb, manufactured by Aerogen. Registered Trademarks) Go; AERx® and AERx Essens ™, manufactured by Aradigm; I-neb Respironics, Inc. Porta-Neb®, Freeway Freedom®, Sidestream, Ventstream; and PARI, GmbH PARI LCPlus®, PARI LC-Star®, and e-Flow7m. Other non-limiting examples are disclosed in US Pat. No. 6,196,219.

According to the invention, the pharmaceutical composition is preferably aerosolized using a spray device selected from an ultrasonic nebulizer, a charged spray nebulizer, a vibrating membrane nebulizer, a jet nebulizer, or a mechanical soft mist metered dose inhaler. can do.

It is preferred that the device controls the patient's inhalation rate either electrically or mechanically.

In a further preferred embodiment, the generation of the aerosol by the instrument is actuated by the inhalation action of the patient, like the AKITA device.

Preferred (commercially available) examples of the above-mentioned atomizers / appliances used in accordance with the present invention are Vector Fox, Pari eFlow, Pari Trek S, Philips Innospire mini, Philips InnoSpire Go, Medspray device, Aerone, Aerone. , Akita, Medspray Economy, and Respirat.

Use in Treatment and / or Prevention The pharmaceutical compositions and combination drugs (aerosols, aerosolized formulations) and systems according to the invention are mycobacteria or Staphylococcus aureus (methicillin resistant and bancomycin). Treatment of pulmonary infections caused by other Staphylococcus aureus (including intermediate resistant strains), Streptococcus aureus pneumoniae, and Enterococcus spp. Alternatively, it is intended to be used for prevention. The pharmaceutical composition and combination drug of the present invention can also be used for the treatment and / or prevention of staphylococcus aureus infection.

Dosing of Clofazimine According to the invention, the pharmaceutical composition is delivered by atomizing about 1-5 ml, preferably 1-2 ml of the pharmaceutical composition of the invention.

Therefore, based on the clofazimine concentration in the pharmaceutical composition being about 20 mg / ml, the target fill dose is about 1-5 ml, which corresponds to 20-100 mg of clofazimine.

The daily lung dose (ie, the amount deposited in the lungs) of clofazimine administered in accordance with the present invention is about 5-10 mg in the case of M. abscess infection, which is about 5-10 mg. Corresponds to a nominal dose of 15 to 30 mg (dose administered by the device).

For those skilled in the art, the transpulmonary dose of clofazimine to be administered (and thus the filling / nominal dose / spray volume) is the minimum inhibitory concentration of clofazimine for each bacterial strain well established in the art. It will be understood that it will be adjusted mechanically based on MIC).

Therefore, the daily pulmonary dose will be divided according to the frequency of administration once or twice daily.

According to the present invention, clofazimine is administered once or twice daily so that the total daily lung dose is consequently about 5-10 mg.

It will be apparent to those skilled in the art that the amounts mentioned above relate to the free base of clofazimine and the doses of derivatives and salts should be adjusted based on the MIC of each compound and strain.

Biofilm denaturing agent / biofilm modifying agent
For the purpose of reducing the viscosity of sputum during aerosol treatment and destroying the biofilm present, the treatment and / or prevention according to the invention comprises the additional administration of a mucolytic agent and / or a biofilm destroying agent. Can be done.

These agents can be formulated as a fixed combination or administered simultaneously or subsequently with the clofazimine-containing pharmaceutical composition / aerosol combination according to the invention.

Agents for dispersing / destroying biofilms used in accordance with the present invention, mucolytic agents and / or mucilage activators, and / or agents for reducing biofilm formation are sprayed 4-7% hypertonic saline. Water, metaperiodated acid salt, sodium dodecyl sulfate, sodium hydrogencarbonate, tromethamine, silver nanoparticles, bismus-thiol, ethylenediamine tetraacetic acid, gentamicin-supported phosphatidylcholine-modified gold nanoparticles, chelating agent, cis-2-decenoic acid, D- Amino Acids, Peptides Containing D-Amino Acid Residues, Gallium Mesoporphyrin IX, Gallium Protoporphyllin IX, Curcumin, Patulin, Penicillic Acid, Baicaline, Naringenin, Ursoleic Acid, Aciatic Acid, Corosolic Acid, Fatty Acids, Host Defense Peptides, and Antimicrobials Selected from peptides.

In addition, other pharmaceutically active agents can be used in combination with the pharmaceutical composition / aerosol combination according to the invention. This type of activator is bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazineamide, riffampine, moxifloxacin, levofloxacin, and para-aminosalicylate. , As well as a mixture of these.

These agents can be formulated as a combination drug, or can be administered before, simultaneously with, or after the combination of the pharmaceutical composition / aerosol containing clofazimine according to the present invention.

The embodiments shown below serve to more fully illustrate the modes in which the invention described above is used and to explain the best possible embodiments for carrying out the various aspects of the invention. The embodiments according to the present invention are included in the claims described herein.

Tests The exemplary compositions and combinations shown below were prepared according to the process described herein.

Example 1
200 mg of clofazimine (as triclinic type I), 90 mg of sodium chloride, and 9.5 ml of water were mixed twice with an Ultra-Turrax homogenizer at 10,000 rpm for 5 minutes. 0.5 ml of polysorbate 80 (NOF Hx2) was added. The mixture was treated 7 times (3 minutes each) at 70% amplitude with an ultrasonic probe (Branson Digital Sonifier ™ 250D equipped with a Bandelin Sonoplus Probe MS73). The volume was adjusted to 10 ml with water. The suspension was filtered through a VWR foldable qualitative filter paper (303, particle retention capacity 5-13 μm, size: 150 mm) to give the composition of Example 1. The median particle size of clofazimine in the composition of Example 1 was 3.9 μm, and D90 was 6.7 μm. The concentration of clofazimine was determined to be 7.16 mg / ml by measuring at 280 nm by ultraviolet / visible spectroscopy and calibrating with a 1 mg / ml stock solution of clofazimine diluted in mobile phase.

The composition of Example 1 is shown in Table 1.

Preparation of Orthorhombic Type III Clofazimine A slurry of clofazimine (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 portion of the slurry was collected by filtration through a crucible filter and vacuum dried in an oven at a maximum temperature of 40 ° C. This gave a substantially pure (≧ 98%) orthorhombic type III clofazimine with a yield of 8.64 g.

Example 2
A suspension containing 6 g of orthorhombic type III clofazimine in 100 ml of water containing 0.5% polysorbate 80 (NOF Hx2) and 0.6% sodium chloride using Ultra-Turrax® 10 Preliminarily atomized at 000 rpm for about 40 seconds. Preliminary preparations were prepared by adding water containing 0.6% sodium chloride to a volume of 300 ml. 300 ml of this suspension is charged into the inlet of the homogenizer M-110EH-30 microfluidizer (Microfluidics, Westwood, MA, USA) and the suspension is circulated in the H30Z chamber at 5,000 psi for 15 minutes for prehomogenization. Performed the steps. The second H10Z chamber was then mounted in series with the first chamber and the suspension was homogenized at 25,000 psi for an additional 23 minutes. When the 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. The concentration of clofazimine was determined to be 16.05 mg / ml by measuring at 280 nm by ultraviolet / visible spectroscopy and calibrating with a 1 mg / ml stock solution of clofazimine diluted in the mobile phase.

The composition of Example 2 is shown in Table 2.

Example 3
Suspension of clofazimin (crystal transformation: orthorhombic type III) in a solution of water, sodium chloride, and polysorbate 80 is a suspension of the M-110EH-30 Microfluidizer® Processor (chamber: H30Z and). The composition of Example 3 was produced by using G10Z) in an H30Z-G10Z arrangement and operating at a pressure of 28,250 psi for 30 minutes. The median particle size of the obtained clofazimine particles was 1.28 μm, and D90 was less than 2 μm.

The composition of Example 3 is shown in Table 3.

Viscosity measurement The viscosity of the composition of Example 3 was measured in a stress control mode using a STRESSTECH Rheometer. A hollow gap geometry was used to rotate the spindle continuously to ensure that the particles remained suspended at each temperature point (during temperature points). The viscosity was measured at 20 ° C., 25 ° C., and 30 ° C. with stresses of 0.01, 0.05, and 0.1 Pa, respectively. The average value of the viscosities obtained by performing two separate sample fillings (loading) is shown in Table 4 below.

Animal Models and Efficacy Tests Clinically isolated NTM strains with the aim of obtaining preliminary data to establish concentration levels in lung tissue after direct respiratory delivery rather than systemic administration of clofazimine. The ability of the compositions of the invention to inhibit proliferation was tested in an in vivo acute lung infection mouse model. Two separate mouse models were used to investigate NTM lung infections by target strain. Mycobacterium avium (2285) strain and Mycobacterium avium (Mycobacterium avium) 103 strain were used for the test (details of the strain were registered in the EMBL / GenBank / DDBJ database (2014). (September) "Phylogetic analogysis of Mycobacterium avium genome sexuences". Hazbon M.H., Riojas M.A., Damon A. M. King S., Sohrabi A.). These two strains have already been used as models of NTM infection in the literature (Obregon-Henao et al. 2015 Antimiclob Agents Chemother; and Chan et al. Animal Models of Non-Tuberculosis In-Tuberculosis ).

In vivo safety test in Balb / C mice:
6-8 week old Balb / C female mice were obtained from Charles River for evaluation of in vivo safety and tolerability. Mice were rested for a week before dosing. Three healthy mice were provided for each dose of clofazimine and administered three times every other day. Clofazimine 10.0, 5.01 and 2.51 mg / kg were administered to mice in the composition of Example 1. Three healthy mice were administered an aerosol of this compound intratracheally using a Microsprayer® three times every other day.

The safety of clofazimine has been confirmed to be 20 mg / kg (forced oral administration, 200 μl). 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, intratracheal). Therefore, the composition of Formula I is considered safe at 10.0 mg / kg and is well tolerated.

Determination of Minimum Inhibitor Concentration The Minimum Inhibitor Concentration (MIC) test uses Mueller Hinton (MH) broth (cation-adjusted) (calcium and magnesium ion concentrations recommended by CLSI standard M7-A7) (Becton Dickinson). Then, it was carried out by a microbross dilution method. The MIC test was also performed by a trace solution dilution method using 7H9 broth (Sigma-Aldrich). The rationale for using both MH and 7H9 broths for compound screening is that antimycobacterial compounds have been shown to exhibit different MIC activities depending on the broth used in the MIC test.

M. abscess was grown on a 7H11 agar plate (Sigma-Aldrich) at 35-37 ° C. in the air (in ambient air) (depending on the bacterial strain) for 3 days, and M. avium (M. avium). .Avium) was grown on a 7H11 agar plate (Sigma-Aldrich) in air at 37 ° C. for 21-30 days.

Colony forming units (CFU) were harvested from agar plates and added to either MH broth or 7H9 broth supplemented with 0.05% tween-80 on day 3 (M. abscess) or 12 After the day (M. abium), the optical density (OD) is measured by absorbance, and (OD) 0.08 to 0.1 (McFarland turbidity standard solution No. 0.5) is obtained. It was grown in air at 35-37 ° C. It was then confirmed by preparing the cell suspension in physiological saline to match (OD) 0.08-0.1 (McFarland turbidity standard solution No. 0.5). A stock solution of the compound was prepared by suspending the compound in DMSO at a concentration of 1.28 mg / ml and used immediately with a test range of 64-0.062 μg / ml. Subsequently, 180 μl of broth (either MH or 7H9) was added to the first row of the 96-well plate and 100 μl of broth was added to the remaining row of the 96-well plate. 20 μl of the stock solution of the compound was added to the wells in the first row and serially diluted. Finally, 100 μl of NTM cell suspension was added to all wells except the control well with medium only. QC reagent specific to each microorganism: 1) Bacterial-only negative control 2) Medium-only negative control 3) Clarithromycin-positive drug control.

The OD of M. abscess was measured after 3 days, and the OD of M. avium was measured after 12 days. Following these measurements, plates were measured using the Resazurin Microtiter Assay Plate method. Briefly, this method is the addition of resazurin (7-hydroxy-3H-phenoxazine-3-one-10-oxide) to a 96-well plate. Resazurin is a blue pigment that emits weak fluorescence by itself, but when reduced irreversibly, it becomes resorphin that emits strong pink fluorescence. It has been used as a redox indicator to determine bacterial cell viability in MIC measurements.

The test was conducted in triplets. Test # 1 was performed after storing the composition of Example 1 at 4 ° C. for 2 months, test # 2 was performed after 4 months, and test # 3 was performed after 5 months.

Minimum inhibitory concentration in the presence and absence of CF sputum Measurement of the minimum inhibitory concentration was performed as described above.

To investigate the effect of sputum in patients with cystic fibrosis (CF) on the antibacterial activity of clofazimin (CFZ) and the composition of Example 1, sputum was recovered from patients who had not been treated with antibacterial agents within 48 hours. Then, the sputum was sterilized by exposing it to UV light to remove the internal bacteria. After sterilization, M. abscess, M. avium, M. intracellulare, and M. chimera were incubated in 10% CF sputum. Later, a MIC test was performed. The MIC of the composition of Example 1 was measured in the presence and absence of sputum in patients with cystic fibrosis according to the same CLSI protocol as described above. All tests were conducted in duplicate.

Table 5 shows the MIC values of clofazimine and the composition of Example 1 in the presence and absence of sputum.

From the results shown in Table 5, it can be seen that both clofazimine and the composition of Example 1 show stable MICs against a wide range of nontuberculous mycobacterial species.

These data indicate that the composition of Example 1 exhibits strong in vitro activity against both M. abscess and M. avium and is stable for at least this period. It suggests.

Mouse model of M. abscess in SCID mice 6-8 week old SCID female mice were purchased from Charles River. Mice were rested for a week before infection.

1 ml of a test stock solution (working stock) of 103 strains of M. abscess was taken and stored at −80 ° C. until use. The preparative solution was thawed for infection, disintegrated 20 times using a 1 ml luer lock syringe fitted with a 26 g needle and diluted with sterile 1 x PBS.

Acute infected SCID mouse models were ^{infected with lung by non-invasive intratracheal injection of 1 × 10 6} CFU / animal (M. abscess 103 strain).

Three mice were sacrificed on the first day after infection, and the amount of bacterial uptake (bacterial uptake) was determined. All lungs, spleen, and liver were removed and homogenized in 4.5 ml of 1 x PBS. The homogenizet was serially diluted at a dilution ratio of 1:10, and the diluted solution (0-1-2-3-4-5-6-7) was smeared on a 7H11 agar plate. The flat plate was allowed to stand in a dry air incubator (depending on the strain) at 32 ° C. for 7 days.

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

Mice were sacrificed 2 days after administration of the last dose of compound. Six mice in all groups (no treatment control, clofazimine (forced oral administration), composition of Example 1, and amikacin-treated mice) were sacrificed and the bacterial count was measured. Lung homogenate 0-1-2-3-4-5-6-7, spleen 0-1-2-3-4-5-6-7, and liver 0-1-2-3-4- The plate was smeared with 5-6-7.

A protection value represented by Log 10 of at least 0.60 suggests that the activity is statistically significant. Statistical analysis was performed by first converting CFU to logarithm, then evaluating it with one-way ANOVA, and then performing analysis of variance by multiple comparison with one-way Tukey test (GraphPad Prism analysis software). Differences are considered significant at a 95% confidence level.

_{Table 6 shows the mean value of Log 10} CFU data and the standard error (SEM) of the mean value after infecting SCID mice with M. abssusus. "N" is the total number of animals per group at slaughter.

The data in Table 6 show that treatment with the composition of Example 1 significantly reduced the amount of bacteria recovered from the lungs and spleen of animals infected with M. abscess. .. This decrease in bacterial mass was statistically improved as compared with treatment with amikacin or oral clofazimine.

Mouse model of Beige mice infected with M. avium 6-8 week old female Beige mice were purchased from Charles River. Mice were rested for a week before infection.

Acute infection Beige mouse model to 1 × 10 ⁸ colony forming units (CFU) / ml of (M · avium (M. avium) 2285 strain rough type) non-invasively by lung infection by exposure to an aerosol. 1 ml of M. avium strain 2285 stock solution for testing was taken, frozen, and stored at −80 ° C. until use. The preparative solution was thawed for infection, disintegrated 20 times using a 1 ml luer lock syringe equipped with a 26 g needle, and diluted with sterile 1 x phosphate buffered saline.

Three mice were sacrificed on days 1 and 7 after infection and bacterial uptake was measured. Whole lungs, spleen, and liver were removed, homogenized in 4.5 ml of 1 x PBS and diluted 1:10. Dilute (0-1-2-3-4-5-6-7) is smeared on 7H11 / OADC, TSA, and charcoal agar plates and in a dry air incubator at 32 ° C (depending on the strain) for 30 days. Incubated.

10.0 mg / kg of the composition of Example 1 was administered by the transpulmonary route using Microspirer® (35 μl) and clofazimine (forced oral administration) was administered in an amount of 200 μl per mouse. These were started 7 days after infection and continued every other day for 10 days.

Mice were sacrificed 5 days after administration of the final dose of compound. Six mice in all groups (no treatment control, clofazimine (forced oral administration), and the composition of Example 1) were sacrificed and the bacterial count was measured. Lung homogenate 0-1-2-3-4-5-6-7, spleen 0-1-2-3-4-5-6-7, and liver 0-1-2-3-4- 5-6-7 was smeared on a flat plate.

A defense value represented by Log10 of at least 0.60 suggests that the activity is statistically significant. Statistical analysis was performed by first converting CFU to logarithm, then evaluating it with one-way ANOVA, and then performing ANOVA with multiple comparisons with one-way Tukey test (SigmaStat software program). Differences are considered significant at a 95% confidence level.

Table 7 shows the average value _{of Log 10} CFU data after infecting Beige mice with M. avium.

The data in Table 7 show that treatment with the composition of Example 1 significantly reduced the amount of bacteria recovered from the lungs and spleen of animals infected with M. avium. ..

Chronically infected Beige mouse model 6-8 week old Beige mice were rested for 1 week before infection. On day 0, mice were lung-infected with M. avium 2285rough type 1 × 10 ^{8 CFU.} Three mice were sacrificed on the first day and six mice were sacrificed on the 27th day, and the uptake of the fungus and the amount of the fungus before the treatment were measured. Whole lungs, spleen, and liver were removed, homogenized in 1 x PBS 4.5 ml, and diluted (0-1-2-3-4-5-6-7) on 7H11 agar plates and charcoal agar plates. It was smeared. The flat plate was allowed to stand in a dry air incubator at 37 ° C. for 25 to 30 days.

The remaining infected Beige mice were treated every other day starting from the 28th day, for a total of 14 treatments. Animals were treated with one of the following treatments: saline (Microsprayer®, 35 μl); clofazimine (forced oral administration, 20 mg / kg, 200 μl); composition of Example 1 (IT). , Microspirer®, 10 mg / kg, 35 μl).

Mice were sacrificed on day 57, two days after the last treatment. The flat plate was allowed to stand in a dry air incubator at 37 ° C. for 30 days.

Statistical analysis was performed by first converting CFU to logarithm, then evaluating it with one-way ANOVA, and then performing ANOVA with multiple comparisons with one-way Tukey test. Differences are considered significant at a 95% confidence level.

Table 8 shows the average value _{of Log 10} CFU data after chronic infection of Beige mice with M. avium.

These data suggest that clofazimine is slow to penetrate the granulomatous structures formed by the established "chronic" NTM-infected animal model. The compositions of the present invention do not have this problem and appear to be able to maintain antimycobacterial activity even after the infection has fully settled.

Effect of barrier function on soundness and inflammation after in vitro exposure of the composition of Example 3 to lung epithelial cells Cell viability To evaluate the cell viability of lung epithelial cells Three different types of cells: Calu-3; A549; and hAELVi cells were used under two in vitro conditions. Cells were treated either under "immersion conditions" (ie, in cell culture medium on a Transwell ™ plate) or under "air-liquid interface" simulated conditions (ALI) with the cell culture medium removed from the apical side of the cells. Under "immersion conditions", Calu-3 cells were exposed to the composition of Example 3 at three doses (10%, 50%, or 100%) for 4 hours. To assess cell viability, cells were stained with acridine orange / propidium iodide (AO / PI) staining to distinguish between live and dead cells. Red fluorescence indicates dead cells.

Uptake by macrophages THP-1 cells were incubated with 124 ng / ml of phorbol 12-millistart 13 acetate (PMA) for 3 days to differentiate into macrophage-like cells. Once the cells had matured, they were exposed to the composition of Example 3 (diluted 1: 200 with Hanks balanced salt solution (HBSS)) for 4 hours. Cell viability after exposure was determined by staining cells with AO / PI as described above.

Transepithelial Electrical Resistance (TEER) Measurements Calu-3 cells were ^{seeded on Transwell ™ 3460 to 1 × 10 5} cells / well and allowed to grow for 12 days until confluent was reached. EVOM2 (World Precision Instruments, Friedberg, Germany) was used and TEER measurements were performed according to the manufacturer's instructions. After seeding, Calu-3 cells were exposed to either saline (negative control) or the composition of Example 3 (concentration: 20 mg / ml, 10 mg / ml, or 2 mg / ml). After exposing the cells for 2-4 hours, TEER was measured.

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

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 sample. Separate ELISA kits were used for TNF-α, IL-6, IL-8, and IL-10 according to the manufacturer's instructions.

After performing one-way analysis of variance (ANOVA), statistical analysis was performed using Tukey post-hoc test. If the probability value <0.05, it was judged to be statistically significant.

Results Even after incubation for 4 hours under "immersion" conditions, the composition of Example 3 did not visually reduce cell viability at any concentration administered.

Under "ALI" conditions, 3 different cells (Calu-3, A549, and HAELVi cells) were investigated 3 times (5 hours, 2 days, and 7 days) at different time points. After 4 hours, little or no cytotoxicity was observed in any of the cells, and in the case of Calu-3 cells, after 2 days. Some toxicity was observed in A549 cells after 2 and 7 days, and in Calu-3 cells after 7 days. Due to technical limitations, dead cells could not be quantified.

For macrophage uptake, differentiated THP-1 cells were incubated at 1: 200 HBSS for 4 hours to determine cell viability of the macrophages after exposure. The composition of Example 3 did not induce cell death, but it was certainly shown that clofazimine was taken up by macrophages.

For TEER measurements, Calu-3 cells were exposed to HBSS or the composition of Example 3 at three concentrations for 4 hours, and TEER measurements were extracted at various time points during exposure. At any given time point, a ≧ 50% reduction in TEER compared to the control was considered to be a significant reduction in barrier function integrity.

When Calu-3 cells were exposed to the composition of Example 3, no effect was observed on the soundness of the barrier function even 1 hour after the exposure. Exposure at 20 mg / ml resulted in a significant (ie ≧ 50%) decrease after 2 hours. At a concentration of 10 mg / ml, a slight decrease (ie 25-35%) was seen at any time point after 2 hours. No decrease in barrier function was observed throughout the test period when exposed at 2 mg / ml.

LPS, a positive control for inflammatory cytokine production, behaved as expected in this model. The composition of Example 3 did not show any significant changes in cytokines at any of the time points investigated.

The results are shown in Table 9.

In vivo safety and tolerability 6-8 week old Balb / C female mice were administered every other day for a total of 3 doses. Mice were dosed with the composition of Example 1 at 10.0, 5.01, and 2.51 mg / kg. The composition was administered intratracheally (IT) at an amount of 35 μl / animal of aerosol using a Microsprayer®. After injection, mice were observed 10 minutes, 1, 2, and 4 hours after administration, followed by daily observation.

Table 10 shows a comprehensive observation after administration. "BAR" indicates that the animal was smart, active, and responsive.

These data indicate that body weight did not show a statistically significant change after 3 days of treatment.

These results indicate that the compositions of the invention are well tolerated at the doses used in the study.

Claims (75)

It is a pharmaceutical composition:
(A) With therapeutically effective doses of clofazimine or pharmaceutically acceptable derivatives or salts thereof;
(B) With a nonionic surfactant having a hydrophilic-lipophilic balance value of more than 10;
(C) With an aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solution;
Including
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. The pharmaceutical composition according to claim 1, wherein 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. It is a pharmaceutical composition:
(A) With therapeutically effective doses of clofazimine;
(B) With a nonionic surfactant having a hydrophilic-lipophilic balance value of more than 10;
(C) With an aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solution;
Including
The clofazimine is provided in the form of particles in a suspension and is provided.
And the particles of clofazimine are pharmaceutical compositions having a median diameter of less than 5 μm and a D90 of less than 6 μm. The pharmaceutical composition according to claim 3, wherein the particles have a median diameter of less than 2 μm and a D90 of less than 3 μm. The clofazimin is provided in one or more polymorphs selected from triclinic FI, monoclinic FII, and orthorhombic FIII, and mixtures of this type. The pharmaceutical composition according to claim 3 or 4. The pharmaceutical composition according to claim 5, wherein the clofazimine is provided in substantially orthorhombic type FIII. 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 to 16, and a hydrophilic-oil lipophilic balance value. Polyethylene glycol derivative of hydrogenated castor oil, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, polyoxyethylene (20) oleyl ether, polyoxyethylene (20) cetyl ether, polyoxy Ethylene (10) cetyl ether, polyoxyethylene (10) oleyl ether, polyoxyethylene (100) stearyl ether, polyoxyethylene (10) stearyl ether, polyoxyethylene (20) stearyl ether, polyoxyethylene (4) lauryl Ether, polyoxyethylene (20) cetyl ether, polyoxyethylene (2) cetyl ether, caprilocaproyl polyoxyl-8 glyceride, polyethylene glycol (20) monostearate, polyethylene glycol (40) stearate, polyethylene glycol ( 100) The pharmaceutical composition according to any one of claims 1 to 6, selected from 100) stearate, polyethylene glycol (8) steerate, and polyoxyl 40 steerate, and mixtures thereof. The invention according to any one of claims 1 to 7, wherein the nonionic surfactant is polysorbate 80, and the aqueous liquid carrier is distilled water, hypertonic saline, or isotonic saline. Pharmaceutical composition. The pharmaceutical composition according to claim 8, wherein the hypertonic saline solution is 1% to 7% (w / v) sodium chloride. The pharmaceutical composition according to claim 8, wherein the nonionic surfactant is ultra-high purity polysorbate 80, and the aqueous liquid carrier is isotonic saline. The pharmaceutical composition according to any one of claims 1 to 10, wherein the weight osmolal concentration of the composition is in the range of 200 to 700 mOsm / kg. The pharmaceutical composition according to any one of claims 1 to 10, wherein the weight osmolal concentration of the composition is in the range of 300 to 400 mOsm / kg. The nonionic surfactant ranges from 0.001% to 5% (v / v) of the total composition and the amount of clofazimine is 0.1% to 20% (w / v) of the total composition. The pharmaceutical composition according to any one of claims 1 to 10, which is in the range of v). The following steps:
(1) A step of homogenizing a suspension of clofazimine, the nonionic surfactant, and water to obtain a suspension containing clofazimine of an appropriate particle size.
(2) The step of adjusting the pH of the suspension obtained from (1) to a pH between pH 5.5 and pH 7.5, and
(3) Steps to adjust the concentration of sodium chloride to an appropriate concentration,
(4) Steps to adjust the weight osmolality to an appropriate level,
The pharmaceutical composition according to any one of claims 1 to 13, which is prepared by a process comprising the above. The following steps:
(1) A step of homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension containing clofazimine of an appropriate particle size.
(2) The step of isolating the clofazimine and
(3) The step of adding the clofazimine to the nonionic surfactant and water, and
(4) The step of adjusting the pH of the suspension obtained from (3) to a pH between pH 5.5 and pH 7.5, and
(5) Steps to adjust the sodium chloride concentration to an appropriate concentration,
The pharmaceutical composition according to any one of claims 1 to 13, which is prepared by a process comprising the above. The following steps:
(1) A step of pulverizing clofazimine to obtain clofazimine having an appropriate particle size, and
(2) The step of adding the clofazimine to the nonionic surfactant and water, and
(3) A step of adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5, and
(4) Steps to adjust the sodium chloride concentration to an appropriate concentration,
The pharmaceutical composition according to any one of claims 1 to 13, which is prepared by a process comprising the above. A suspension containing the nonionic surfactant, an appropriate concentration of sodium chloride and having clofazimine suspended in water adjusted to a pH between 5.5 and 7.5, with an appropriate particle size. The pharmaceutical composition according to any one of claims 1 to 13, which is prepared by a process comprising homogenization to obtain clofazimine. The pharmaceutical composition according to any one of claims 14, 15, or 16, wherein the pH is adjusted to 7.4 and the sodium chloride concentration is adjusted to 154 mM sodium chloride. 17. The pharmaceutical composition of claim 17, wherein the pH is 7.4 and the appropriate concentration of sodium chloride is 154 mM sodium chloride. The pharmaceutical composition according to claim 16, wherein the pulverization of clofazimine is performed by jet mill pulverization, spray drying, ball mill pulverization, or supercritical fluid treatment. 13. The pharmaceutical composition according to paragraph 1. The pharmaceutical composition according to any one of claims 14, 15, 17, 18, 19, or 21, wherein the homogenization of clofazimine is performed by multi-step homogenization. The pharmaceutical composition according to claim 16 or 20, wherein the pulverization of clofazimine is carried out by multi-step pulverization. The pharmaceutical composition according to any one of claims 14 to 23, wherein the clofazimine having an appropriate particle size is a particle having an average diameter of less than 5 μm and a D90 of less than 6 μm. The pharmaceutical composition according to any one of claims 14 to 23, wherein the clofazimine having an appropriate particle size is a particle having an average diameter of less than 2 μm and a D90 of less than 3 μm. The composition according to any one of claims 1 to 25 is aerosolized by a spraying device selected from an ultrasonic nebulizer, a charged spray nebulizer, a vibrating membrane nebulizer, a jet nebulizer, and a mechanical soft mist metering inhaler. A combination drug in the form of an inhalation aerosol prepared by
A combination drug having an aerodynamic mass median diameter of the aerosol particles produced by the spraying device of 1-5 μm. The combination drug according to claim 26, wherein the aerosol for inhalation is intended to be deposited in the lower respiratory system. Sprayed 4-7% hypertonic physiological saline, metaperiodide, sodium dodecyl sulfate, sodium hydrogencarbonate, tromethamine, silver nanoparticles, bismuth-thiol, ethylenediamine tetraacetic acid, gentamicin-supported phosphatidylcholine-modified gold nanoparticles, chelate. Agents, cis-2-decenoic acid, D-amino acids, peptides containing D-amino acid residues, gallium mesoporphyrin IX, gallium protoporphyrin IX, curcumin, patuline, penicillic acid, baicalene, naringenin, ursoric acid, aciatic acid, corosolin Agents for dispersing and / or destroying biofilms, mucolytic agents and / or mucilage activators, and / or agents for reducing biofilm formation, selected from acids, amino acids, host defense peptides, and antimicrobial peptides. The pharmaceutical composition according to any one of claims 1 to 25 for use in combination with. Sprayed 4-7% hypertonic physiological saline, metaperiodide, sodium dodecyl sulfate, sodium hydrogencarbonate, tromethamine, silver nanoparticles, bismuth-thiol, ethylenediamine tetraacetic acid, gentamicin-supported phosphatidylcholine-modified gold nanoparticles, chelate. Agents, cis-2-decenoic acid, D-amino acids, peptides containing D-amino acid residues, gallium mesoporphyrin IX, gallium protoporphyrin IX, curcumin, patuline, penicillic acid, baicalene, naringenin, ursoric acid, aciatic acid, corosolin Agents for dispersing and / or destroying biofilms, mucolytic agents and / or mucilage activators, and / or agents for reducing biofilm formation, selected from acids, amino acids, host defense peptides, and antimicrobial peptides. The combination drug according to claim 26 or 27 for use in combination with. 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. 25. The combination pharmaceutical 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. The infectious disease is caused by a strain belonging to the genus Mycobacterium selected from the nontuberculous mycobacteria and tubercle bacilli (Mycobacterium tuberculosis) group and combinations thereof. The pharmaceutical composition according to claim 30, which is triggered. The infectious disease is caused by a strain belonging to the genus Mycobacterium selected from the nontuberculous mycobacteria and tubercle bacilli (Mycobacterium tuberculosis) group and combinations thereof. The combination drug for use according to claim 31, which is triggered. The nontuberculous mycobacteria include Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium avium, and Mycobacterium avium. (Mycobacterium leprae)), as well as the pharmaceutical composition for use according to claim 32, selected from combinations thereof. The nontuberculous mycobacteria include Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium avium, and Mycobacterium avium. (Mycobacterium leprae)), as well as the combination drug for use according to claim 33, selected from combinations thereof. 32. The infection is an opportunistic infection selected from pulmonary MAC disease and nontuberculous mycobacteriosis in patients with cystic fibrosis, chronic obstructive lung disease, or acquired immunodeficiency syndrome. Pharmaceutical composition for use in. 33. The infection is an opportunistic infection selected from pulmonary MAC disease and nontuberculous mycobacteriosis in patients with cystic fibrosis, chronic obstructive lung disease, or acquired immunodeficiency syndrome. Combination medicine for use. The pharmaceutical composition for use according to claim 36, wherein the infection is an opportunistic infection of nontuberculous mycobacteria in a patient with cystic fibrosis. The combination drug according to claim 37, wherein the infectious disease is an opportunistic infection of nontuberculous mycobacteria in a patient with cystic fibrosis. A system for use in imparting antibacterial activity in the treatment or prevention of lung infections caused by mycobacteria or other Gram-positive bacteria, wherein the system is:
1)
(A) With therapeutically effective doses of clofazimine;
(B) With a nonionic surfactant having a hydrophilic-lipophilic balance value of more than 10;
(C) An aqueous liquid carrier selected from water, isotonic saline, buffered saline, and aqueous electrolyte solution.
With atomized combination medicines, including
2) Atomizer and
Including
The clofazimine is present in the form of a suspension and
Moreover, the aerosol particles produced by the system have an aerodynamic mass center diameter of 1 to 5 μm. The use according to any one of claims 28 to 39, wherein the spraying device has an ejection speed of 0.1 to 1.0 ml / min. The use according to any one of claims 28-39, or 41, wherein the total inhalation volume is between 1 ml and 5 ml. The composition comprises bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazineamide, riffampine, moxifloxacin, levofloxacin, and para-aminosalicylate. The use according to any one of claims 28, 30, 32, 34, 36, or 38, which is administered before, simultaneously, or after administration of the agent selected from these mixtures. The combination includes bedaquiline or a pharmaceutically acceptable salt or derivative thereof, cefoxitin, amikacin, clarithromycin, pyrazineamide, riffampine, moxifloxacin, levofloxacin, and para-aminosalicylate, and these. The use according to any one of claims 29, 31, 33, 35, 37, or 39, which is administered before, simultaneously with, or after administration of the agent selected from the mixture of. The pharmaceutical composition according to any one of claims 1 to 25 for use in the treatment and / or prevention of a pulmonary fungal infection or Clostridium difficile or a combination thereof. 26, 27, 29, 31, 33, 35, 37, or 39 of 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 combinations thereof. Combination drug. The pharmaceutical composition according to claim 45, for use in the treatment and / or prevention of pulmonary fungal infections. The combination pharmaceutical according to claim 46 for use in the treatment and / or prevention of pulmonary fungal infections. The pharmaceutical composition according to claim 45 or 47, wherein the lung fungal infection is Candida albicans or Aspergillus fumigatus or a combination thereof. The combination drug according to claim 46 or 48, wherein the lung fungal infection is Candida albicans or Aspergillus fumigatus or a combination thereof. The use according to claim 43, wherein 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 a mixture thereof. 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. A method for treating or preventing a lung infection in a patient in need thereof, comprising administering the composition according to any one of claims 1 to 25 by inhalation. The infectious disease is caused by a strain belonging to the genus Mycobacterium selected from the nontuberculous mycobacteria and tubercle bacilli (Mycobacterium tuberculosis) group and combinations thereof. The treatment or prevention method according to claim 53, which is triggered. The nontuberculous mycobacteria include Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium avium, and Mycobacterium avium. (Mycobacterium leprae)), and the treatment or prevention method according to claim 54, which is selected from a combination thereof. 35. The infection is an opportunistic infection selected from pulmonary MAC disease and nontuberculous mycobacteriosis in patients with cystic fibrosis, chronic obstructive pulmonary disease, or acquired immunodeficiency syndrome. The described treatment or prevention method. The treatment or prevention method according to claim 56, wherein the infection is an opportunistic infection of nontuberculous mycobacteria in a patient with cystic fibrosis. Select from salts of bedaquiline or pharmaceutically acceptable derivatives thereof, cefoxitin, amikacin, clarithromycin, pyrazineamide, rifampine, moxifloxacin, levofloxacin, and para-aminosalicylate, and mixtures thereof. A mycobacteria or other gram in a patient in need thereof, comprising administering the composition according to any one of claims 1-25 by inhalation before, simultaneously, or after administration of the agent to be administered. How to treat or prevent lung infections caused by positive bacteria. The therapeutic or prophylactic method of claim 58, wherein the agent is bedaquiline or amikacin. The therapeutic or prophylactic method of claim 59, wherein the agent is bedaquiline. The following steps:
(1) A step of homogenizing a suspension of clofazimine, the nonionic surfactant, and water to obtain a suspension containing clofazimine of an appropriate particle size.
(2) The step of adjusting the pH of the suspension obtained from (1) to a pH between pH 5.5 and pH 7.5, and
(3) Steps to adjust the sodium chloride concentration to an appropriate concentration,
(4) Steps to adjust the weight osmolality to an appropriate level,
The process for preparing the pharmaceutical composition according to any one of claims 1 to 13, which comprises. The following steps:
(1) A step of homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension containing clofazimine of an appropriate particle size.
(2) The step of isolating the clofazimine and
(3) The step of adding the clofazimine to the nonionic surfactant and water, and
(4) The step of adjusting the pH of the suspension obtained from (3) to a pH between pH 5.5 and pH 7.5, and
(5) Steps to adjust the sodium chloride concentration to an appropriate concentration,
The process for preparing the pharmaceutical composition according to any one of claims 1 to 13, which comprises. The following steps:
(1) A step of pulverizing clofazimine to obtain clofazimine having an appropriate particle size, and
(2) The step of adding the clofazimine to the nonionic surfactant and water, and
(3) A step of adjusting the pH of the suspension obtained from (2) to a pH between pH 5.5 and pH 7.5, and
(4) Steps to adjust the sodium chloride concentration to an appropriate concentration,
The process for preparing the pharmaceutical composition according to any one of claims 1 to 13, which comprises. Appropriate granules of the suspension containing clofazimine in water containing the nonionic surfactant, an appropriate concentration of sodium chloride and adjusted to a pH between pH 5.5 and pH 7.5. The process for preparing a pharmaceutical composition according to any one of claims 1 to 13, comprising homogenizing to obtain clofazimine of diameter. The process of 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. The process of claim 64, wherein the pH is 7.4 and the suitable concentration of sodium chloride is 154 mM sodium chloride. 33. The process of claim 63, wherein the pulverization of clofazimine is performed by jet mill milling, spray drying, ball mill milling, or supercritical fluid treatment. 13. Process. The process of any one of claims 61, 62, 64, 65, 66, or 68, wherein the homogenization of clofazimine is performed by multi-step homogenization. The process according to claim 63 or 20, wherein the pulverization of clofazimine is carried out by multi-step pulverization. The process according to any one of claims 61 to 70, wherein the clofazimine having an appropriate particle size is a particle having an average diameter of less than 5 μm and a D90 of less than 6 μm. The process according to any one of claims 61 to 70, wherein the clofazimine having an appropriate particle size is a particle having an average diameter of less than 2 μm and a D90 of less than 3 μm. The following steps: (a) homogenizing the suspension of chloride, said nonionic surfactant, and water to obtain a suspension containing chloride of appropriate particle size; (b). A step of adjusting the pH of the obtained suspension to a pH between pH 5.5 and pH 7.5; (c) a step of adjusting the sodium chloride concentration to an appropriate concentration, and (d) a weight osmolal concentration. Steps (b), (c), and (d) include (b), (c), (d); (b), (d), (c), including the step of adjusting to an appropriate level. (C), (b), (d); (c), (d), (b); (d), (b), (c); or (d), (c), (b) The process for preparing the pharmaceutical composition according to any one of claims 1 to 13, which can be carried out in sequence. The following steps: (a) homogenizing a suspension of clofazimine and a non-aqueous liquid to obtain a suspension containing clofazimine of appropriate particle size; (b) isolating the clofazimine. And; (c) the step of adding the clofazimine to the nonionic surfactant and water; (d) the step of adjusting the pH of the obtained suspension to a pH between pH 5.5 and pH 7.5. And; (e) including the step of adjusting the sodium chloride concentration to an appropriate concentration; and; in steps (d) and (e), in the order of (d), (e); or (e), (d). The process for preparing the pharmaceutical composition according to any one of claims 1 to 13, which can be carried out. The following steps: (a) the step of pulverizing clofazimine to obtain clofazimine of an appropriate particle size, and (b) the clofazimine containing the nonionic surfactant, an appropriate concentration of sodium chloride. The process for preparing the pharmaceutical composition according to any one of claims 1 to 13, comprising the step of adding to water, the pH of which is adjusted between pH 5.5 and 7.5.