Methods to reduce pneumonia

Abstract

Methods for treating or preventing inflammation in the lungs, or at least for providing a supplement or alternative to previously known methods of treatment, are provided. The present invention provides a method for treating or preventing inflammation in the lungs by administering high concentrations of an inhalable chelating agent, particularly CaEDTA.

Description

[Technical Field]

[0001] This invention treats or prevents inflammation in the lungs by administering a high concentration of inhaled chelating agents. This invention relates to a method for preventing inflammation in the lungs, and to a formulation used in this method. The condition is associated with or caused by cystic fibrosis. [Background technology]

[0002] Cystic fibrosis is characterized by susceptibility to infections that cause inflammation and lung damage. Furthermore, inflammation and lung damage can also occur in the absence of bacterial infection (Sly et al., Am J). Respir Crit Care Med. 2009, Vol. 180 (No. 2): 146~ 52 pages).

[0003] Inflammation is the body's response to an invasion, which includes infections, trauma, and hypersensitivity. Inflammatory responses are complex and involve various mechanisms for defense against pathogens and tissue repair. (Lung) Inflammation is usually caused by pathogens, or by toxins, contaminants, irritants, and allergens. It is caused by exposure.

[0004] During inflammation, many types of inflammatory cells are activated. Each of these is involved in cytokines and signaling. They release substances that modify the activity of other inflammatory cells. The organization of these cells and molecules is This leads to the progression of inflammation. Clinically, acute inflammation can lead to pneumonia and acute respiratory distress syndrome (ARDS). While it is seen in diseases such as asthma, cystic fibrosis, and chronic obstructive inflammatory bowel disease, chronic inflammation is seen in asthma, cystic fibrosis, and chronic obstructive inflammatory bowel disease. It appears in diseases such as lung disease (COPD). The lungs are important organs for gas exchange, Excessive inflammation can be life-threatening. A delicate balance between inflammation and anti-inflammatory factors is crucial for the lungs. It is essential for homeostasis.

[0005] Immunity includes innate and adaptive systems. Innate immunity is nonspecific and is a response to pathogens. It triggers a rapid response, including inflammation, when faced with an invasion. Adaptive immunity is antigen-specific. This adaptive immunity first detects a specific antigen, and then mobilizes inflammatory cells to address that specific antigen. It targets specific antigens. The innate and adaptive systems share components and are able to withstand pathogens. They work together in a coordinated manner for defense.

[0006] The airway epithelium contains various substances, such as mucin, defensin, lysozyme, lactoferrin. They secrete substances such as nitric oxide, and these substances protect the respiratory system from microbial attack. It provides targeted protection. Epithelial cells also transmit several signaling molecules, such as reactive oxygen radicals. Itokines (TNF-α, IL-1β, granulocyte / macrophage colony-stimulating factor [GM]) -CSF) and other factors, as well as platelet-activating factors, attract inflammatory cells to the site of inflammation. Cytokines stimulate the release of arachidonic acid from membrane lipids, leading to the production of eicosanoids. This further stimulates mucus secretion due to goblet cell and tissue inflammation.

[0007] Surfactants are located on the surface of the alveoli and contain four types of surfactant proteins (SP A-D). They possess. Important for reducing tension on the lung surface is that these proteins form boundaries on the alveolar surface. It plays a crucial role in surfactant absorption. SP-A and SP-D also protect the host. They also participate in the process. These bind to bacterial surface molecules and modulate leukocyte activity, and pathogens It causes opsonization of the body.

[0008] IgA secreted by plasma cells forms an additional epithelial protective barrier, which prevents the attachment of microorganisms to the epithelial surface. IgA also binds to pathogenic bacteria, causing phagocytosis and antibody-dependent cell-mediated cytotoxicity. Immunoglobulin E (IgE) induces immediate-type hypersensitivity in the respiratory organs. IgE produces a vigorous reaction by binding to IgE receptors on the surfaces of mast cells, basophils, eosinophils, and B lymphocytes. Repeated exposure to the same antigen induces degranulation and the release of inflammatory mediators including histamine, prostaglandins,

[0009] leukotrienes, and tryptase. These increase vascular permeability, bronchoconstriction, and inflammatory cell infiltration. The iron chelating agent present in the formulation of US Patent Application Publication No. 2016 / 0263151 functions to provide a synergistic effect when

[0010] combined with acidified nitrite, enhancing the ability of the antibiotic to act. Current treatments for lung inflammation include oral or inhaled steroids and non-steroidal drugs that target the host's inflammatory response. However,

[0011] the action is temporary, requires continuous treatment, and has significant side effects.

[0012] This invention treats inflammation in the lungs by administering a high concentration of inhaled chelating agents. They are seeking to provide improved or alternative methods for prevention.

[0013] Prior considerations of the background technology are intended solely to facilitate understanding of this invention. It is reasonable to assume that any of the materials mentioned are part of common general knowledge as of the priority date of the application. This does not constitute an endorsement or permission of, or having been part of, that something. [Overview of the project]

[0014] This invention treats or prevents inflammation in the lungs by administering a high concentration of inhaled chelating agents. To provide a method to prevent this.

[0015] Preferably, the high concentration of the inhaled chelating agent exceeds 37.5 mg / dose.

[0016] Preferably, the inhaled chelating agent is administered at concentrations exceeding 50 mg / dose.

[0017] In one embodiment of the present invention, the high-concentration chelating agent is administered at a dose of at least 50 mg / dose, or 5 The product is provided in a dosage form containing between 0 mg / dose and 300 mg / dose. The chelating agent should be administered up to approximately 1,200 mg / day, preferably at least 150 mg / day of total The dose may be administered between 1 and 4 times daily.

[0018] In one embodiment of the present invention, the high-concentration chelating agent is at least 37.5 mg / dose or The formulation is provided containing between 37.5 mg / dose and 300 mg / dose. The chelating agent Up to approximately 1,200 mg / day, preferably at least 150 mg / day of the total dose taken daily. It may be administered over four doses.

[0019] Preferably, a chelating agent is administered at a dose of 37.5 mg / day to 1,200 mg / day. Alternatively, at least 50 mg / day of a chelating agent is administered. The chelating agent is approximately 1,20 The total daily dose up to 0 mg / day can be administered between 1 and 4 times daily.

[0020] Preferably, each dose of the chelating agent is administered over a period of 8 hours or less. Chelating agents and / or antibiotics are administered over a period of time of one hour or less.

[0021] Preferably, the chelating agent is CaEDTA.

[0022] This invention treats inflammation in the lungs by administering a high concentration of inhaled chelating agents. A method of prevention in which the treatment or prevention of inflammation results in an increase in forced expiratory capacity (FEV1). Further methods will be provided.

[0023] The present invention also treats inflammation in the lungs by administering a high concentration of inhaled chelating agents. or a method of prevention, where the treatment or prevention of inflammation involves matrix metalloproteinases ( This invention provides a method that involves reducing MMP activity.

[0024] This invention treats inflammation in the lungs by administering a high concentration of inhaled chelating agents. A preventive method in which the treatment or prevention of inflammation involves a reduction in the production of hydroxyl radicals. Further methods will be provided.

[0025] The present invention provides an inhalation formulation containing a high concentration of a chelating agent.

[0026] This invention provides an inhalation formulation containing a high concentration of a chelating agent, and provides a high concentration of a chelating agent for inhalation. The drug can be delivered as a single dose.

[0027] The present invention relates to (i) an inhalation formulation containing a high concentration of a chelating agent, and (ii) for use A kit is provided, equipped with instructions, for treating or preventing inflammation in the lungs.

[0028] The present invention relates to (i) an inhalation that can deliver a high concentration of chelating agent as a single dose. (ii) instructions for use, comprising (ii) a required preparation, for treating or preventing inflammation in the lungs We will provide a kit.

[0029] High-concentration chelating agents in the manufacture of inhaled formulations for treating or preventing inflammation in the lungs Use.

[0030] To treat or prevent inflammation in the lungs, a high concentration of inhaled chelating agents is administered as a single dose. Use of inhaled chelating agents in the manufacture of pharmaceuticals for delivery.

[0031] Further features of the present invention are described in the following description of some non-limiting embodiments. It is fully described. This description is included solely for the purpose of illustrating the present invention. This description does not include the broad overview, disclosure, or description of the present invention presented above. It should not be understood as something restrictive. See the attached diagram for an explanation. [Brief explanation of the drawing]

[0032] [Figure 1]This figure shows that submicron particles of EDTA kill Pseudomonas aeruginosa biofilms and act synergistically with tobramycin in vitro. Pseudomonas aeruginosa biofilms in CF mucus were treated with aerosolized EDTA particles and / or tobramycin. The final concentration of tobramycin in droplets was 325 μg / ml. Figure 1A) Confocal microscopy image of the biofilm stained with BacLight LIVE / DEAD; Figure 1B) Bacterial count showing the quantitative effect of the treatment. [Figure 2] This figure shows that CaEDTA reduces the amount of bacteria in CF lungs more quickly than antibiotic treatment alone. Patients with CF were treated with spray-type CaEDTA (EDTA) or saline solution (placebo), and the amount of bacteria in sputum mucus was monitored (colony-forming units per gram of mucus). [Figure 3] Figure 3A shows the mean change in FEV1 (percentage points) over the entire 10-week period (4 weeks post-treatment) in patients treated with CaEDTA or placebo, starting from the beginning of treatment. Figure 3B shows the relationship between improvement in FEV1 (0-2 weeks) and body weight. [Figure 4] This figure shows the EDTA concentrations achieved in the sputum of three CF subjects 5 minutes and 2 hours after treatment with 75 mg of sprayable CaEDTA. [Figure 5] This study demonstrates that administration of high concentrations of chelating agents to mouse lungs reduces inflammation in the absence of infection. The total white blood cell counts are shown for bronchoalveolar lavage fluid (BALF) obtained from mice exposed to air or cigarette smoke (CS) and treated with intranasal administration of either vehicle or deferoxamine (DFO). As expected, cigarette smoke induces an increase in white blood cell counts, but DFO treatment significantly reduces this effect. [Figure 6]We have demonstrated that administration of high concentrations of chelating agents to the lungs of mice reduces inflammation in the absence of infection. The lung weights of mice exposed to air or CS and treated with vehicle or DFO as described above are shown. Since increased weight indicates more inflammation, lung weight can be used as a surrogate for inflammation. In CS-treated mice, lung weight increased significantly, as expected, while DFO treatment reduced average weight, suggesting reduced inflammation. [Figure 7] This study demonstrates that administering high concentrations of chelating agents to mouse lungs reduces inflammation in the absence of infection. CS increases BALF iron content, while treatment with deferoxamine reduces this effect. Left: Average iron content for each mouse group; Right: Scatter plot of the same data. [Modes for carrying out the invention]

[0033] [Methods of treatment or prevention] This invention treats inflammation in the lungs by administering a high concentration of inhaled chelating agents. ) or provide a method to prevent it.

[0034] Preferably, the high concentration of the inhaled chelating agent exceeds 37.5 mg / dose.

[0035] Preferably, the inhaled chelating agent is administered at concentrations exceeding 50 mg / dose.

[0036] It has been previously shown that inhaled EDTA alone does not treat bacterial infections (Bro wn et al. (Am J Dis Child, 1985, Vol. 139 (No. 8): pp. 836-839) );Hassett (Front Microbiol. 2016, vol. 7: p. 291) Brown et al. (1985) used a spray to treat children with chronic Pseudomonas aeruginosa infections. Sodium EDTA was used for 3 months, and no changes in lung function were observed. Concentration-dependent bronchoconstriction (Beasley et al. (Br Med J (Clin Res E)) d) In 1987, Volume 294 (No. 6581): pp. 1197-1198)) causing E Other studies have reported that DTA has no effect on FEV1 (Asmus et al., (J Allergy Clin Immunol. 2001, Vol. 107 (No. 1): 68~ (Page 72)) Therefore, chelating agents can affect, for example, cystic fibrosis (CF). asthma, chronic obstructive pulmonary disease (COPD), or other lung conditions that cause or are associated with inflammation. There is no basis for believing that it has any positive effect on things that are in that state. However, surprisingly, the present invention allows the inhaled chelating agent to treat or prevent pneumonia. They discovered that...

[0037] It is commonly believed that the lung environment of CF is acidic. However, CF lungs are the same as normal lungs. It has recently been shown that it has H (Schultz et al., "Airway sur face liquid pH in children with cystic fi "brosis," Nature Communications, 2017, Vol. 8 (Issue 1) (p. 1409). Therefore, current technologies using acidified nitrite, for example, US patents What is discussed in Patent Publication No. 2016 / 0263151, for example, is that the formulation is acid The conditioned state does not persist, but immediately returns to the normal lung pH of 7.4, thus in CF It is unlikely to work as a floor.

[0038] Although the acidity of the lungs in CF patients was normal, it was found that the iron levels were significantly different from those of normal lungs. Stites et al. (Am J Respir Crit Care Med. 19) In 1999, Volume 160 (Issue 3): pp. 796-798, the iron level in the lungs of CF patients was... Furthermore, it was shown that the levels of iron in the lungs of smokers were significantly higher compared to healthy individuals. The majority of the ferrous phosphate is in the form of ferrous iron, Fe(II), and it has been shown to correlate significantly with disease severity. (Hunter et al., MBio. 2013, Vol. 4 (No. 4): pp. 1-8). Daiichi Iron This generates highly reactive oxygen radicals that can severely damage tissues and DNA. It can participate in the Enton reaction (Jomova et al., Toxicology., 20 11, Volume 283 (Issue 2-3): Pages 65-87; MacNee, Eur J Pharm acol., 2001, Volume 429 (Nos. 1-3): Pages 195-207).

[0039] Without being constrained by theory, the method of the present invention (i) by chelation of zinc (ii) Inactivation of matrix metalloproteinases (MMPs), (ii) Chelation of iron Reduction of reactive oxygen species (ROS) production, and / or (iii) iron and zinc, etc. By reducing the amount of bacteria in the lungs by depriving bacteria of key ions, inflammation is reduced. It is thought that the action within individual lungs is one of the theorized ways in which inflammation is reduced. It may be any combination of these.

[0040] Inhalation is a localized method of administration, and therefore, it does not reach the target area, namely the lungs. It may be more effective in reaching its target and provides high and localized concentrations of inhaled chelating agents. Inhalation can be used to avoid undesirable side effects from systemic exposure to the active substance and to help the patient. This reduces the risk of developing resistance.

[0041] This invention treats or prevents inflammation in the lungs by administering a high concentration of inhaled chelating agents. A method of prevention, further providing a way in which the treatment or prevention of inflammation leads to an increase in FEV. do.

[0042] This invention treats or prevents inflammation in the lungs by administering a high concentration of inhaled chelating agents. A method of prevention is provided, further offering a method in which the treatment or prevention of inflammation is accompanied by a decrease in MMP activity. It is said that matrix metalloproteinases (MMPs) cause lung damage (Garr att et al., Eur Respir J. 2015, Vol. 46 (No. 2): pp. 384-384) and MMP activity is Zn 2+ Dependence on (Hazra et al., Molecular Vi sion, 2012; Vol. 18: pp. 1701-1711) is publicly known. However, in the lungs Previous attempts to target MMPs have been unsuccessful. This invention chelates zinc in the lungs. Inhaled chelating agents are used to reduce MMP-induced lung injury and treat or prevent inflammation. To prevent.

[0043] This invention treats or prevents inflammation in the lungs by administering a high concentration of inhaled chelating agents. A method of prevention in which the treatment or prevention of inflammation is accompanied by a reduction in the production of hydroxyl radicals. Further methods are provided. Since Fe catalyzes the formation of hydroxyl radicals, iron is used in lung damage. The main factors in (Stites et al., (Am J Respir Crit Ca re Med., 1999, Vol. 160 (No. 3): pp. 796-798. However, antioxidants The clinical trial has so far not shown any significant improvement in lung function. Akira used an inhaled chelating agent to chelate the iron in his lungs, and thus hydroxyl To reduce dikal-induced lung injury and treat or prevent inflammation.

[0044] This invention treats or treats infections in the lungs by administering a high concentration of inhaled chelating agents. A method of prevention, in which the treatment or prevention of inflammation is carried out by the presence of a chelating agent, and bacteria in the lungs Further provides methods resulting from the removal or reduction of biofilms generated by these processes. To reduce biofilms, bacteria and biofilms are removed through coughing and sputum. This will enable an increase.

[0045] This invention treats or prevents inflammation in the lungs by administering a high concentration of inhaled chelating agents. A method of prevention, where the treatment or prevention of inflammation stimulates local inflammation and causes local tissue damage. This can cause the removal of protease enzymes produced by bacteria, which can neutralize antibiotic activity. This further provides methods brought about by reduction. These enzymes are primarily cation-dependent. It is expected that removing cations from the environment will deactivate these enzymes. It can be done.

[0046] Preferably, the chelating agent is an iron chelating agent or a zinc chelating agent. More preferably Chelating agents are chelating agents for both iron and zinc (iron / zinc chelating agents). In addition, chelating agents are mixtures of two or more chelating agents, for example, iron chelating agents. A chelating agent and a zinc chelating agent, or an iron / zinc chelating agent and a zinc chelating agent, or an iron chelating agent It may be a mixture of an iron / zinc chelating agent.

[0047] The chelating agent is preferably citric acid, phosphate, or ethylenediaminetetraacetic acid (EDTA). Disodium salt, trisodium salt and tetrasodium salt of ) and calcium of EDTA N,N,N',N' salt, ethylene glycol-bis-(b-aminoethyl ether)-N,N,N',N' -Tetraacetic acid (EGTA); 1,2-bis(2-aminophenoxy)ethane-N,N,N', N'-tetraacetic acid (BAPTA); ethylene-N,N'-diglycine (EDDA); 2,2' -(ethylenediimino)-dibutanoic acid (EBDA); Lauroyl EDTA; Dilauroyl EDTA, Triethylenetetramine dihydrochloride (dihydrochioride)(TRIEN), diethylenetriaminepentaacetic acid (D PTA, triethylenetetramine hexaacetic acid (TTG), deferoxamine (DFO), de Ferrasirox (DSX), dimercaprol, zinc citrate, penicillamine (peni Cilamine, succimer, editronate, sodium hexametaphosphate, ede Calcium disodium toate, D-penicillamine, polyphenols, gallol, catechol Lu, dimercaprol, tetrathiomolybdate, lactoferrin, and cryoquinol Selected from the group consisting of these and combinations thereof.

[0048] Preferably, the chelating agent is a pharmaceutically acceptable chelating agent.

[0049] In one embodiment, the chelating agent is ethylenediaminetetraacetic acid (EDTA). Another embodiment In its form, the chelating agent is deferoxamine (DFO). In another embodiment, The drug is deferasirox (DSX).

[0050] Preferably, the chelating agent has approximately the same iron affinity as EDTA, and / or EDTA It has approximately the same zinc affinity. Formation constant for EDTA at 25°C and 0.1M. Alternatively, the stability constant (log K1) is Fe 2+ For Fe, it is 14.3. 3+ to The ratio for the other substance is 25.1, and for zinc it is 16.5.

[0051] In one embodiment, the chelating agent is a calcium salt of a chelating agent. Preferably, The drug is CaEDTA.

[0052] In one embodiment, the chelating agent is administered at a dose of 37.5 mg / dose to 300 mg / dose for 50 minutes. g / dose to 300 mg / dose, approximately 75 mg / dose to 200 mg / dose, approximately 75 mg Between g / dose and 100 mg / dose, approximately 37.5 mg / dose and 200 mg / dose, approximately 5 Between 0 mg / dose and 200 mg / dose; preferably about 37.5 mg / dose, 50 mg / dose Includes doses of 75 mg / dose, 100 mg / dose, 200 mg / dose, or 300 mg / dose. It is provided in an inhalation dosage form. The chelating agent contains at least 37.5 mg / dose. Preferably provided in an inhalation dosage form. The chelating agent is at least 50 mg / dose. Preferably, it is provided in an inhalation dosage form containing the agent.

[0053] The total amount of chelating agent inhaled per day is preferably about 37.5 mg / day to 120 mg / day. During the 0mg / day period, approximately 50mg / day to 1,200mg / day, approximately 100mg / day to 1,0 Between 00mg / day, approximately 300mg / day to 900mg / day, approximately 400mg / day to 800mg / day Between mg / day; preferably about 150 mg / day, 300 mg / day, 500 mg / day, or 6 mg / day. The dosage is 00 mg / day.

[0054] The total amount of medication inhaled per day is preferably about 0.1 mg of chelating agent / kg (body weight). ) Between 15 mg chelating agent / kg (body weight), approximately 0.5 mg chelating agent / kg (body weight) Between doses of 10 mg chelating agent / kg (body weight), approximately 1.0 mg chelating agent / kg (body weight) to 5 mg g chelating agent / kg (body weight); approximately 1.0 mg chelating agent / kg (body weight) to 3.5 mg Between chelating agent / kg (body weight); preferably about 1.0 mg chelating agent / kg (body weight), 1 0.5 mg chelating agent / kg (body weight), 2.0 mg chelating agent / kg (body weight), 2.5 mg Chelating agent / kg (body weight), 3.0 mg chelating agent / kg (body weight), 3.5 mg chelating agent Chelating agent / kg (body weight), 4.0 mg chelating agent / kg (body weight), 4.5 mg chelating agent / kg (body weight), 5.0 mg chelating agent / kg (body weight), 10 mg chelating agent / kg (body weight), The dosage is 15 mg of chelating agent per kg (body weight).

[0055] If a 75mg chelating agent, such as CaEDTA, is inhaled, the concentration will be approximately 0.4mM. It has been determined that a 1.34 mM chelating agent can be detected in sputum from the lungs after 5 minutes.

[0056] Inhaled chelating agents are administered at intervals of 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, and 1 hour. Preferably over a period of time of 45 minutes, 30 minutes, 20 minutes, 15 minutes, 10 minutes, or 5 minutes or less. It is delivered. When administration is by delivery of dry powder, the inhaled chelating agent is delivered for a period of several seconds. For example, one puff of an aerosol device or dry powder inhaler takes 1 second. It can be delivered over time, with one or more puffs administered at each time point.

[0057] Preferably, the inhaled chelating agent is administered for at least 28 consecutive days. Delivery will take 2 days or more, 3 days, 4 days, 5 days, 6 days, or 7 days. It is possible to inhale chelating agents for 2 to 28 days, or for 1 week, 2 weeks, 3 weeks or It can be delivered in 4 weeks.

[0058] Some subjects require higher doses or more frequent administration over a period of several days or weeks. In subsequent lower doses or maintenance doses, antibiotics and / or chelating agents are administered to the target. You can benefit from the "loading" period.

[0059] Therefore, the present invention is • Deliver a total dose of inhaled chelating agent between 37.5 mg / day and 1,200 mg / day. • Administered at least once a day and up to six times a day, preferably up to four times a day. ru, • Administered over a period of 8 hours or less.

[0060] Preferably, the present invention is • Deliver a total dose of inhaled chelating agent between 37.5 mg / day and 1,200 mg / day. • Administered once or twice per day. • Each dose is administered over a period of less than one hour. • Contains CaEDTA as a chelating agent.

[0061] The preferred amount of any chelating agent is such that the chelating capacity of the drug is compared to that of CaEDTA. It can then be calculated by comparing it with the number given above and multiplying it by the dose range. The result is approximately equal to the preferred level of chelation provided by a preferred amount of EDTA. It should provide a high level of chelation.

[0062] Preferably, the infections are cystic fibrosis (CF); asthma; and chronic obstructive pulmonary disease (COPD). Pulmonary hypertension; lung cancer; pulmonary fibrosis; bronchiectasis; acute respiratory distress syndrome; tuberculosis; non-tuberculous migraine Cobacterial (NTM) lung infections; but not limited to these, including ventilator-associated pneumonia and community-acquired infections. Pneumonia including infectious pneumonia, bronchopneumonia, and lobar pneumonia; bacteria, such as species of the genus Pseudomonas. Streptococcus pneumoniae, Chlamydia, Mycoplasma Zuma pneumonia (Mycoplasma pneumonia), species of Staphylococcus, species of Klebsiella (la spp), Escherichia coli (E. coli), Stenotrophomonas genus (Stenotrophomonas spp), and Aspergillus, Scedosporium, and Candida Infections caused by fungi, including species of the genus Candida (sp); infections that occur, for example, during intubation or Preventive measures or prevention for conditions that may occur in patients on mechanical ventilation; lung transplant patients Infectious diseases in adults; bronchitis; pertussis (whooping cough); inner ear infections; streptococcal throat infections; lung carcinoma It is caused by or associated with lung conditions such as carbuncle; tularemia; or sinusitis. To cause or be associated with these.

[0063] Preferably, the formulation is administered to the target person in need, between approximately 1 to 6 times per day. More preferably, it is administered about four times a day.

[0064] Instead, the formulation is delivered to the target recipient via continuous inhalation or nebulizer. It can be administered. The sprayed formulation can be administered for 24 hours, 12 hours, 8 hours, 6 hours, and 4 hours. It can be delivered in 2 hours or 1 hour, and each of these deliveries is (24 and 12 (Apart from the time limit) it can be repeated several times within a 24-hour period.

[0065] The target group is typically chelating agents at approximately 0.01–15 mg / kg / day, with a tolerance of ±20% or ±10%. You will receive the administration of [the drug]. This administration will be by spraying or using an aerosol device. It is usually administered by at least one, preferably several, "puffs." For example, This involves a single dose of 0.1 mg / kg chelating agent to 15 mg / kg over a day, or Multiple doses can be administered.

[0066] The total daily dose is preferably administered at least once per day, but per day The dose may be divided into two or more doses. Some subjects may require a period of several days or several weeks. Higher doses or more frequent administrations in between, followed by lower doses or maintenance doses. The subject can benefit from the duration of "loading" the chelating agent. Since conditions like vascular disease and COPD are usually chronic, the target group is those who have been receiving such treatment for a long period of time. It is expected to be subject to legal action.

[0067] Regardless of the form of the drug formulation, the particle size is approximately 0.1 μm to 12 μm, or approximately 0.25 μm to 6 μm. Inhalation droplets, preferably in the range of 1 μm to 6 μm, more preferably in the range of about 2 μm to 4 μm. It is preferable to produce particles. Alternatively, the particles are 0.1 μm to 1.0 μm, 0.2 μm m~0.9μm, 0.3μm~0.8μm, 0.4μm~0.7μm, or 0.5μm This may be the case. By creating inhalable particles with a relatively narrow range of sizes, This will further increase the efficiency of drug delivery systems and improve the reproducibility of medication administration. It is possible. Therefore, the particles are 0.1 μm to 12 μm or 2 μm to 6 μm or approximately 3 to 4 μm. In addition to having a size in the range of m, the average particle size is within a narrow range, which is the target 80% or more of the particles delivered are within ±20% of the average particle size, preferably the average It is preferable that the particle diameter be within ±10%, more preferably ±5%, of the particle size. .

[0068] "Particle size" is a concept introduced to compare the dimensions of solid particles and liquid particles (droplets). For droplets and aerosols, the "aerodynamic diameter" and "aerodynamic mass center" are used. Terms such as "particle size diameter (MMAD)" are used. The definitions are given below.

[0069] "Aerodynamic diameter" is the diameter of a unit-density sphere having the same critical sedimentation velocity as the particle in question. Yes, it exists. It is used to predict where such particles will accumulate in the respiratory system.

[0070] The "aerodynamic median particle diameter" is the geometric mean aerodynamic diameter. (50% by weight) The particles in the 50% weight are smaller than those in the MMAD, while the particles in the 50% weight are larger.

[0071] During particle size measurement experiments, the suspension contains countless particles of different sizes that are in motion. When a particle size analyzer analyzes these particles, the particle size analyzer forms a particle distribution curve, The curve starts from the smallest particles, which can be as small as 1 nm, and goes up to the largest particles, which can be as small as 100 μm. It also encompasses the entire particle size range, including large particles. In the particle size distribution curve, the cumulative frequency is shown relative to the particle size. D 10 This means that 10% of the particles in the suspension are smaller than this specific particle diameter. This refers to a specific particle diameter, or a particle having an equal diameter.

[0072] D 50 :D 10 Similarly, D 50 This is the cutoff for 50% of the particle population in the formulation. is the diameter such that 50% of the particles in the suspension have a diameter smaller than or equal to this specific particle diameter refers to this specific particle diameter.

[0073] D 90 :D 90 is the cut-off diameter for 90% of the particle population in the formulation, and 90% of the particles in the suspension have a diameter smaller than or equal to this specific particle diameter refers to this specific particle diameter.

[0074] The term "respiratory organ" is taken to mean the system of cells and organs that function in respiration. In particular, the organs, tissues, and cells of the respiratory system include the lungs, nose, nasal passages, paranasal sinuses, upper pharynx, larynx, trachea, bronchi, bronchioles, respiratory bronchioles, alveolar ducts, alveolar sacs, alveoli, pulmonary alveoli (type 1 and type 2), ciliated mucous epithelium, mucous epithelium, squamous epithelial cells, mast cells, goblet cells, and intraepithelial dendritic cells.

[0075] In one form of the present invention, a method of treating or preventing inflammation in the lungs of a subject comprises administering an inhalable chelating agent at a therapeutically effective or prophylactically effective concentration in a form of one or more administrations of at least 37.5 mg / dose, and the administration or each administration of the chelating agent is administered over a period of 8 hours or less.

[0076] In one form of the present invention, a method of treating inflammation in the lungs of a subject comprises administering an inhalable chelating agent at a therapeutically effective concentration in a form of one or more administrations of at least 37.5 mg / dose and the administration or each administration of the chelating agent is administered over a period of 8 hours or less.

[0077] In one embodiment of the present invention, an inhaled chelating agent at a prophylactically effective concentration is administered, at least 37 Administering 0.5 mg / dose once or multiple times will treat the inflammation in the lungs. A method to prevent the disease, which involves administering a chelating agent or administering each agent within a period of 8 hours or less. A method of administration.

[0078] In one embodiment of the present invention, a method for treating or preventing inflammation in the lungs of a target is such This includes treating or preventing inflammation in the lungs of a person requiring treatment.

[0079] The term "therapeutic effective dose" as used herein, according to the desired dosing regimen. If administered, it will at least partially achieve the desired therapeutic effect or the onset of inflammation. To delay, or hinder progress, or to partially or completely stop the start or progress. This means a sufficient amount of the preparation.

[0080] The term "prophylactic effective dose" as used herein, according to the desired dosing regimen. If administered, it can at least partially prevent inflammation or delay its onset. This means a sufficient amount of the preparation.

[0081] As used herein, “to treat” or “to treat” means to inhibit a disease or condition. To stop the onset of the disease or at least one of its clinical or asymptomatic symptoms. It refers to reducing or lessening a disease or condition. "To treat" or "treatment" means to lessen the disease or condition. To reduce, that is, to reduce the disease or condition or its clinical or asymptomatic symptoms. It further refers to causing another regression. Benefits for the subject being treated. This is statistically significant for the subjects and / or physicians, or at least perceptible. It is possible. In the context of treating inflammation, the term treatment refers to leukocyte infiltration (macrophages). Includes polymorphonuclear cells, neutrophils, lymphocytes, and other immune cells; immunoglobulins; induce inflammation Proactive cytokines and chemokines and their receptors; noxious signaling molecules, for example, ROS and proteolytic enzymes, etc.; abundance and activity of MMPs; oxidative stress Carr; and reducing or eliminating one or more of bronchial hypersensitivity and exacerbations. This includes removing. The term "treatment" refers to increasing anti-inflammatory cytokines and lung function (F This further includes one or more of the increases in EV1).

[0082] Based on the above, treating a single subject using multiple different treatment and administration methods Those skilled in the art will understand that this is possible. Therefore, such drugs, for example, static Subjects who have already received intravenous ciprofloxacin or antibiotics, etc., should use the present invention. Some individuals may benefit from inhaling the agent. The serotonin can be administered via inhalation. This is suitable for patients with cystic fibrosis. They may have the following symptoms, or have been diagnosed with a lung infection, or have high concentration A medical condition in which a patient can benefit from the administration of a chelating agent to the target patient. The formulation of the present invention may also be used for diagnostic purposes. In one embodiment, For example, a subject receives administration of the formulation of the present invention as part of a procedure to diagnose a lung infection. This can result in improvement of one or more of the target symptoms in response to the formulation.

[0083] [formulation] The present invention provides an inhalation formulation containing a high concentration of a chelating agent.

[0084] The inhalation formulation may be in the form of a dry powder for inhalation or in the form of an inhalation spray. Preferably The formulation is indicated for inhalation to treat or prevent inflammation in the lungs.

[0085] In one embodiment, the chelating agent is a calcium salt of a chelating agent. Preferably, The drug is CaEDTA.

[0086] Preferably, the high concentration of the inhaled chelating agent exceeds 37.5 mg / dose. High-concentration inhaled chelating agents exceed 50 mg / dose. Preferably, high-concentration chelating agents This is between 37.5 mg / dose and 300 mg / dose, and between 50 mg / dose and 300 mg / dose. Between approximately 75 mg / dose and 200 mg / dose, between approximately 75 mg / dose and 100 mg / dose. Between approximately 50 mg / dose and 200 mg / dose; preferably approximately 50 mg / dose and 75 mg Preparations containing / dose, 100 mg / dose, 200 mg / dose, or 300 mg / dose The chelating agent is provided in an inhalation dosage form containing at least 37.5 mg / dose. Preferably provided: an inhalation dose containing at least 50 mg / dose of the chelating agent. Preferably provided in this form.

[0087] The total amount of chelating agent inhaled per day is preferably about 37.5 mg / day to 1,200 mg During the period of mg / day, 50 mg / day and 1,200 mg / day, approximately 100 mg / day to 1,000 mg Between mg / day, approximately 300 mg / day to 900 mg / day, approximately 400 mg / day to 800 mg / day For a period of time of day; preferably about 300 mg / day, 500 mg / day, or 600 mg / day. The chelating agent should be used up to approximately 1,200 mg / day, preferably at least 150 mg / day for the entire duration of use. It can be administered in large quantities.

[0088] The total amount of chelating agent inhaled per day is preferably about 37.5 mg / day to 120 mg / day. During the 0mg / day period, approximately 50mg / day to 1,200mg / day, approximately 100mg / day to 1,0 Between 00mg / day, approximately 300mg / day to 900mg / day, approximately 400mg / day to 800mg / day Between mg / day; preferably about 150 mg / day, 300 mg / day, 500 mg / day, or 6 mg / day. The dosage is 00 mg / day.

[0089] The total amount of chelating agent inhaled per day is preferably about 0.1 mg of chelating agent / kg. Between (body weight) and 15 mg chelating agent / kg (body weight), approximately 0.5 mg chelating agent / kg (body weight) Between 10 mg chelating agent / kg (body weight) and approximately 1.0 mg chelating agent / kg (body weight). Between ~5 mg chelating agent / kg (body weight); approximately 1.0 mg chelating agent / kg (body weight) ~3. Between 5 mg chelating agent / kg (body weight); preferably about 1.0 mg chelating agent / kg (body weight) ), 1.5 mg chelating agent / kg (body weight), 2.0 mg chelating agent / kg (body weight), 2. 5 mg chelating agent / kg (body weight), 3.0 mg chelating agent / kg (body weight), 3.5 mg chelating agent Chelating agent / kg (body weight), 4.0 mg chelating agent / kg (body weight), 4.5 mg chelating agent / kg (body weight), 5.0 mg chelating agent / kg (body weight), 10 mg chelating agent / kg (body weight) The dosage is 15 mg of chelating agent per kg (body weight).

[0090] For example, a 50 mg dose of CaEDTA is administered as a 4 mL 33 mM spray solution. (molecular mass C) 10 H 12CaN2Na2O8 has a concentration of 274.27 g / mol. Similarly, a 75 mg dose may be administered as 4 ml at 50 mM, or 100 mg The drug may be administered as 4 ml at a concentration of 66 mM.

[0091] Preferably, the formulation is administered to the target person in need, between approximately 1 to 6 times per day. More preferably, it is administered about four times a day.

[0092] Instead, the formulation is delivered to the target of need via continuous inhalation, via a nebulizer. It can be administered. The spray formulation can be administered for 24 hours, 12 hours, preferably 8 hours, or 6 hours. It can be delivered in 4 hours, 2 hours, or 1 hour, and each of these deliveries can be (24 hours) This can be repeated several times within a 24-hour period (separate from the 12-hour period).

[0093] The formulations of the present invention are disposable packages and portable, handheld, battery-powered devices, for example. If so, it can be administered to the target using an AERx device or the like (U.S. Article 5,820). Patent No. 3,178, Aradigm, Hayward, Calif.). Instead, the present invention Formulation can also be done using mechanical (non-electronic) devices. Conventional jet-type nebulizers... Lizers, ultrasonic nebulizers, soft mist inhalers, dry powder inhalers (DPIs) , including metered-dose inhalers (MDIs), condensed aerosol generators, and other systems The formulation can be delivered using an inhalation device.

[0094] For use as an aerosol, the compounds of the present invention are pressurized in a solution or suspension. In the aerosol container, along with the conventional adjuvant, an appropriate spray agent, such as propane, It can be packaged together with hydrocarbon sprays such as butane or isobutane. Lypowder inhalers are pressurized to produce dry powder particles of pharmaceutical formulations in extremely small volumes. It is a system that can operate using an air supply source. For inhalation, the system is a single-dose system. Multiple chambers, each having a selective element for releasing a pharmaceutical formulation and a single dose. Or it has a blister.

[0095] Aerosols can be produced by forcing a drug through the pores of a membrane, and these pores It has a size in the range of approximately 0.25 to 6 μm (US Patent No. 5,823,178). If the pores are of this size, the particles that escape through the pores and create the aerosol are 0 The particles will have a diameter in the range of 0.5 to 12 μm. Drug particles will have particles within this size range. It can be released with an airflow intended to maintain that state. The generation of small particles is approximately 800 to approximately This is facilitated by the use of vibration devices that provide vibration frequencies in the range of 4000 kilohertz. This can be done. Some adjustments are made to parameters, such as the size of the pores through which the drug is released, and vibration. This is done in terms of dynamic frequency, pressure, and other parameters based on the density and viscosity of the formulation. Those skilled in the art will recognize that it is possible to do so, but the objective of some embodiments is about 0. The objective is to provide aerosolized particles having a diameter in the range of 5 to 12 μm. Please be mindful of this.

[0096] [Excipients] The above-described examples of formulations described herein are well known to those skilled in the art of pharmaceutical science. It can be manufactured by the method described herein. Furthermore, the formulations described herein are as described herein. The listed formulations may contain other optional excipients that are useful in the manufacture and / or administration of the formulations. Yes, it is possible. Non-limiting examples of such excipients are well known in the art, and in the manufacture of formulations and / Or, flavoring agents, coloring agents, parathants, antioxidants, viscosity modifiers, isotonic agents, which are useful for administration. Drug carriers, sustained-release agents, comfort enhancers, emulsifiers, solubilizers, lubricants, binders and other Contains stabilizers.

[0097] Preferably, the formulation of the present invention is sterile. In another embodiment, the formulation of the present invention is stable. Yes, they are.

[0098] Furthermore, the pH level of the formulation can be adjusted by adding a buffer. Preferably, The formulation of the invention is tris(hydroxymethyl)aminomethane (TRIS, which is THAM) It is also known to contain tromethamine as a buffer. TRIS is made with EDTA It may have a further effect of increasing the bacterial killing effect. Preferably, TRIS is used in the formulation. Buffering, the use of EDTA and / or antibiotics in the treatment or prevention of bacterial infections It is added to the formulation of the present invention for both to increase efficacy and for other purposes.

[0099] Furthermore, the formulation of the present invention may contain an antimicrobial preservative.

[0100] Preferably, the pH of the formulation of the present invention is between about 6.5 and 8.0, more preferably about 7.0. The pH is between ~7.4. Bacteria become more resistant to antimicrobial therapy as the pH decreases. This has been previously determined. The preferred pH is for high concentrations of inhaled nitrite in the absence of acidified nitrite. The chelating agent helps to avoid bacterial resistance in formulations containing antibiotics in combination. Keru.

[0101] In one alternative embodiment, the formulation of the present invention is a preservative, a suspending agent, a wetting agent, an isotonic agent and / or may contain diluents. The formulations provided herein are intended for use when administered by inhalation. Physically acceptable, approximately 0.01% to approximately 90%, or approximately 0.01% to approximately 50%, or Approximately 0.01% to 25%, or approximately 0.01% to 10%, or approximately 0.01% to 5% It may contain one or more pharmacologically appropriate suspension fluids. Pharmacologically suitable fluids for use are not limited to these, but include hydroxyl groups or others. The solvent includes, but is not limited to, compounds containing polar groups, and polar solvents. , but not limited to, water or alcohol, such as ethanol, isopropanol , as well as propylene glycol, polyethylene glycol, polypropylene glycol Glycol ethers, glycerol and polyoxyethylene alcohol This includes Cole, etc. Polar solvents are also, but are not limited to, water, one or more Aqueous physiological saline, alcohol, glycol, or pharmaceutically acceptable salt(s)(s) This includes a protic solvent containing a mixture of these. In one alternative embodiment, the present invention Water for use in formulations is subject to applicable legal requirements for use in inhaled drugs. The term must satisfy or exceed the specified value.

[0102] In one embodiment, the formulation described herein is aqueous and contains 0-90% water. It may have. In other embodiments, the aqueous formulations described herein contain 20-80% It may contain water. In yet another embodiment, the aqueous formulation may contain 50-70% water. The water is freshwater, distilled, sterilized, demineralized, or deionized water. It could also include...

[0103] Instead, the formulation is non-aqueous, containing no water or only a very small amount of water. It may be included (e.g., less than 1%, less than 0.1%, less than 0.01%).

[0104] In one embodiment, the formulation comprises one or more pharmaceutically or physiologically acceptable carriers. The body further comprises a diluent or excipient.

[0105] In addition to or instead of sterilization, the formulation of the present invention minimizes the possibility of microbial contamination. It may contain pharmaceutically acceptable preservatives. Furthermore, it may contain pharmaceutically acceptable The stability of the formulation can be increased by using a preservative in the formulation of the present invention. However, Since the treated tissue may be sensitive to irritants, storage may be optional for safety reasons regarding inhalation. It should be noted that a suitable preservative must be selected. The appropriate preservative for use in this specification is: This includes, but is not limited to, those that protect the solution from contamination with pathogen particles, and phenyl Ethyl alcohol, benzalkonium chloride or benzoic acid, or benzoate, e.g. For example, it includes sodium benzoate and phenylethyl alcohol. In embodiments, the formulations of this specification contain approximately 0.001% to approximately 10.0% w / w benzalkonium chloride. It contains conium, or about 0.01% v / w of phenylethyl alcohol. The preservative is also , approximately 0.001% to approximately 1%, preferably approximately 0.002% to approximately 0.02%, more preferably It can be present in an amount of 0.02% w / w.

[0106] The formulations provided herein also range from approximately 0.001% to approximately 90%, or approximately 0.00 1% to approximately 50%, or approximately 0.001% to approximately 25%, or approximately 0.001% to approximately 10% Alternatively, it may contain one or more types of emulsifiers, wetting agents, or suspending agents in an amount of approximately 0.001% to 1%. This is also acceptable. Such drugs for use in this specification are not limited to these, but Polyoxyethylene sorbitan fatty acid esters or polysorbates, for example, not limited to these. Although not specified, polyethylene monooleate sorbitan (polysorbate 80), poly Rubate 20 (Polyoxyethylene (20) monolaurate sorbitan), Polysorbate T65 (Polyoxyethylene (20) Sorbitan Tristearate), Polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (20) sorbitan monopal Mitart, containing sorbitan polyoxyethylene (20) monostearate; Reci Chin; agar; carrageenan; locust bean gum; guar gum; tragacanth; acacia Xanthan gum; Indo-rubber; Pectin; Amidized pectin; Ammonium phosphate Acid; microcrystalline cellulose; methylcellulose; hydroxypropylcellulose; hydr Roxypropyl methylcellulose; ethyl methylcellulose; carboxymethylcellulose Sodium, potassium, and calcium salts of fatty acids; monoglycerides and Diglycerides; Acetates of fatty acid monoglycerides and diglycerides; Mo Lactic acid esters of noglycerides and diglycerides; monoglycerides and diglycerides of fatty acids Citrate esters of cerides; tartaric acid esters of fatty acid monoglycerides and diglycerides. Monoacetyl tartrate esters and diglycerides of fatty acids Acetyl tartrate esters; a mixture of monoglycerides and diglycerides of fatty acids. tartaric acid esters; sucrose esters of fatty acids; sucrose glycerides; polyglycerides of fatty acids Lycerol esters; polyglycerol esters of castor oil polyconcentrated fatty acids; fatty acids Propane-1,2-diol ester; sodium stearoyl-21 acrylate (a ctylate; calcium stearoyl-2-lactylate; stearoyl tartrate ;Sorbitan monostearate;Sorbitan tristearate;Sorbitan monolaurate n; sorbitan monooleate; sorbitan monopalmitate; quillaja extract; soybean oil Polyglycerol esters of dimerized fatty acids; oxidatively polymerized soybean oil; and pectin extract Includes discounted items.

[0107] The formulation of the present invention inhibits mucosal drying and prevents irritation by containing approximately 0.001% by weight. May contain 5% by weight of a humectant: sorbitol, propylene glycol, polyethylene glycol. Various pharmaceutically acceptable humectants, including cholang, glycerol, or mixtures thereof. The discrepancy can also be utilized.

[0108] The formulations of the present invention contain a bronchodilator, another anti-inflammatory agent, a surfactant, aspirin, or ethinopropyl alcohol. It may further contain adjuvants such as alcohol.

[0109] The bronchodilators used in the formulations of the present invention are not limited to these, but may be used in some cases. , β2-adrenergic receptor agonists (e.g., albuterol, vanbuterol, s) Rubutamol, salmeterol, formoterol, alformoterol, levosalbutamol Procaterol, indacaterol, carmoterol, milbeterol, procaterol (e.g., lorol, terbutaline), and antimuscarinic agents (e.g., throspium, ipratro). (Contains pium, glycopyrronium, acridinium, etc.) Use of drug combinations It is possible.

[0110] Additional anti-inflammatory agents that may optionally be used in the formulations of the present invention are not limited to those mentioned above. However, inhaled corticosteroids (e.g., beclomethasone, budesonide, ciclesonide, Fluticasone, etipredonol, mometasone, etc., leukotriene receptor antagonists Stromal and leukotriene synthesis inhibitors (e.g., montelukast, dileuton, ibuzil) (Sto, Zafirlukast, Pranlukast, Amervant, Typercast, etc.), Sic oxygenase inhibitors (e.g., ibuprofen, ketoprofen, ketrolac, i Ndomethacin, naproxen, zaltoprofen, lornoxicam, meloxicam, cele Coxib, Lumiracoxib, Etricoxib, Piroxicam, Ampiroxicam, Synth Xicam, diclofenac, felbinac, lornoxicam, mesalazine, triflusar This includes drugs such as thinolysine, iguratimod, and pamicogrel. This can be done. Aspirin, which acts as an anti-inflammatory agent, can also be added.

[0111] The surfactants included in the present invention are not limited to these, but also include synthetic surfactants (E xosurf (registered trademark), dipalmitoylphosphatidylcholine and oleic acid Includes. Drug combinations can be used.

[0112] Antioxidants, such as glutathione and vitamin E, zinc and zinc salts of EDTA. It can be added.

[0113] Ethyl alcohol vapor acts as an antifoaming agent in the lungs, making phlegm more liquefied, and this This can assist breathing and reduce pulmonary edema. Ethanol is added to the formulation of the present invention. Between 0.5% and 60%, more preferably between 1% and 40%, between 1% and 20%, or between 1% and 1% It can be added to bring the concentration to between 0%. Ethanol can be added to 1%, 2%, 3%, 4%, 5%. %, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40 It can be added to make a percentage of 45%, 50%, 55%, or 60%.

[0114] The present invention also relates to high concentrations of chelating agents in combination with other drugs administered by inhalation. Regarding use: These other drugs require a suitable delivery vector, e.g., plasmid or virus. It may contain nucleotide sequences that can be incorporated into other drugs such as ruth vectors. It reduces the viscoelasticity of mucus and uses therapeutic nucleotide sequences (DNA, RNA, siRNA). Enzymes such as DNase and other mucolytic agents upregulate chloride ion channels. Chemicals that increase the flow of ions across cells, such as nicotine, P2Y2A Gonists, for example, elastase inhibitors containing α-1 antitrypsin (AAT), N- Acetylcysteine, antibiotics and cationic peptides, e.g., lanthobiotics, Specifically, duramycin, short-acting bronchodilators (e.g., β2-adrenaline) Receptor agonist-like albuterol or indacaterol), M3 muscarinic antagonist Nist (e.g., ipratropium bromide), K + - Channel opener, long-acting Bronchodilators (e.g., formoterol, salmeterol), steroids (e.g., bude) (e.g., Sonide, fluticasone, triamcinolone, beclomethasone, ciclesonide), Xa Thrutin, leukotriene antagonists (e.g., montelukast sodium), phospho Diesterase-4 inhibitors, adenosine receptor antagonists, and various other anti-inflammatory drugs (for example) For example, a Syk kinase inhibitor (AVE-0950), a tryptase inhibitor (AVE-892) 3 and AVE-5638), tachykinin antagonist (AVE-5883), inducible Nitric oxide synthase inhibitors (GW-274150 and others), transcription factor decoys, TLR-9 agonists, antisense oligonucleotides, siRNA, DNA, CGR P, lidocaine, reverse β2 agonist, anti-infective oxidative therapy, cytokine modulator (For example, CCR3 receptor antagonists (GSK-766994, DPC-168, A ZD-3778), TNF-α production inhibitors (LMP-160 and YS-TH2), (IL-4 antagonist (AVE-0309)), IgE small molecule inhibitor, cell adhesion Molecular (CAM) inhibitors, small molecule targeting VLA4 receptor or integrin inhibitors. Fa.4.β.1 (e.g., R-411, PS-460644, DW-908e and C Includes DP-323, which blocks T cell signaling by inhibiting calcineurin. Immunomodulatory substance (tacrolimus), heparin neutralizer (talactoferrin α), cytosol The PLA2 inhibitor (Efipladib) or a combination thereof may be required. If the subject has CF, the subject may also be combined with the formulation of the present invention in accordance with standard practice. In addition, standard drugs such as ibakhtol, pulmozyme, mannitol, or others Approved drugs can also be administered.

[0115] The delivery of combination products involves combining drugs into a single stable formulation or delivering them at the time of administration. Provide the drugs in separate containers to be combined, or instead sequentially produce the products This can be achieved by delivery.

[0116] Preferably, the formulation of the present invention is stable. When used herein, the following applies: The stability of the formulation being offered refers to the stability of the active pharmaceutical ingredient, such as chelating agents and antibiotics, at a given temperature. This refers to the length of time that more than 80%, 85%, 90%, or 95% of the initial amount is present in the formulation. For example, the formulations provided herein can be stored at temperatures between approximately 15°C and 30°C. Often, it remains stable for at least 1, 2, 12, 18, 24, or 36 months. Furthermore, the formulation must be used after storage at 25°C for more than 1, 2, 12, 18, 24, or 36 months. It may be suitable for administration to the target. In another alternative embodiment, Arrhen Using IUS kinetics, 1, 2, 12, 18, 24 or between approximately 15°C and approximately 30°C Even after storage of the formulation for more than 36 months, the initial amount of the active pharmaceutical ingredient (e.g., chelating agents and antibiotics) remains 8 More than 0%, or more than 85%, or more than 90%, or more than 95% remains.

[0117] When used herein, the statement that a formulation is stable during “long-term storage” means that the formulation The predicted shelf life is that of a usage time of 1, 2, or 3 months at 25°C and If the product has a storage period of 1, 2, or 3 years or more, or equal to that, at 5°C, the product is... This means that it is suitable for administration to subjects that require it. In certain embodiments of this specification So, using Arrhenius kinetics, after such storage, >80% or >85 It is predicted that 0% or >90% or >95% of the chelating agent and the antibiotic remains .

[0118] As used herein, the term "inflammation" refers to one or more signs of the body's response to an assault, such as an infection, an environmental attack (including tobacco smoke), trauma or allergy. Inflammation can be either acute or chronic, and the signs include tissue swelling, recruitment of different types of inflammatory cells, release of cytokines and mediators, and include bronchial allergy. Inflammation can be localized, asymptomatic or transient, or it can be more extensive and become chronic. Inflammation can involve both humoral and cellular immune responses and can persist even after the assault that induced it has been removed. Signs of inflammation include, but are not limited to, an increase in the levels of inflammatory cells (e.g., dendritic cells, macrophages, neutrophils, lymphocytes, eosinophils and mast cells), an increase in the levels of pro-inflammatory cytokines (e.g., TNFα, IL-1β, IL-6, IL-8, and IFNγ) and their receptors, excessive proteases including MMP, ROS and other mediators, and markers of inflammation, such as C-reactive protein (CFP) and sputum and serum calprotectin. Short-term inflammation ("acute") can be accompanied by clinical symptoms including increased respiratory rate and dyspnea, wheezing, coughing, and a decrease in FEV1, resulting in airway swelling, changes in lung compliance, airway reactivity and mucus hypersecretion, and if this persists ("chronic"), it can result in structural damage to the airway wall and lung parenchyma in the form of fibrosis, cystic changes and

[0119] [Method for manufacturing a medicament] High-concentration chelating agents in the manufacture of inhaled formulations for treating or preventing inflammation in the lungs Use.

[0120] A high concentration inhaled chelating agent is used as a single dose to treat or prevent inflammation in the lungs. Use of inhaled chelating agents in the manufacture of pharmaceuticals for delivery.

[0121] Preferably, the high-concentration chelating agent is administered at least 37.5 mg / dose, and at least 50 ml. g / dose, or between 50 mg / dose and 300 mg / dose, or 37.5 mg / dose and The formulation is provided containing 300 mg / dose. The chelating agent is approximately 1,200 mg / dose. The total dose, preferably at least 150 mg / day, is administered 1 to 4 times daily until the day of administration. This is possible. Preferably, the chelating agent is CaEDTA.

[0122] [kit] The present invention relates to (i) an inhalation formulation containing a high concentration of a chelating agent, and (ii) for use A kit is provided for treating or preventing inflammation in the lungs, complete with instructions.

[0123] The present invention relates to (i) an inhalation that can deliver a high concentration of chelating agent as a single dose. (ii) a required preparation and instructions for use for treating or preventing inflammation in the lungs We will provide the kit.

[0124] Preferably, the high-concentration chelating agent is administered at least 37.5 mg / dose, and at least 50 ml. g / dose, or between 37.5 mg / dose and 300 mg / dose, or 50 mg / dose and The formulation is provided containing 300 mg / dose. The chelating agent is approximately 1,200 mg / dose. Until the day, preferably up to the total dose of at least 150 mg / day is administered 1 to 4 times daily. Obtain. Preferably, the chelating agent is CaEDTA.

[0125] In one embodiment, the kit of the present invention comprises a therapeutically effective amount of a high-concentration inhalable chelating agent. Contains the agent. In alternative embodiments, the formulation is pre-measured, pre-mixed, and / or Alternatively, it is pre-packaged. Preferably, the inhalation solution is sterile.

[0126] The kit of this invention also includes instructions designed to facilitate user compliance. May include. Instructions, as used herein, refer to any labels, accompanying documents, etc. The instructions may be placed on one or more surfaces of the packaging material, or on a separate sheet. , or any combination thereof, can be provided. For example, in one embodiment, the present invention The kit includes instructions for administering the formulation of the present invention. In one embodiment, the instructions are provided for administering the formulation of the present invention. This indicates that the preparation is suitable for treating lung inflammation. Such instructions also apply to the preparation. Instructions for administration via nebulizer or dry powder inhaler, as well as instructions for administration via nebulizer or dry powder inhaler. It may also include "show".

[0127] Inhaled chelating agents and any further activators may be prescribed by a physician or user as required by the user. It can be individually packaged so that it can be formulated into pharmaceutical products as needed. Pharmaceutical formulations containing required chelating agents and any additional activators can be packaged together. Therefore, a de minimis preparation by a physician or user is required. In any case, The packaging should maintain the chemical, physical, and aesthetically pleasing integration of the active ingredients.

[0128] [General rules] A person skilled in the art will recognize that the present invention described herein may have modifications and variations other than those specifically described. The present invention includes all such modifications and variations. The present invention also includes all steps, features, formulations and compounds mentioned or shown in the specification, individually or in combination, and any and all combinations or any two or more steps or features. A person skilled in the art will recognize that the present invention described herein may have modifications and variations other than those specifically described. The present invention includes all such modifications and variations. The present invention also includes all steps, features, formulations and compounds mentioned or shown in the specification, individually or in combination, and any and all combinations or any two or more steps or features. A person skilled in the art will recognize that the present invention described herein may have modifications and variations other than those specifically described. The present invention includes all such modifications and variations. The present invention also includes all steps, features, formulations and compounds mentioned or shown in the specification, individually or in combination, and any and all combinations or any two or more steps or features. A person skilled in the art will recognize that the present invention described herein may have modifications and variations other than those specifically described. The present invention includes all such modifications and variations. The present invention also includes all steps, features, formulations and compounds mentioned or shown in the specification, individually or in combination, and any and all combinations or any two or more steps or features. A person skilled in the art will recognize that the present invention described herein may have modifications and variations other than those specifically described. The present invention includes all such modifications and variations. The present invention also includes all steps, features, formulations and compounds mentioned or shown in the specification, individually or in combination, and any and all combinations or any two or more steps or features. A person skilled in the art will recognize that the present invention described herein may have modifications and variations other than those specifically described. The present invention includes all such modifications and variations. The present invention also includes all steps, features, formulations and compounds mentioned or shown in the specification, individually or in combination, and any and all combinations or any two or more steps or features.

[0129] Each document, reference, patent application or patent cited in this text is hereby expressly incorporated herein by reference in its entirety, which means that it should be read and considered as part of this text by the reader. The fact that a document, reference, patent application or patent cited in this text is not repeated in this text is for reasons of brevity only. Each document, reference, patent application or patent cited in this text is hereby expressly incorporated herein by reference in its entirety, which means that it should be read and considered as part of this text by the reader. The fact that a document, reference, patent application or patent cited in this text is not repeated in this text is for reasons of brevity only. Each document, reference, patent application or patent cited in this text is hereby expressly incorporated herein by reference in its entirety, which means that it should be read and considered as part of this text by the reader. The fact that a document, reference, patent application or patent cited in this text is not repeated in this text is for reasons of brevity only. Each document, reference, patent application or patent cited in this text is hereby expressly incorporated herein by reference in its entirety, which means that it should be read and considered as part of this text by the reader. The fact that a document, reference, patent application or patent cited in this text is not repeated in this text is for reasons of brevity only. Each document, reference, patent application or patent cited in this text is hereby expressly incorporated herein by reference in its entirety, which means that it should be read and considered as part of this text by the reader. The fact that a document, reference, patent application or patent cited in this text is not repeated in this text is for reasons of brevity only.

[0130] Any manufacturer's instructions, descriptions, product specifications, and product sheets for any product described herein or in any document incorporated herein by reference can be incorporated herein by reference and used in the implementation of the present invention. ​​​​​​​​​​​​​​​​​​The present invention as described herein relates to one or more range values ​​(e.g., size, position). This can include values ​​such as the voltage and magnetic field strength. A range of values ​​includes all values ​​within that range. It is thought that this includes a value that defines the range, and a value that defines the boundary for that range. The range of adjacent values ​​includes values ​​that produce the same or substantially the same result as the immediately adjacent value. Therefore, unless otherwise indicated, the specification and claims are not described. The numerical parameters listed may vary depending on the desired characteristics to be obtained by this invention. It is an approximation. Therefore, "approximately 80%" means "approximately 80%", and also "80%". At the very least, each numerical parameter is calculated based on the number of significant digits and the usual rounding method. It should be interpreted as follows.

[0133] Throughout this specification, unless otherwise required by context, the word "comprise" is used. " or variations, for example, "comprises" or "comprising" It is thought that this means the inclusion of the integer or group of integers being stated, but any other This does not exclude integers or groups of integers. This disclosure and, in particular, the claims and / or within a paragraph, "comprises", "comprised", "includes" Terms such as "comprising" may have meanings that belong to U.S. patent law. These are "includes," "included," and "incl." It can mean "consisting of," and can mean "essentially consisting of." Terms such as "ially of" and "consists essentially of" are This has implications under U.S. patent law, and for example, these may permit elements that are not explicitly enumerated. However, this does not affect the fundamental or novel features of the present invention that are found in the prior art or that are not present in the present invention. It is also important to focus on eliminating elements that contribute to the problem.

[0134] Other definitions of selected terms used herein are provided in the detailed description of the invention. It can be found within and applies throughout. Unless otherwise defined, it is used herein. All other scientific and technical terms used herein are understood by those skilled in the art of the field to which this invention pertains. It has the same meaning as what is commonly understood. The term "activator" is a type of active It may mean a sexual agent, or it may encompass two or more active agents.

[0135] The following examples are intended to more fully describe a method of using the invention described above. Furthermore, the best mode envisioned for carrying out various aspects of the present invention is described. These methods are not intended to limit the true scope of the invention. Rather, please understand that it is presented for illustrative purposes. [Examples]

[0136] Further features of the present invention are described more fully in the following non-limiting examples. These are included solely for the purpose of illustrating the present invention. This should not be understood as a limitation on Ming's extensive explanation.

[0137] [Example 1] For those who wish to treat or prevent lung infections by administering high concentrations of inhaled chelating agents. law Using suspension droplets of cystic fibrosis mucus collected from epithelial cell lines, a realistic in vi Biofilms were grown in the tro model (Haley et al., BMC Micro Biol, 2012, Vol. 12: p. 181). Pseudomonas aeruginosa clinical strains (MIC tobramycin > 2) Cultures at 56 μg / ml were prepared in M63 without a carbon source to mimic nutrient restriction. They were propagated and then transitioned to the late quiescent phase.

[0138] Suspend a few drops of mucus (5 μl) from an inverted IBIDI coverslip, and 10 3 Colony-forming units (CFUs) were inoculated, and then incubated in a humid environment at 35°C for 72 hours. We baited the film to generate a biofilm. Then, we sprayed droplets with tobramycin (2 Either 0 mg / ml, or aerosolized CaEDTA particles (10 mg / ml), or Both were treated for 5 minutes. The control was treated with a 50 / 50 solution of 0.9% physiological saline / water for spraying. After processing, the droplets were incubated for 16 hours, and then BacLight LIVE / Stained with DEAD (1 μl) and fixed with paraformaldehyde vapor for 30 minutes. Confocal microscopy was performed. We used a microscope to visualize the biofilm.

[0139] Figure 1A shows a thick cell with mostly viable cells (green) after treatment with saline spray. and exhibits a strong biofilm. As expected in resistant strains, tobramycin monotherapy is effective. The treatment in Germany has little effect on the survival rate. EDTA alone kills a certain amount (red It causes blood cells. The combination of tobramycin and EDTA is particularly biofil The majority of the cells have been killed. Figure 1B shows a quantitative representation of the microscopic image in Figure 1A. The control biofilm was 1 × 10 8The CFU / ml was EDTA-tobramycin. The treated biofilm was reduced by more than 6 orders of magnitude, and less than 10 2 The level was reduced to CFU / ml.

[0140] Patients with CF aged ≥ 6 years who were hospitalized due to exacerbation of symptoms were randomly selected and administered intravenously. In addition to these standard treatments with antibiotics and tobramycin spray, EDTA or physiological Administer saline solution (placebo). Administer EDTA together with tobramycin in 4 ml 50 ml doses. M CaNa2EDTA, 111mM Tris in 0.9% physiological saline, pH 7.1, for spraying. It was administered as a solution. After random selection, subjects were treated in the hospital for two weeks, during which time patients received daily treatment. Four treatments were administered (300 mg EDTA / day, or 3.3 mg EDTA / kg / day). Then, the patient was discharged and the treatment was continued twice a day for four weeks. The patient was then monitored for another four weeks. We monitored the data and set the total experimental time to 10 weeks.

[0141] Sputum was induced using 3% hypertonic saline for spraying at a rate of 8-10 L / min for ≥5 minutes. Before treatment Samples were collected and processed according to the relevant protocol at weeks 2, 6, and 10, and then stored at -80°C. It was stored. The mucus was separated from the clear sputum, and sputalysin (1 ml per gram of sputum) was added. Mix with ), vortex, incubate for 1 hour, then Skim Milk G The samples were placed in a lycerol storage medium and stored at -80°C.

[0142] The initial screening, followed by the third clinical examination (around 2 weeks later), the fifth clinical examination (6 weeks later), and finally a follow-up examination. During the follow-up investigation (10 weeks), sputum samples were obtained from the subjects' sputum. Mucus was separated from the clear sputum. Then, treat with Sputalysin (1 ml per gram of sputum) and Skim Milk Placed in a glycerol storage medium (1 ml / 100 mg mucus), and vortexed They were mixed and stored at -80°C.

[0143] Thaw the sample on ice and prepare serial dilutions from 10 to 7 times the original concentration, then prepare 20 μL of each dilution. The solution is cultured in three types of agar: McConkey (McC) agar or Blood agar (BA). The samples were placed on each plate. The plates were incubated at 35°C.

[0144] Species of the genus Pseudomonas are transparent or very pale pink on McC agar plates. Defined as lactose-negative colonies. Coarse-shaped colonies have a metallic sheen. Colonies with rough colony edges and a smooth morphology form regular colonies on McC agar. Colonies with a knee-like boundary, slow growth, and a mucous morphology are formed by the massive secretion of bacteria. It was surrounded by alginate.

[0145] The number of coarse colonies, smooth colonies, and mucilaginous colonies was measured over a 24-hour period. Count on both C agar and BA plates, and incubate the plates again for another 24 hours. The samples were then analyzed and counted to confirm each colony morphology. Ronnie was picked up and arranged in strips on a BA plate to obtain a pure culture.

[0146] Perform Gram staining to confirm that the isolate is Gram-negative and consists of rod-shaped cells. , and by rubbing a very small portion of the colony onto an oxidase test strip Further identification was performed by confirming the oxidase-positive state. Rapid development of dark blue This shows an oxidatively positive isolate.

[0147] Species of the genus Pseudomonas were identified by testing for resistance to the C390 antibiotic. The pure phosphate-buffered saline (PBS) was mixed with a McFarland density of 0.5. A suspension of the isolated strain is diffused on a nutrient agar (NA) plate, and antibiotic-impregnated d The disk was placed. After incubation at 35°C overnight, the area around the disk was cleaned. The absence of an inhibitory zone indicated resistance to antibiotics. Each of the confirmed Pseudomonas species present (likely P. aeruginosa) Pick up a single colony of the morphological type and resuspend it in a glycerol / serum storage medium. Stored at -80°C. Figure 2 shows Pseudomonas aeruginosa at 2 and 6 weeks. a) The change in colony count for (McC) is shown compared to the start of treatment. Two weeks after the treatment, the reduction in colony count was 4.5 times compared to the placebo group. The EDTA group showed a ratio of >400 times.

[0148] [Example 2] Treatment of pneumonia leads to a dose-dependent increase in FEV1. Subjects with CF ≥ 6 years of age who were hospitalized due to exacerbation of symptoms were randomly selected and administered intravenously. In addition to these standard treatments with antibiotics and tobramycin spray, EDTA or physiological Administer saline solution (placebo). Administer EDTA together with tobramycin in 4 ml 50 ml doses. A spray solution of M CaEDTA, 111 mM Tris in 0.9% physiological saline, pH 7.1 The drug was administered. After random selection, the subjects were treated in the hospital for two weeks, during which time the patients received the drug four times a day. The treatment was administered (300 mg of EDTA / day). After discharge, the treatment was continued twice a day for 4 weeks. The elephants were monitored for another four weeks, bringing the total experimental time to 10 weeks.

[0149] Lung function was measured by spirometry during the clinical examinations for each experiment. Of the three attempts, the best result was achieved. The data was recorded as is, and the results were expressed as a percentage of the predicted value.

[0150] Figure 3A shows the FEV1 levels for both groups at 2, 6, and 10 weeks after the start of treatment. This shows the average change. The average increase in FEV1 after 2 weeks was 16 points in the EDTA group. In contrast to the placebo group, the difference was 5 percentage points. This difference occurred after the completion of the treatment. The effect lasted for 4 weeks, with a 7 percentage point increase in the EDTA group compared to a 2 percentage point increase in the placebo group. There was an increase in intake. This demonstrates a clear improvement in lung function in the EDTA group. However, only slight changes were demonstrated in the placebo group. Figure 3B shows the EDTA group (R 2 =0 .70) shows an inverse correlation between FEV1 improvement and body weight, but Tobramais This shows that there is no correlation in the placebo group treated with than alone (R 2 (=0.01). This is because EDTA has a dose-dependent effect on lung function (mg EDTA / kg body weight). To indicate that.

[0151] Figure 4 shows that the delivery of 75 mg of CaEDTA to the lungs was 0.41–1.3 minutes after administration. This indicates that it results in a peak EDTA concentration of 4 mM.

[0152] [Example 3] Tobacco smoke-induced pneumonia is treated by administering high doses of chelating agents to the lungs. It is possible The effects of chelating agents on pneumonia were investigated in a mouse model of chronic obstructive pulmonary disease (COPD). It was tested. It is known that tobacco smoke (CS) induces pneumonia, and pneumonia is related to leukemia. This can be measured by an increase in sphere count and lung weight.

[0153] Male BALB / c mice (8 mice per group) were exposed to a specified dose of cigarette smoke (3 sticks) Tobacco, three times a day (Monday to Friday), or exposed to filtered room air for a period of two weeks. The mice were exposed to tobacco smoke 30-60 minutes before each exposure during the experiment. One dose of the iron chelating agent deferoxamine (DFO, 3.8 mg in 50 μl) or vehicle The nasal cavity was then treated. The mice were then sacrificed, and their airways and lungs were examined for tobacco smoke-induced inflammation. The effects on the elements and their concentrations were evaluated.

[0154] Bronchoalveolar lavage fluid (BALF) is collected (approximately 1 ml / mouse), and the lungs are surgically removed. Then, it was weighed. Equal amounts of trypan blue were mixed with BALF and Zeiss Axiosco Using a PE fluorescence microscope, counted manually with a standard Neubauer hemocytometer. The total number of viable cells in BALF was determined by laser ablation-induced coupling. Iron was measured by elemental analysis using plasma mass spectrometry (LA-ICP-MS), and publicly known The metal content was quantified by comparing it to the standard.

[0155] Figure 5 shows, as expected, that cigarette smoke significantly increases the total number of BALF leukocytes. This indicates that treatment with the iron chelating agent DFO significantly reduces this effect.

[0156] In line with this, Figure 6 shows that average lung weight increases significantly with treatment by cigarette smoke. Treatment at CFO has shown that this effect can be prevented.

[0157] As previously described, Stites et al. (Am J Respir Crit Car e Med. 1999, Vol. 160 (No. 3): pp. 796-798) describes the lungs and... The study showed that iron levels in the lungs of smokers are significantly higher compared to healthy individuals. Figure 7 (left) shows that average iron levels in BALF mice exposed to cigarette smoke were significantly reduced. It has been confirmed that the iron content increases, and that treatment with DFO reduces the average BALF iron content. 7 (right) shows that in 6 out of 7 mice treated with DFO (1 mouse was unrelated to the treatment). (Died for no apparent reason), BALF iron content was at the same level as that of mice exposed to air. This indicates that it was a lu.

[0158] [Virtual Example P1: High-dose dry powder chlorine for infection, inflammation, and oxidative stress] [In vivo experiments on the effects of the drug] Subjects with CF requiring treatment with dried tobramycin powder were divided into four cohorts. They will be assigned to receive 112 mg of dried powder twice a day for 28 days. In addition, Cohort 1 For patients (aged 18 and over), the increasing dose of dried CaEDTA powder (37.5 mg BID) is used. Administer 75 mg BID for 2 weeks, then 150 mg BID for 1 week. Patient T2 (>18 years old) received CaEDTA (37.5 mg BID for 1 week; 75 BI) Administer D for 2 weeks; 75 mg QID for 1 week. Cohort 3 (patients aged 12-18 years) This involves CaEDTA (37.5mg BID for 1 week; 75mg BID for 2 weeks, 150 Administer mg BID for 1 week. Finally, the observational cohort will receive tobramycin alone for 2 weeks. Administer for 8 days.

[0159] Sputum samples are collected weekly and evaluated for markers of infection and inflammation. Bacteria are isolated from the sputum colony. - Monitored by count. As a measure of structural damage, matrix metalloprotein The levels of tissue inhibitors of enzymes (MMPs) and metalloproteinases (TIMPs) Using latinzymography and immunoassay, respectively, previously described Measure as described (Gaggar et al., Eur Respir J. 2011, Vol. 38 (3 Issue): pp. 721-727; Garratt et al., Eur Respir J. 2015, Volume 46 (Issue 2): pp. 384-3894). The amount of iron in sputum is as previously described in ICP. -Quantify using MS (Hunter et al., Mbio. 2013, Vol. 4 (No. 4): pp. 1-8) ). Use an immunoassay to assess the amount of iron-binding protein. Myeloperoxida Neutrophil activity is also assayed as a measure of neutrophil inflammation, as previously described (G aggar et al., Eur Respir J. 2011, Vol. 38 (No. 3): 721-72. (Page 7). 3-Chlorotyrosine was measured as a biomarker for hypochlorous acid, a powerful oxidizing agent. Determine the level using stable isotope dilution gas chromatography with mass spectrometry. Measurement (Gaggar et al., Eur Respir J. 2011, Vol. 38 (No. 3): (Pages 721-727). Using a commercially available immunoassay kit, reactive oxygen species (ROS) Protein carbonyls are measured as an indicator (Gaggar et al., Eur Respir J. 2011, Vol. 38 (No. 3): pp. 721-727). Using immunoassays, As previously described, by measuring glutathione (GSSG and GSH) , assess oxidative stress (Kettle et al., Eur Respir J. 2014, Volume 44 (Issue 1): pp. 122-129. Inflammatory and oxidative stress markers (e.g., IL-8) Gene expression of IL-6 and TNFα is also monitored using nanostrings, and protein The quality is measured by ELISA. Oxidative stress can also be transmitted through metabolites, for example, malton Dialdehyde (molondialdehyde) (colorimetric assay) or 8-iso It can be measured using methods such as Plastan (ELISA). Iron is removed by laser-induced coupling. Elemental analysis is performed using plasma-intensity plasma-mass spectrometry (LA-ICP-MS).

[0160] This experiment showed a reduction in inflammatory markers in the EDTA group compared to the placebo group, and A decrease in iron levels is expected. This experiment will compare MMP and TIMP, especially M The change in balance between MP-9 and TIMP-1 (which is associated with the progression of bronchiectasis) Further demonstrations are expected.

[0161] This experiment compared the amount of bacteria in sputum in subjects treated with EDTA compared to control patients. Further reductions and increases in FEV1 are expected.

[0162] [Virtual Example P2: Effects of high-dose chelating agents on inflammation, lung injury, and oxidative stress] [In vitro experiment for use] Lung epithelial cells are grown in tissue culture and exposed to Fe(II) or excess oxygen. Induces inflammation. Cells are treated with CaEDTA (0, 1, 5, 10, 25, 50 mM) for 30 minutes. Process for 1, 3, 24, and 48 hours.

[0163] Immunoassays are used to test for inflammatory markers, such as IL-6, IL-8, and TNF- Monitor alpha, neutrophil elastase, and other changes. Reduced glutathione (GSH) By evaluating apoptosis based on oxidative stress levels and TUNEL assays, To measure treasury and toxicity, in this case both can be measured using commercially available assay kits, for example, The Glutathione Fluores from rmoFisher Scientific cent Detection Kit and BioVision Inc.'s TUNEL We use DNA Gragmentation Assay Kits, etc.

[0164] This experiment showed that inflammatory markers were more concentrated in EDTA-treated cells compared to controls. Substantial reduction; GSH is reduced (this is in EDTA-treated cells compared to the control). This indicates a reduction in reactive oxygen species; and measured by the TUNEL assay. In this case, apoptosis was reduced in EDTA-treated cells compared to the control. It is expected that this will be demonstrated.

[0165] [Virtual Example P3: High-dose chelate for spraying against inflammation, lung injury, and oxidative stress] [In vivo experiments on the effects of the drug] Subjects with CF ≥ 6 years of age who were hospitalized due to exacerbation of symptoms were randomly selected and administered intravenously. In addition to these usual treatments with antibiotics and spray tobramycin, spray EDTA Alternatively, administer physiological saline (placebo). EDTA should be administered together with tobramycin in a dose of 4 ml. 50 mM Ca EDTA, 111 mM Tris in 0.9% physiological saline, pH 7.1, for spraying. Administer as a solution.

[0166] After random selection, subjects will be treated in the hospital for two weeks, during which time they will receive treatment four times a day. (Up to 300 mg EDTA / day, or 3.3 mg EDTA / kg / day). Next, the subjects The patient will be discharged and the treatment will be continued twice a day for four weeks. The subject will be monitored for another four weeks, and the total experimental time will be measured. This should be done for 10 weeks. Induction should be performed with 3% hypertonic saline spray at a rate of 8-10 L / min for ≥ 5 minutes. Sputum is collected by doing so. Samples are collected before treatment and at weeks 2, 6 and 10, related Process according to the protocol and store at -80°C.

[0167] • Expression of inflammatory markers The collected sputum is stored in RNAlater®, and Qiagen RNEas Extract total RNA using y(registered trademark) or a similar extraction kit, and convert it to cDNA. Sivaneson et al. (Mol Microbiol, Vol. 79, pp. 1353-1366) As described in ), qPCR is used to monitor inflammatory markers and known hysteric acid Quantification compared to wool-keeping genes, such as actin and / or GAPDH. do.

[0168] This experiment compared the gene expression of inflammatory markers in the EDTA group compared to the placebo group. This is expected to show an average reduction.

[0169] • Cell damage, free iron, and oxidative stress The collected sputum was frozen without any processing, and as described above, the inflammatory markers were analyzed. As a measure of structural damage, matrix metalloproteinases (MMPs) and The levels of taloproteinase (TIMP) tissue inhibitors were measured using gelatin zymography. The immunoassays are used respectively and measured as previously described (Gagg ar et al., Eur RespirJ. 2011, Vol. 38 (No. 3): pp. 721-727; G arratt et al., Eur Respir J., 2015, Vol. 46 (No. 2): 384~ (Page 94). The amount of iron in the sputum is quantified by ICP-MS as previously described (Hu nter et al., MBio. 2013, Vol. 4 (No. 4): pp. 1-8). Immunoassay used. Then, the amount of iron-binding protein is evaluated. Using an immunoassay, as previously described... As mentioned, oxidative stress can be assessed by measuring glutathione (GSSG and GSH). Evaluate (Kettle et al., Eur Respir J. 2014, Vol. 44 (No. 1): (pp. 122-9).

[0170] This experiment showed a reduction in inflammatory markers in the EDTA group compared to the placebo group. This is expected. This experiment will examine the relationship between MMP and TIMP, particularly between MMP-9 and TIMP-1. It shows a change in the balance between (which is associated with the progression of bronchiectasis). Further predictions are expected.

[0171] [Virtual Example P4: High-dose chelating agent for inflammation, lung injury, and oxidative stress] [In vivo experiments for use] A single-center, randomized, double-blind, crossover study will be conducted targeting cystic fibrosis. Participants were randomized to receive either CaEDTA or saline solution (placebo) for two weeks. Extract. After this, a washout period and then other treatments for 2 weeks (EDTA or (Placebo) continues.

[0172] Monitor iron levels, inflammatory markers, MMP / TIMP, and FEV1 as described above. Yes. As previously mentioned, myeloperoxidase is used as a measure of neutrophil inflammation. Activity is also assayed (Gaggar et al., Eur Respir J. 2011, 3 Volume 8 (Issue 3): pp. 721-727). 3. As a biomarker for the powerful oxidizing agent hypochlorous acid. - Measure chlorotyrosine. Stable isotope dilution gas chromatography using mass spectrometry. - Use this to measure the level (Gaggar et al., Eur Respir J.2011) Year, Vol. 38 (No. 3): pp. 721-727). Using a commercially available immunoassay kit, Protein carbonyls are measured as an indicator of reactive oxygen species (ROS) (Gaggar et al., Eur Respir J. 2011, Vol. 38 (No. 3): pp. 721-727.

[0173] This experiment compared subjects treated with EDTA to a placebo in terms of iron and inflammation levels. Reduction in marker levels, changes in MMP / TIMP balance, and increase in average FEV1. It is expected that this will show an increase. This experiment will reduce myeloperoxidase activity and clo Further reductions in the average levels of rotyrosine and carbonyl are expected.

[0174] Clinical data demonstrate efficacy at 300 mg / day for two weeks. The same experiment... It has been demonstrated that 50 mg / day (75 mg BID) is beneficial for lung function and infections. (Figure 2 shows a decrease in bacterial count; Figure 3A shows an improvement in lung function.) Considering the improvement of a certain magnitude (average FEV1 of 16 percentage points), it will be understood by those skilled in the art. As assumed by the hypothetical example P1, a much lower dose, namely 75 There is a very high probability that mg / day (37.5 mg BID) will be effective.

[0175] Figure 4 shows that 30 minutes after a single dose of 75 mg of CaEDTA, the concentration inside the mucus plug was 1.34 mM. This indicates that EDTA is brought up to this point. The penetration of drugs such as tobramycin into CF sputum is It is significantly delayed (Kuhn, RJ (2001). Formula o f aerosolized therapeutics.Chest Volume 120, 94 (Pages S-98S), therefore, the concentration of EDTA in the airway surface fluid is substantially higher than in the center. It is publicly known that this is the most likely possibility. Therefore, the daily dose is 37.5 mg (clinical merit It is expected to show efficacy in fully intensified experiments (four times lower than the low dose with a positive result). This is reasonable. This is especially true for young patients who are administered at higher doses per unit of body weight. Furthermore, it generally shows a greater therapeutic effect at FEV1 (Figure 3B).

[0176] Without departing from the basic concept of the present invention, based on the above teachings relating to the disclosed invention Many variations and modifications of the modes described above carry out various embodiments of the present invention. The form will be obvious to those skilled in the art. The above embodiments of the present invention are merely illustrative and not limiting. It should never be interpreted as such, and all such modified and modified forms are The properties are considered to fall within the scope of the present invention as determined from the above description.

[0177] The research of this invention relates to Cystic Fibrosis Foundation Ther This project was supported by prize money from apeutics.

Claims

1. An inhalation composition for treating or preventing chronic inflammation of the lungs, comprising a chelating agent ethylenediaminetetraacetic acid (EDTA), wherein the concentration of the chelating agent in the inhalation composition is at least 50 mM, and the inhalation composition is used to deliver the chelating agent at doses of 37.5 mg to 300 mg, and to deliver the chelating agent in each dose over a period of 2 hours or less, wherein each dose of the inhalation composition comprises droplets or particles with a particle size of 0.1 μm to 12 μm.

2. The inhalation composition according to claim 1, wherein the chelating agent is calcium ethylenediaminetetraacetic acid (CaEDTA).

3. The inhalation composition according to claim 1 or 2, wherein the chelating agent is combined with tris(hydroxymethyl)aminomethane (TRIS).

4. The inhalation composition according to any one of claims 1 to 3, wherein the inhalation composition is used for administering the chelating agent once to four times daily.

5. Use of a chelating agent, ethylenediaminetetraacetic acid (EDTA), at a concentration of at least 50 mM, for the manufacture of an inhalation composition for treating or preventing chronic inflammation of the lungs, wherein the inhalation composition is used to deliver the chelating agent at doses of 37.5 mg / dose to 300 mg / dose, the inhalation composition is manufactured to administer the chelating agent in single doses over a period of no more than 2 hours, and the single doses of the inhalation composition each contain droplets or particles with a particle size of 0.1 μm to 12 μm.

6. The use according to claim 5, wherein the chelating agent is calcium ethylenediaminetetraacetic acid (CaEDTA).

7. The use according to claim 5 or 6, wherein the chelating agent is combined with tris(hydroxymethyl)aminomethane (TRIS).

8. The use according to any one of claims 5 to 7, wherein the inhalation composition is used in an application in which the chelating agent is administered once to four times daily.

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