Use of 5-amino-2,3-dihydro-1,4-phthalazinedione in the inhalation treatment of inflammatory lung diseases

Abstract

The present invention relates to the use of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof in the inhalation treatment of inflammatory lung diseases. The present invention particularly relates to the use of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt for this purpose. Advantageous features of an aerosol containing 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof and a method for producing the aerosol are disclosed. The present invention further relates to a kit for the inhalation treatment of inflammatory lung diseases.

Description

[Technical Field]

[0001] The present invention relates to the use of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof in the inhalation treatment of inflammatory lung diseases. The present invention particularly relates to the use of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt for this purpose. Advantageous features of an aerosol containing 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof and a method for producing the aerosol are disclosed. The present invention further relates to a kit for the inhalation treatment of inflammatory lung diseases. [Background technology]

[0002] Pulmonary diseases affect the lower airways of the respiratory system, particularly the lungs. This term encompasses pathological conditions that impair gas exchange in the lungs or bronchi of mammals. Generally, they are distinguished into obstructive and restrictive pulmonary diseases. Obstructive pulmonary diseases are characterized by airway obstruction, which limits the amount of air that can enter the alveoli due to narrowing of the bronchial tree caused by inflammation. Restrictive pulmonary diseases are characterized by a loss of lung compliance, leading to incomplete lung expansion and increased lung stiffness.

[0003] They can also be classified as airway diseases, lung tissue diseases, lung infections, and lung proliferative diseases. Airway diseases affect the tubes that carry oxygen and other gases in and out of the lungs. They usually cause narrowing or obstruction of the airways. Typical airway diseases include asthma, chronic obstructive pulmonary disease (COPD), and bronchiectasis. Lung tissue diseases affect the structure of lung tissue. Tissue scarring or inflammation prevents the lungs from fully expanding, complicating gas exchange. As a result, these patients are unable to breathe deeply. Pulmonary fibrosis and sarcoidosis are typical examples. Lung infections refer to disorders caused by infection of the lower respiratory tract, such as pneumonia. Lung proliferative diseases include all tumors or neoplasms of the lower respiratory tract.

[0004] Most airway diseases are caused by underlying inflammation or at least include an inflammatory component. Lung tissue diseases often also have an inflammatory component, unless caused by direct physical obstruction of the airways. Infectious and proliferative diseases of the lung may also have an inflammatory component, often secondary to infection or underlying malignancy.

[0005] Therefore, these inflammatory lung diseases have in common that they can be pharmacologically treated with anti-inflammatory drugs.However, therapeutic effectiveness is often hindered by the insufficient availability of drugs at the inflammatory site of the lung and their respective efficacy.Systemic administration, such as oral administration or parenteral administration, often does not result in therapeutic success or only insufficient therapeutic success.

[0006] An alternative route of administration is inhalation. Metered dose inhalers (MDIs) are widely used, for example, in the treatment of asthma. They are used to have a container for the pharmaceutical preparation, a canister, a metering valve for measuring the dispensed amount, and a mouthpiece for inhalation. The pharmaceutical preparation consists of a drug, a liquefied gas propellant such as a hydrofluoroalkane, and optionally pharmaceutically acceptable excipients.

[0007] A particular group of MDIs are dry powder inhalers (DPIs). They deliver medication to the lungs in the form of a dry powder. Most DPIs rely on the patient's inhalation force to draw the powder from the device and subsequently break it down into particles small enough to reach the lungs. Therefore, insufficient patient inhalation flow rates can result in reduced dose delivery, incomplete powder breakup, and poor device performance. Therefore, most DPIs require minimal inspiratory effort for proper use. Therefore, their use is limited to older children and adults.

[0008] Disorders affecting the upper part of the bronchi or lower respiratory tract can be addressed in this way, for example, by asthma sprays, but disorders affecting the alveoli where gas exchange takes place can only be treated poorly due to ineffective inhalation administration, e.g., COPD: the administered drug particles, at least in therapeutically effective amounts, are unable to reach the bottom of the lungs by inhalation.

[0009] Nebulizers are used to administer active ingredients in the form of a mist that is inhaled into the lungs. Physically, this mist is an aerosol. It is generated in a nebulizer by breaking down solutions and suspensions into small aerosol droplets that can be inhaled directly through the device's mouthpiece. In conventional nebulizers, the aerosol can be generated by mechanical force, such as spring force in soft mist nebulizers, or electrical force. In jet nebulizers, a compressor actively flows oxygen or compressed air through an aqueous solution at high velocity, thus generating the aerosol. A variant is the pressurized metered dose inhaler (pMDIs). Ultrasonic nebulizers actively use an electronic oscillator that vibrates a piezoelectric element at high frequency to generate ultrasonic waves in a liquid reservoir.

[0010] The most promising technology is the vibrating mesh nebulizer. It uses a mesh polymer membrane with numerous laser-drilled holes. This membrane is placed between a liquid reservoir and an aerosol chamber. A piezoelectric element placed on the membrane induces high-frequency vibrations of the membrane, forming droplets in an aqueous solution and pressurizing these droplets through the membrane's holes into the aerosol chamber. This technology can produce very small droplet sizes. Furthermore, this significantly reduces the patient's inhalation time, a feature that significantly improves patient compliance. Only these mesh nebulizers are believed to be capable of producing droplets with the desired size range containing the active ingredient and delivering them to the patient's alveoli in therapeutically effective amounts within a reasonable time.

[0011] A summary of the characterization of vibrating mesh aerosol generators is provided in Kuo et al. (2019) Aerosol and Air Quality Research 19:1678-1687. Neither the materials of the invention nor specific ranges of aerosol parameters for the materials of the invention are disclosed therein.

[0012] However, not all drugs that may be effective in the pharmaceutical treatment of inflammatory lung diseases are suitable for mesh spraying technology.For example, some drugs are almost insoluble in water, or their inherent physicochemical molecular properties do not allow the production of aerosols with desired particle size range.Therefore, many nebulized anti-inflammatory drugs have only achieved limited success in the treatment of inflammatory lung diseases.

[0013] Therefore, there is a medical need to find pharmaceutical agents that are highly effective in treating inflammatory lung diseases while at the same time allowing for the generation of aerosols in the desired particle size range so that they can reach the alveoli of patients in need thereof.

[0014] Surprisingly, this problem could be solved by nebulization of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts or solvates, hydrates, crystalline polymorphs, tautomers or isotopically enriched forms. DETAILED DESCRIPTION OF THE INVENTION

[0015] 5-Amino-2,3-dihydro-1,4-phthalazinedione belongs to the pharmaceutical class of phthalazinedione. This class of compounds is known for its beneficial anti-inflammatory properties. 5-Amino-2,3-dihydro-1,4-phthalazinedione is also known as luminol. Luminol has excellent chemiluminescent properties. It is widely used in diagnostic assays and forensics as a detection tool, for example, for tracing blood spots. In medicine, 5-amino-2,3-dihydro-1,4-phthalazinedione has been developed in the form of its sodium salt. In several countries, 5-amino-2,3-dihydro-1,4-phthalazinedione, for example (ao), is approved for a wide range of acute and chronic inflammatory disorders, including acute infections of bacterial and viral origin, especially of the intestinal tract, hepatitis B and C, gastroenteritis, prostatitis, endometriosis, inflammation of the throat, bronchial asthma, pneumonia, periodontitis, pyelonephritis, and autoimmune diseases such as Crohn's disease, ulcerative colitis, lupus erythematosus, and scleroderma. Furthermore, the scientific and patent literature continues to contain a long list of therapeutic indications for which 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt has allegedly been tested or has been suggested to be of beneficial use (see, e.g., (ao), WO 2004 / 041169; WO 2007 / 018546; WO 2012 / 127441; WO 2017 / 202496; WO 2018 / 082814).

[0016] For example, systemic administration of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt has been suggested for the treatment of chronic obstructive pulmonary disease, although evidence of its therapeutic efficacy remains lacking.

[0017] Russian Patent No. 2266119C2 describes the use of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium or potassium salts or mixtures thereof in the inhalation treatment of tuberculosis. However, the document does not mention the inhalation method and device by which the substance was administered, nor the composition and parameters of the aerosol used for this purpose.

[0018] However, 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt has not yet been described for use in the prevention or treatment of inflammatory lung diseases by inhalation administration of a nebulized aerosol.

[0019] While most conventional immunomodulatory drugs have serious adverse reactions or are at least problematic with long-term treatment, 5-amino-2,3-dihydro-1,4-phthalazinedione and its pharmaceutically acceptable salts are well tolerated and have a high margin of safety relative to the doses administered.

[0020] To ensure better solubility and bioavailability, pharmaceutically acceptable salts of 5-amino-2,3-dihydro-1,4-phthalazinedione are used. Sodium, potassium, and lithium salts have been described for therapeutic use (see WO 2010 / 082858). The crystal structures of lithium, sodium, potassium, rubidium, and cesium salts are described in Guzei et al. (2013) Journal of Coordination Chemistry 66, 3722-3739. Therefore, this patent application also refers to the use of all pharmaceutically acceptable salts of 5-amino-2,3-dihydro-1,4-phthalazinedione.

[0021] 5-amino-2,3-dihydro-1,4-phthalazinedione is often used as a hydrate, for example, the sodium salt dihydrate. Therefore, this patent application also refers to the use of all hydrates and other solvates of 5-amino-2,3-dihydro-1,4-phthalazinedione and its pharmaceutically acceptable salts. 5-amino-2,3-dihydro-1,4-phthalazinedione and its pharmaceutically acceptable salts can form complexes with suitable ligands. Therefore, this patent application also refers to such complexes.

[0022] Anhydrous formulations are often preferred to ensure reproducible, standardized API manufacturing and provide improved stability characteristics of the active agent. The anhydrous forms of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt are described as crystalline polymorphs in WO 2011 / 107295 (Form I, Form II) and WO 2016 / 096143 (Form III). These polymorphs are substantially free of phase impurities and have been characterized by X-ray powder diffraction. This method yields a set of characteristic d-values, which indicate the interplanar spacing (Å) at which Bragg reflections occur and the corresponding 2-theta (2θ) angles (°). This provides a unique and unambiguous fingerprint for each polymorph.

[0023] For Form I, the following values ​​were determined: d-value: 13.5; 6.9; 5.2; 4.6; 3.9; 3.5; 3.4; 3.3; 3.1; 3.0 and / or 2θ values: 6.5; 12.7; 16.9; 19.3; 22.8; 25.8; 26.6; 27.2; 28.7; 30.3.

[0024] Form II is characterized by the following values: d-value: 12.9; 7.9; 7.1; 6.5; 5.3; 4.0; 3.7; 3.6; 3.3; 3.2 and / or 2θ values: 6.8; 11.2; 12.5; 13.7; 16.7; 22.4; 24.3; 24.9; 27.2; 27.8.

[0025] Form III gave the following values: d values: 13.131; 7.987; 7.186; 6.566; 6.512; 5.372; 3.994; 3.662; 3.406; 3.288; 3.283; 3.222; 3.215; 3.127; 2.889 and / or 2θ values: 6.73; 11.07; 12.31; 13.48; 13.59; 16.49; 22.24; 24.29; 26.14; 27.10; 27.14; 27.67; 27.72; 28.52; 30.93.

[0026] 5-amino-2,3-dihydro-1,4-phthalazinedione itself also exhibits polymorphism: Form I (Paradies (1992) Ber. Bunsen-Ges. Phys. Chem 96:1027-1031) and Form II (WO 2017 / 140430) have been disclosed.

[0027] Therefore, the present patent application also refers to the use according to the invention of all crystalline forms of 5-amino-2,3-dihydro-1,4-phthalazinedione and its pharmaceutically acceptable salts and polymorphs thereof.

[0028] Similar therapeutic effects are known for various phthalazinediones, which are derivatives of 5-amino-2,3-dihydro-1,4-phthalazinedione, and their pharmaceutically acceptable salts. One example is 6-amino-2,3-dihydrophthalazine-1,4-dione (isoluminol). A summary of suitable phthalazinediones is given in WO 2007 / 018546. It is reasonable to assume that these compounds will exhibit similar effects when used in the therapeutic applications according to the present invention.

[0029] Tautomerism refers to the rapid internal transformation of organic compounds in which a hydrogen atom or proton formally migrates to the interior of the compound. This involves switching a single bond and an adjacent double bond. A single form is called a tautomer. For example, keto-enol tautomerism occurs in 5-amino-2,3-dihydro-1,4-phthalazinedione (Proescher and Moody (1939) J Lab Clin Med, 1183-1189). Therefore, this patent application also refers to the use of all tautomers of 5-amino-2,3-dihydro-1,4-phthalazinedione and its pharmaceutically acceptable salts.

[0030] Isomers are a general term for molecules with the same chemical formula but different chemical structures. They can be distinguished into constitutional (structural) isomers (atom or functional group exchanges) and stereoisomers. Stereoisomers can be further divided into enantiomers (non-superimposable mirror images of the same molecule) and diastereomers (the same molecule with different configurations at one or more stereocenters). Diastereomers can be further divided into cis / trans isomers (referring to the relative orientation of functional groups within a molecule), as well as conformational isomers (formally rotations around a single bond) and rotamers (different rotational configurations around a single bond). An example of a constitutional isomer of 5-amino-2,3-dihydro-1,4-phthalazinedione is 6-amino-2,3-dihydrophthalazine-1,4-dione (isoluminol). Stereoisomers of phthalazinedione derivatives can exist. Therefore, this patent application also refers to the use of all isomers of 5-amino-2,3-dihydro-1,4-phthalazinedione, its derivatives, and pharmaceutically acceptable salts.

[0031] In some applications, it may be desirable to use isotopically enriched forms of the compounds of the invention, for example for diagnostic purposes, and therefore the present patent application also refers to such isotopically enriched forms of the compounds of the invention.

[0032] From a pharmacokinetic point of view or for manufacturing rationale, it may be preferable to use a prodrug as a dosage form. A prodrug is administered in a pharmacologically inactive form and is converted in the body into a metabolically active form. This conversion can occur systemically or locally. Therefore, this patent application also refers to prodrugs of the compounds of the present invention.

[0033] As used throughout this application, the term "5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof" is intended to encompass all the aforementioned molecular variants of 5-amino-2,3-dihydro-1,4-phthalazinedione, i.e., 5-amino-2,3-dihydro-1,4-phthalazinedione, or one of its pharmaceutically acceptable salts, or solvates, hydrates, crystalline polymorphs, tautomers or isotopically enriched forms.

[0034] In the scope of this application, an aerosol is a mixture of air and solid or liquid particles. In particular, the term "aerosol containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts" refers to an aerosol produced by nebulization of an aqueous solution containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts.

[0035] Unless otherwise defined, technical or scientific terms used herein have the meanings ascribed to them by one of ordinary skill in the relevant art.

[0036] According to this application, the terms "drug substance," "active substance," "active agent," "pharmaceutically active agent," "active ingredient," or "active pharmaceutical ingredient" (API), when not otherwise specified or used in their generic sense, refer to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof.

[0037] The term "composition" or "pharmaceutical composition" includes at least one active ingredient in any pharmacologically acceptable defined dosage and dosage form together with at least one pharmaceutically acceptable excipient, and also includes any agent produced directly or indirectly from the components outlined below, as a combination, accumulation, complex or crystal, or as a result of other reaction or interaction, as well as optionally at least one additional pharmaceutical drug listed below.

[0038] The term "excipient" is used herein to describe any component of a pharmaceutical composition other than the pharmaceutically active principle. The selection of an appropriate excipient depends on various factors, such as the dosage form, dosage, desired solubility and stability of the composition, etc.

[0039] The terms "effect," "therapeutic effect," "action," "therapeutic action," "efficacy," and "effectiveness," with respect to a substance of the invention or any other active substance referred to herein, refer to a beneficial result that occurs causally in an organism to which the substance has previously been administered.

[0040] According to the present invention, the terms "effective amount" and "therapeutically effective amount" refer to an amount of a substance of the present invention that is large enough to cause the desired beneficial effect in a subject in need of such treatment.

[0041] The terms "treatment" and "therapy" include the administration of at least a substance of the present invention alone or in combination with at least one additional pharmaceutical agent, regardless of the chronological order of administration. Such administration is intended to substantially improve the disease course of inflammatory lung diseases by completely curing the disease or by halting or slowing the progression of damage during the disease course.

[0042] The terms "prevention" or "prophylactic treatment" include the administration of at least a substance of the present invention, alone or in combination with at least one further pharmaceutical agent, regardless of the chronological order of administration, to prevent or inhibit the onset of symptoms resulting from inflammatory lung diseases. It particularly refers to a condition in a patient where the onset of such symptoms is expected with a reasonable probability to occur in the distant or near future.

[0043] The terms "subject" and "patient" include individuals suffering from a disease condition or disorder associated with inflammatory lung disease, whether a confirmed or suspected diagnosis. The individual is a mammal, particularly a human.

[0044] For the purposes of this application, the term "medicine" is intended to include human and veterinary medicine.

[0045] In the context of this patent application, the term "inflammatory disease" or "inflammatory lung disease" refers to a disease, disorder, or other physical condition that manifests itself primarily in pulmonary inflammation. Inflammation is the response of bodily tissues to a stimulus (exogenous or endogenous pathogen) or injury. It can be triggered by physical, chemical, and biological stimuli, including, inter alia, mechanical trauma, radiation damage, caustic chemicals, extreme heat or cold, and infectious agents such as bacteria, viruses, fungi, and other pathogenic microorganisms or parts thereof. Inflammation can have beneficial (e.g., within the scope of wound healing) and / or harmful effects in the affected tissue(s). In the first stage, inflammation is considered acute. If it does not resolve after a period of time, inflammation can become chronic. Typical signs of inflammation are redness, swelling, fever, pain, and decreased function. This can even lead to loss of function in the affected tissue.

[0046] Inflammation is one of the first responses of the immune system, for example, when activated by infected or degenerated endogenous cells. The innate immune system mediates nonspecific responses, particularly general inflammatory responses, while the adaptive immune system provides specific responses to each pathogen, which are then memorized by the immune system. An organism may be in an immunodeficient state, i.e., the immune response cannot satisfactorily cope with the aforementioned stimuli or damage. On the other hand, the immune system may become overactive, altering the defense against endogenous tissues, as in the case of autoimmune diseases.

[0047] In the context of this patent application, the term "degenerative disease" or "degenerative lung disease" refers to a disease, disorder, or other physical condition in which a continuous process leads to degenerative cellular changes. The affected tissue or organ continuously deteriorates over time. Such degeneration can be due to physical or physiological overexertion of certain vulnerable body structures, lifestyle, dietary habits, age, congenital diseases, or other endogenous causes. The degeneration can be caused by or accompanied by atrophy or dystrophy of the respective tissue or organ, especially the lung. In many cases, loss of function and / or irreversible damage to the affected tissue or organ occurs.

[0048] In the sense of this patent application, the terms "lesion," "microlesion," and "trauma" refer to damage of different sizes and extents in the affected lung tissue. They can be caused by spontaneous physical impacts in which impact or rotational forces result in tissue damage. However, they can also be the final result of a previous degenerative disease of the affected lung tissue, or conversely, a microlesion can be the starting point of such a degenerative disease that follows the microlesion. Also, inflammation of the affected lung tissue can be favorable to such microlesions or trauma, or can be a sequela thereof. Therefore, these terms are interrelated with inflammation and degenerative disease.

[0049] In the sense of this patent application, the term "primary" disease, for example as "primary inflammatory or degenerative disease", refers to a lung disease that is not autoimmune mediated.

[0050] If it is known that a healthy individual suffers from or is susceptible to an inflammatory or degenerative disease, or that tissue damage is expected due to a certain excessive strain on the respective tissue or organ, it may be indicated to administer a prophylactic agent to prevent or at least alleviate the expected injury or damage. Therefore, the present patent application also refers to the prophylactic use according to the present invention.

[0051] Inflammatory lung diseases may also lead to degenerative diseases. Therefore, further examples are provided below. Therefore, the present patent application refers to the use according to the present invention in the prevention and / or treatment of inflammatory and / or degenerative lung diseases, in particular in the treatment of primary inflammatory and / or degenerative lung diseases.

[0052] Within the scope of this application, the term "pulmonary" refers to organs and tissues of the lower respiratory tract. Examples of organs and tissues of the lower respiratory tract include, but are not limited to, the lungs, including the lobes, apex, ligules, and alveoli; the bronchi, including the respiratory bronchioles; the tracheal and bronchial rings, including the carina; the pulmonary and bronchial vessels, including the lung and bronchial vessels; the bronchopulmonary lymph nodes; and the pulmonary autonomic nervous system.

[0053] Within the scope of this application, "pulmonary" also refers to adjacent organs and tissues that are functionally or structurally closely related to the lower respiratory tract and / or thorax and therefore have excellent pharmaceutical accessibility via inhalation. Examples include, but are not limited to, the pleura and diaphragm.

[0054] Within the scope of this application, the terms "alveoli" and "alveolar" refer to the tissue structures at the base of the pulmonary airways. Alveoli are hollow, cup-shaped cavities found in the lung parenchyma where gas exchange takes place. They are sparsely located on the respiratory bronchioles, line the walls of the alveolar ducts, and are more numerous in the blind-ended alveolar sacs. The alveolar membrane is the gas exchange surface surrounded by a network of capillaries. Oxygen diffuses across the membrane into the capillaries, and carbon dioxide is released from the capillaries into the alveoli to be exhaled. Alveoli consist of an epithelial layer of simple squamous epithelium and an extracellular matrix surrounded by capillaries. The epithelial lining is part of the alveolar membrane, also known as the respiratory membrane.

[0055] Type I and type II pneumocytes are found in the walls of the alveoli. Alveolar macrophages are immune cells that move within the alveolar lumen and the connective tissue between them. Type I cells are squamous epithelial cells, thin and flat, that form the structure of the alveoli. Type II cells (goblet cells) release pulmonary surfactant to reduce surface tension.

[0056] A typical paired human lung contains approximately 300 million alveoli and 70 m 2 Each alveolus is surrounded by a fine mesh of capillaries that cover approximately 70% of its area. The diameter of a typical healthy alveolus is 200-500 μm.

[0057] In Example 1, ex vivo mouse lungs were exposed to cigarette smoke for 5 minutes. Cigarette smoke is known to contain numerous cytotoxic agents, resulting in a dramatic intracellular increase in reactive oxygen species (ROS) and reactive nitrogen species (RNS) in affected lung tissue (a.o.). ROS / RNS are known to cause multiple cell damages, such as radical formation of additional cellular molecules, peroxide formation, lipid peroxidation, damage to cell and intracellular membranes, DNA damage, undesirable protein modifications, and induction of apoptosis. On the other hand, they are important mediators for initiating immune responses that help cells, tissues, or entire organisms deal with xenobiotics (e.g., toxins derived from cigarette smoke) and infection. The problem is an overshoot of the immune response, which causes the aforementioned symptoms of acute or chronic inflammation. Therefore, reducing excessive intracellular ROS / RNS levels is generally recognized as a promising approach for anti-inflammatory therapy, particularly for inflammatory lung diseases. Therefore, the ex vivo lung model used in Example 1 not only demonstrates therapeutic efficacy in cigarette smoke-induced lung diseases such as COPD, but also in all inflammatory lung diseases.

[0058] Administration of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt to the cigarette smoke-stimulated mouse lungs showed a dose-dependent reduction in ROS / RNS levels to pre-cigarette smoke control levels, demonstrating that 5-amino-2,3-dihydro-1,4-phthalazinedione, and particularly 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, has great potential for preventive and therapeutic efficacy in inflammatory lung diseases.

[0059] Inflammatory lung diseases can be classified as follows (ICD-10 Chapter X: Diseases of the respiratory system (J00-J99), Version 2016, as of January 10, 2020):

[0060] a) Inflammation of the lower respiratory tract due to bacterial, viral, fungal or parasitic infection These diseases include, but are not limited to, influenza caused by identified avian influenza viruses; influenza with pneumonia (identified influenza viruses); influenza with other respiratory symptoms (identified influenza viruses); influenza with other symptoms (identified influenza viruses); influenza with pneumonia (no virus identified); influenza with other respiratory symptoms (no virus identified); influenza with other symptoms (no virus identified); adenovirus pneumonia; Streptococcus pneumoniae pneumonia; Haemophilus influenzae pneumonia; Klebsiella pneumoniae pneumonia; Pseudomonas pneumonia; Staphylococcus pneumonia; Streptococcus, group B pneumonia; other streptococcal pneumonia; Escherichia coli pneumonia; Includes pneumonia due to coli; pneumonia due to other aerobic gram-negative bacteria; pneumonia due to Mycoplasma pneumoniae; other bacterial pneumonia; bacterial pneumonia (unspecified); chlamydial pneumonia; pneumonia due to other specified infectious organisms; pneumonia in bacterial diseases classified elsewhere; pneumonia in viral diseases classified elsewhere; pneumonia in fungal diseases; pneumonia in parasitic diseases; pneumonia in other diseases classified elsewhere; bronchopneumonia (unspecified); lobar pneumonia (unspecified); low-proliferative pneumonia (unspecified); other pneumonia (organism unspecified); pneumonia (unspecified); acute bronchitis; acute bronchiolitis; and unspecified acute lower respiratory tract infection.

[0061] b) Chronic lower respiratory tract disease These diseases include, but are not limited to, bronchitis not specified as acute or chronic; simple and mucopurulent chronic bronchitis; chronic bronchitis; chronic tracheitis; chronic tracheobronchitis; emphysema; chronic obstructive pulmonary disease (COPD); asthma; asthmatic conditions; bronchiectasis; pulmonary sarcoidosis; and alveolar microlithiasis.

[0062] c) Pulmonary diseases caused by external agents These diseases include, but are not limited to, coal workers' pneumoconiosis; asbestosis; talc dust pneumoconiosis; silicosis; aluminum lung disease; bauxite pulmonary fibrosis; beryllium lung disease; graphite pulmonary fibrosis; iron deposition disease; tin deposition disease; other specified inorganic dust pneumoconiosis; unspecified pneumoconiosis; pneumoconiosis associated with tuberculosis; byssinosis; linen wear disease; hemp fibrosis; other specified organic dust respiratory tract disease; farmer's lung; sugarcane lung disease; poultry farmer's disease; cork lung; malt worker's lung; mushroom grower's lung; maple bark processor's lung; air-conditioning and humidifier lung; cheesemaker's lung; coffee worker's lung; fishmeal-worker's lung; furrier's lung Includes other organic dust hypersensitivity pneumonitis such as allergic alveolitis and hypersensitivity pneumonitis; unspecified organic dust and hypersensitivity pneumonitis such as allergic alveolitis and hypersensitivity pneumonitis; respiratory conditions due to inhalation of chemicals, gases, smoke, and vapors; solid and liquid pneumonitis, radiation pneumonitis, post-irradiation pulmonary fibrosis, acute drug-induced interstitial lung injury; chronic drug-induced interstitial lung injury; drug-induced interstitial lung injury (unspecified); other specified external agent-induced respiratory conditions; and unspecified external agent-induced respiratory conditions.

[0063] d) Respiratory diseases that primarily affect the interstitium These diseases include, but are not limited to, adult respiratory distress syndrome; pulmonary edema, such as cardiogenic pulmonary edema, pulmonary permeability fluid, and high-altitude pulmonary edema; eosinophilic asthma; Loffler's pneumonia; tropical pulmonary eosinophilia; alveolar and mural alveolar conditions, Hamman-Rich syndrome; pulmonary fibrosis; idiopathic pulmonary fibrosis; other specified interstitial lung diseases; and interstitial lung disease (unspecified).

[0064] e) Suppuration and / or necrosis of the lower respiratory tract These diseases include, but are not limited to, pneumonia, pneumothorax, and lung abscesses with abscesses.

[0065] f) Pleural disease These conditions include, but are not limited to, pleurisy with pleural effusion; pleural effusion as classified elsewhere; pleural plaque; pneumothorax; chylothorax; fibrothorax; hemothorax; hemopneumothorax; hydrothorax; and pleural conditions (unspecified).

[0066] g) Postoperative or related lower respiratory tract illness These conditions include, but are not limited to, acute pulmonary dysfunction after thoracic surgery; acute pulmonary dysfunction after non-thoracic surgery; chronic pulmonary dysfunction after surgery; host-versus-graft disease after lung transplantation; graft-versus-host disease after lung transplantation; chronic pulmonary allograft dysfunction (CLAD); chronic pulmonary allograft dysfunction-bronchiolitis obliterans syndrome (CLAD-BOS); pulmonary ischemia-reperfusion injury; primary graft dysfunction after lung transplantation; Mendelsohn's syndrome; other postoperative respiratory disorders; postoperative respiratory disorders (unspecified); respiratory failure (not elsewhere classified); bronchial disease (not elsewhere classified); lung collapse; atelectasis; interstitial emphysema; pneumomediastinum; compensated emphysema; mediastinitis; and diaphragmatic disorders.

[0067] h) Perinatal-specific lung diseases These diseases include, but are not limited to, neonatal respiratory distress syndrome; transient tachypnea of ​​the newborn; congenital pneumonia caused by viral agents; congenital pneumonia caused by Chlamydia; congenital pneumonia caused by Staphylococcus; congenital pneumonia caused by Streptococcus group B; congenital pneumonia caused by Escherichia coli; congenital pneumonia caused by Pseudomonas; congenital pneumonia caused by Haemophilus influenzae, Klebsiella pneumoniae, and other pathogens. pneumoniae, Mycoplasma, and non-group B streptococci; congenital pneumonia due to other organisms; unspecified congenital pneumonia; neonatal meconium aspiration; perinatal interstitial emphysema; perinatal pneumothorax; perinatal emphysema mediastinum; other conditions associated with perinatal interstitial emphysema; perinatal pulmonary hemorrhage; Wilson-Mikity syndrome; bronchopulmonary dysplasia of perinatal origin; and unspecified chronic respiratory disease of perinatal origin.

[0068] i) Trauma and injury to the lower respiratory tract and / or thoracic cage These conditions include, but are not limited to, injuries to the pulmonary blood vessels, traumatic pneumothorax; traumatic hemothorax; traumatic hemopneumothorax; other injuries to the lungs; bronchial injuries; pleural injuries; diaphragmatic injuries; chest crush injuries; and traumatic amputation of a portion of the chest.

[0069] j) Malignant neoplasms of the lower respiratory tract These diseases include, but are not limited to, malignant neoplasms of the bronchi and lung; lung carcinoma; non-small cell lung carcinoma; adenocarcinoma; squamous cell lung carcinoma; large cell lung carcinoma; pulmonary intestinal adenocarcinoma; bronchioloalveolar carcinoma; ovine lung adenocarcinoma; small cell lung carcinoma; endobronchial leiomyoma; bronchial carcinoma; Pancoast tumor; pulmonary carcinoid tumor; pleuropulmonary blastoma; neuroendocrine tumors of the lung; pulmonary lymphoma; pulmonary lymphangiomatosis; pulmonary sarcoma; alveolar soft part sarcoma; vascular tumors of the lung; mediastinal tumors; pleural tumors; and metastases to the lung.

[0070] In particular, the present application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, the inflammatory lung diseases being selected from the group comprising inflammation of the lower respiratory tract due to bacterial, viral, fungal or parasitic infection, chronic lower respiratory tract diseases, lung diseases due to external agents, respiratory diseases mainly affecting the interstitium, suppurative and / or necrotic conditions of the lower respiratory tract, pleural diseases, post-operative or related lower respiratory tract diseases, perinatal-specific lung diseases, trauma and injury of the lower respiratory tract and / or thorax, and malignant neoplasms of the lower respiratory tract.

[0071] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, the inflammatory lung diseases being inflammation of the lower respiratory tract due to bacterial, viral, fungal or parasitic infection.

[0072] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, wherein the inflammatory lung diseases are chronic lower respiratory tract diseases.

[0073] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, the inflammatory lung diseases being lung diseases caused by external agents.

[0074] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, which inflammatory lung diseases are respiratory diseases that primarily affect the interstitium.

[0075] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, the inflammatory lung diseases being suppurative and / or necrotic conditions of the lower respiratory tract.

[0076] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, wherein the inflammatory lung diseases are pleural diseases.

[0077] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, the inflammatory lung diseases being post-operative or associated lower respiratory tract diseases.

[0078] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, the inflammatory lung diseases being perinatal-specific lung diseases.

[0079] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, the inflammatory lung diseases being conditions resulting from trauma and / or injury of the lower respiratory tract and / or thorax.

[0080] In particular, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, the inflammatory lung diseases being malignant neoplasms of the lower respiratory tract.

[0081] Of particular interest within the scope of this application is COPD, which is a progressive development of airflow limitation that is not fully reversible. Most COPD patients suffer from three pathological conditions: bronchitis, emphysema, and mucus obstruction. This disease is characterized by a slowly progressive, irreversible decrease in forced expiratory volume in the first second of expiration (FEV1) with a relatively preserved forced vital capacity (FVC). In both asthma and COPD, there is significant but distinct remodeling of the airways. The majority of airflow obstruction is due to two major components: alveolar destruction (emphysema) and small airway obstruction (chronic obstructive bronchitis). COPD is primarily characterized by marked mucus cell hyperplasia. A key feature of COPD is neutrophil infiltration into the lungs of patients. Elevated levels of pro-inflammatory cytokines such as TNF-alpha, and particularly chemokines such as interleukin-8 (IL-8), play a prominent role in the pathogenesis of COPD. Platelet thromboxane synthesis has been found to be enhanced in COPD patients. Much of the tissue damage is caused by neutrophil activation and the subsequent release of matrix metalloproteinases and increased production of ROS and RNS.

[0082] Emphysema represents the destruction of lung structure, with enlargement of air spaces and loss of alveolar surface area. Lung damage is caused by weakening and destruction of the air sacs within the lungs. Several adjacent alveoli may rupture, forming one large space rather than many smaller ones. The larger spaces can become even larger cavities called bullae. As a result, the natural elasticity of lung tissue is lost, leading to excessive stretching and rupture, thus minimizing lung compliance. There is also less traction on the small bronchi, which can cause them to collapse and obstruct airflow. Air not exhaled before the next respiratory cycle becomes trapped in the lungs, leading to shortness of breath. The enormous effort required to push air out of the lungs during exhalation leaves the patient exhausted.

[0083] The most common symptoms of COPD include shortness of breath, chronic cough, chest tightness, increased effort to breathe, increased mucus production, and frequent throat clearing, which can prevent patients from carrying out normal daily activities.

[0084] Long-term smoking is the most common cause of COPD, accounting for 80-90% of all cases. Other risk factors are genetics, passive smoking, air pollution, and a history of frequent childhood respiratory infections. COPD is progressive and sometimes irreversible, and there is currently no cure.

[0085] Clinical development in COPD is typically described in three stages.

[0086] Stage 1: Lung function (measured by FEV1) is greater than or equal to 50% of predicted normal lung function. There is minimal impact on health-related quality of life. Symptoms may progress at this stage, and patients may begin to experience severe shortness of breath and require evaluation by a pulmonologist.

[0087] Stage 2: FEV1 lung function is 35-49% of predicted normal lung function, with a significant impact on health-related quality of life.

[0088] Stage 3: FEV1 lung function is less than 35% of predicted normal lung function, with a significant impact on health-related quality of life.

[0089] Symptomatic treatments include the administration of bronchodilators, glucocorticoids, and PDE4 inhibitors. Suitable bronchodilators include, for example, beta-2 adrenergic agonists such as short-acting fenoterol and salbutamol, and long-acting salmeterol and formoterol, muscarinic anticholinergics such as ipratropium bromide and tiotropium bromide, and methylxanthines such as theophylline.

[0090] Suitable glucocorticoids include inhaled glucocorticoids such as budesonide, beclomethasone and fluticasone, orally administered glucocorticoids such as prednisolone, and intravenously administered glucocorticoids such as prednisolone.

[0091] A suitable PDE (phosphodiesterase) 4 inhibitor is roflumilast.

[0092] Therefore, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of COPD by inhalation administration.

[0093] Of particular interest within the scope of this application is asthma, a chronic inflammatory disease of the lower respiratory tract. It is primarily characterized by recurrent symptoms such as reversible airflow obstruction and easily provoked bronchospasm. Symptoms include episodes of wheezing, coughing, chest tightness, and difficulty breathing.

[0094] Asthma is thought to be caused by a combination of genetic and environmental factors, including air pollution and exposure to allergens. Other potential triggers may be iatrogenic.

[0095] Asthma is clinically classified into intermittent, mild persistent, moderate persistent and severe persistent based on the frequency of symptoms. The most important parameters are FEV1 and peak expiratory flow.

[0096] To date, asthma has not been curable. For long-term treatment, symptoms such as asthma can be pharmacologically prevented by avoiding triggers such as allergens and irritants and by using inhaled corticosteroids. Long-acting beta-2 agonists (LABAs) or anti-leukotrienes may also be used. The most common inhaled corticosteroids include beclomethasone, budesonide, fluticasone, mometasone, and ciclesonide. Suitable LABAs include salmeterol and formoterol. Orally administered leukotriene receptor antagonists, such as montelukast, pranlukast, and zafirlukast, are used. Suitable 5-lipoxygenase (5-LOX) inhibitors include meclofenamate sodium and zileuton. In severe stages of asthma, intravenous corticosteroids, such as prednisolone, are recommended.

[0097] Acute asthma attacks (status asthmaticus) are best treated with inhaled short-acting beta-2 agonists such as salbutamol. Ipratropium bromide can also be inhaled. Intravenous corticosteroids can be given.

[0098] Inhalation administration is generally via a metered dose inhaler, respectively a dry powder inhaler.

[0099] Therefore, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof in the prevention or treatment of asthma by inhalation administration.

[0100] Pulmonary sarcoidosis is also of particular interest within the scope of this application. Pulmonary sarcoidosis (used interchangeably herein: pulmonary sarcoidosis, PS) is characterized by abnormal collections of inflammatory cells in the lungs, forming masses known as granulomas. The cause of sarcoidosis is unknown. The disease usually begins in the lungs, skin, or lymph nodes and can manifest throughout the body. The most common symptom is long-lasting fatigue, even if disease activity ceases. General malaise, shortness of breath, joint complaints, elevated body temperature, weight loss, and skin complaints may be present. The prognosis is generally good. Acute forms, in particular, usually cause few problems because the symptoms gradually subside naturally. If sarcoidosis is present in the heart, kidneys, liver, and / or central nervous system, or is widespread in the lungs, the outcome is less favorable. Sarcoidosis is generally classified into four stages, determined by chest radiography. However, these stages do not correlate with severity. 1. Hilar lymphadenopathy (granulomas in the lymph nodes); 2. Hilar lymphadenopathy and reticulonodular infiltrates (pulmonary granulomas); 3. Bilateral pulmonary infiltrates (granulomas in the lungs but not in the lymph nodes); 4. Fibrocystic sarcoidosis (irreversible scarring in the lungs, i.e., pulmonary fibrosis), typically with upward-sloping hilar recession, cystic, and bullous changes. Patients in stages 2 and 3 often have a chronic progressive disease course.

[0101] Symptomatic treatments for pulmonary sarcoidosis include corticosteroids such as prednisone and prednisolone, immunosuppressants such as TNF-alpha inhibitors (etanercept, adalimumab, golimumab, infliximab), cyclophosphamide, cladribine, cyclosporine, chlorambucil, and chloroquine, IL-23 inhibitors such as tildrakizumab and guselkumab, and antimetabolites such as mycophenolate, leflunomide, azathioprine, and methotrexate. In subtypes such as Lofgren's syndrome, COX inhibitors such as acetylsalicylic acid, diclofenac, or ibuprofen are used. All of these active agents have been administered systemically to date.

[0102] Therefore, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of pulmonary sarcoidosis by inhalation administration.

[0103] Cystic fibrosis is also of particular interest within the scope of this application. Cystic fibrosis is an inherited form of chronic bronchitis accompanied by mucus hypersecretion, generally associated with poor clearance of airway secretions, airflow obstruction, and chronic bacterial infection of the airways, commonly caused by Pseudomonas aeruginosa. It is known that sputum and bronchoalveolar lavage fluid from cystic fibrosis patients reduce the ability of neutrophils to kill these bacteria. Airway obstruction by such secretions can cause respiratory distress and, in some cases, can lead to respiratory failure and death. Additional symptoms include sinus infections, poor growth, steatorrhea, clubbing of fingers and toes, and male infertility.

[0104] Cystic fibrosis is an autosomal recessive disease caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein. CFTR is involved in the production of sweat, digestive juices, and mucus. CFTR also regulates the transport of H2O and Cl2 into and out of cells within the lungs. - CFTR is a channel protein that controls the flow of ions. When the CFTR protein is functioning properly, ions flow freely in and out of cells. However, when the CFTR protein is dysfunctional, these ions cannot leave the cell due to the blocked channel. This causes cystic fibrosis, a condition characterized by the accumulation of thick mucus in the lungs.

[0105] There is no known cure for cystic fibrosis. Intravenous, inhaled, and oral antibiotics are used to treat chronic and acute infections. Mechanical devices and inhaled medications are used to alter and remove thickened mucus. These treatments, while effective, can be very time-consuming. Home oxygen therapy is recommended for people with significantly low oxygen levels. Inhaled antibiotics include levofloxacin, tobramycin, aztreonam, and colistin. Orally administered antibiotics include ciprofloxacin and azithromycin. Mutation-specific CFTR enhancers include ivacaftar and tezacaftar.

[0106] Thus, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of cystic fibrosis by inhaled administration.

[0107] Bronchiectasis is also of particular interest within the scope of this application. Bronchiectasis is considered an idiopathic disease. Morphologically, it is characterized by permanent enlargement of a segment of the lower respiratory tract. Pathological conditions considered include, for example, post-infectious, immunodeficiency, exaggerated immune response, congenital anomalies, inflammatory pneumonia, fibrosis, and mechanical obstruction. Symptoms include chronic cough with daily mucus production. Therefore, it resembles cystic fibrosis, but without the characteristic genetic mutation. Pulmonary function test results generally show airflow obstruction ranging from moderate to severe. Additional symptoms include dyspnea, coughing up blood, chest pain, hemoptysis, fatigue, and weight loss.

[0108] Treatment of bronchiectasis aims to control infection and bronchial secretions, relieve airway obstruction, and remove affected lung segments through surgery or arterial embolization. Antibiotics, particularly macrolide antibiotics, are administered as needed. Excessive mucus production can be addressed with mucolytics. Bronchodilators are used to facilitate breathing. Continuous inhaled corticosteroids reduce phlegm production, decrease airway narrowing, and provide some protection against disease progression.

[0109] Therefore, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof in the prevention or treatment of bronchiectasis by inhalation administration.

[0110] Adult respiratory distress syndrome (ARDS) is also of particular interest within the scope of this application. Adult respiratory distress syndrome (ARDS) is a respiratory insufficiency syndrome. It can have various causes, such as pneumonia, trauma, severe burns, blood transfusion, aspiration, sepsis, pancreatitis, or can occur as a reaction to certain drugs. Gas exchange within the alveoli is severely impaired due to alveolar endothelial cell injury, surfactant dysfunction, an overshooting immune system response, and coagulopathy. Rapid migration of neutrophils and T lymphocytes into affected lung tissue is observed. Acute symptoms include dyspnea, tachypnea, and pale skin. If patients survive, pulmonary function is often permanently impaired. Acute treatment is primarily based on mechanical ventilation in the intensive care unit and, if necessary, the administration of antibiotics. Nitric oxide inhalation can help improve blood oxygenation but has other drawbacks. Extracorporeal membrane oxygenation (ECMO) helps increase survival rates. However, pharmaceutical treatment, for example with corticosteroids, remains controversial.

[0111] Accordingly, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of adult respiratory distress syndrome by inhalation administration.

[0112] Pulmonary fibrosis is also of particular interest within the scope of this application. Scarring occurs in lung tissue, leading to serious respiratory problems. Scarring, respectively, and the accumulation of excess fibrous connective tissue, leads to thickening of the lung walls and thus to a reduction in oxygen supply in the blood. The result is chronic, progressive respiratory distress. Pulmonary fibrosis is often secondary to other lung diseases, such as interstitial lung disorders, autoimmune lung diseases, inhalation of environmental and occupational pollutants, or certain infectious diseases. Others are classified as idiopathic pulmonary fibrosis.

[0113] Pulmonary fibrosis involves the gradual replacement of lung parenchyma with fibrous tissue. Scar tissue causes an irreversible decrease in oxygen diffusing capacity, resulting in decreased lung stiffness or compliance. Pulmonary fibrosis is sustained by abnormal wound healing.

[0114] To date, there is no general pharmaceutical treatment for pulmonary fibrosis. Some subtypes respond to corticosteroids such as prednisone, antifibrotic agents such as pirfenidone and nintedanib, or immunosuppressants such as cyclophosphamide, azathioprine, methotrexate, penicillamine, and cyclosporine.

[0115] Therefore, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of pulmonary fibrosis, respectively idiopathic pulmonary fibrosis, by inhalation administration.

[0116] A typical inflammatory disease caused by external agents is beryllium pulmonary disease (interchangeably referred to herein as chronic beryllium disease, CBD). There is no cure for this occupational disease; only symptomatic treatment is available.

[0117] Prolonged exposure via inhalation can sensitize the lungs to beryllium, leading to the development of small inflammatory nodules called granulomas. Typically, CBD granulomas are not characterized by necrosis and therefore do not have a caseous appearance. Ultimately, this process leads to a decrease in the lung's diffusing capacity. Typical symptoms are cough and dyspnea. Other symptoms include chest pain, joint pain, weight loss, and fever. The patient's T cells become sensitized to beryllium. A pathological immune response leads to the accumulation of CD4+ helper T lymphocytes and macrophages in the lungs, where they aggregate together to form granulomas. Ultimately, this leads to pulmonary fibrosis. Treatment options include supplemental oxygen and oral corticosteroids.

[0118] Accordingly, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of beryllium pulmonary disease by inhalation administration.

[0119] Chronic lung allograft dysfunction is also of particular interest within the scope of this application. Chronic lung allograft dysfunction (CLAD), and respectively chronic lung allograft dysfunction-bronchiolitis obliterans syndrome (CLAD-BOS), are major problems in the long-term management of lung transplant recipients. Both alloimmune-dependent (rejection) and alloimmune-independent factors contribute to the development of CLAD. It encompasses all forms of chronic pulmonary function decline after exclusion of known causes (persistent acute rejection, infection, anastomotic stenosis or disease recurrence, pleural disease, diaphragmatic dysfunction, or native lung hyperinflation). Thus, it is a heterogeneous entity in which two major phenotypes have been identified: bronchiolitis obliterans syndrome (BOS), defined by a persistent decline in FEV1 and an obstructive functional pattern.

[0120] There is currently no treatment available to reverse CLAD after diagnosis. Pharmacological treatment of symptoms involves azithromycin (first-line treatment) or montelukast. In treatment-resistant cases, photopheresis is indicated.

[0121] Therefore, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of CLAD, respectively CLAD-BOS, by inhaled administration.

[0122] Pulmonary edema is also of particular interest within the scope of this application. Pulmonary edema can have different causes. Fluid accumulation occurs in the tissues and air spaces of the lungs, leading to impaired gas exchange and, in the worst cases, respiratory failure. Treatment of pulmonary edema is primarily focused on maintaining vital functions, for example, by tracheal intubation and mechanical ventilation. Hypoxic symptoms can be addressed by supplemental oxygen.

[0123] Cardiogenic pulmonary edema can result from congestive heart failure, which causes the heart to be unable to pump blood through the pulmonary circulation at a sufficient rate, resulting in increased wedge pressure and pulmonary edema. The underlying cause can be left ventricular failure, arrhythmia, or fluid overload due to, for example, renal failure or intravenous therapy. It can also be caused by hypertensive crisis, as elevated blood pressure and increased afterload on the left ventricle impede forward flow, causing increased wedge pressure and subsequent pulmonary edema. In acute cases, loop diuretics such as furosemide are often administered along with morphine to relieve respiratory distress. While both diuretics and morphine can have vasodilatory effects, certain nitric oxide vasodilators, such as intravenous glyceryl trinitrate or isosorbide dinitrate, can also be used.

[0124] Pulmonary permeability water species include alveolar Na + It is characterized by reduced uptake capacity and impaired capillary barrier function and is a potentially fatal complication, for example, in listeriolysin-induced listeriosis. + Uptake is primarily mediated by the epithelial sodium channel (ENaC) and initiates alveolar fluid clearance.

[0125] High-altitude pulmonary edema (HAPE) typically occurs in otherwise healthy people at altitudes above 2,500 meters and can be life-threatening. After rapid elevation gain, symptoms may include shortness of breath at rest, cough, weakness or decreased exercise capacity, chest tightness or congestion, crackles or wheezing, central blue skin color, tachypnea, and tachycardia. Arterial oxygen tension decreases due to lower atmospheric pressure at high altitudes. Increased capillary pressure occurs due to hypoxemic pulmonary hypertension secondary to hypoxic pulmonary vasoconstriction. This leads to subsequent leakage of cells and proteins into the alveoli. Hypoxic pulmonary vasoconstriction occurs diffusely, resulting in arterial vasoconstriction in all regions of the lungs.

[0126] The first medical action is to descend to a lower altitude as quickly as possible. pO2Supplemental oxygen can be administered to maintain a blood oxygen level above 90%.

[0127] Pharmacological prevention of HAPE includes calcium channel blockers such as nifedipine, PDE5 inhibitors such as sildenafil and tadalafil, and inhaled beta-2 agonists such as salmeterol.

[0128] A new pharmaceutical approach to enhancing ENaC function is, for example, the peptide drug solnatide.

[0129] Therefore, the present application also refers to the use of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of pulmonary edema by inhalation administration, in particular for the prevention or treatment of cardiogenic pulmonary edema, pulmonary permeability water and high altitude pulmonary edema.

[0130] Pulmonary ischemia-reperfusion injury is also of particular interest within the scope of this application. In lung transplantation, organ ischemia and subsequent reperfusion are inevitable, resulting in acute post-transplant sterile inflammation commonly referred to as ischemia-reperfusion (IR) injury. Severe IR injury leads to primary graft dysfunction (PGD), which is a major cause of both short- and long-term morbidity and mortality after lung transplantation. Currently, no therapeutic agents are clinically available to specifically prevent IR injury, and treatment strategies are limited to supportive care. When feasible, donor lungs, respectively, can be prophylactically treated with 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts.

[0131] Endothelial dysfunction and disruption of the endothelial barrier are hallmarks of pulmonary IR injury. Endothelial membrane depolarization induces ROS production and subsequent inflammation and leukocyte emigration. Activation of NADPH oxidase (NOX2), induction of nitric oxide (NO) production, and activation of integrin αvβ5 promote vascular permeability through ROS / RNS production. Alveolar macrophages are activated. Increased chemokine concentrations and adhesion molecule expression on endothelial cells and neutrophils lead to neutrophil binding and infiltration, releasing cytokines, ROS, and forming neutrophil extracellular traps (NETs).

[0132] Recent preventive strategies before lung transplantation include administering antioxidants (free radical scavengers) or inhibitors of oxidant-generating enzymes (e.g., methylene blue or N-acetylcysteine) to the organ recipient, anti-inflammatory strategies using inhibitors of pro-inflammatory transcription factors or inflammatory mediators, ventilation with gas molecules such as carbon monoxide or the inhaled anesthetic sevoflurane, growth factors or nutritional supplements such as creatine, and cell-based therapies such as the administration of mesenchymal stem cells.

[0133] Therefore, the present application also refers to the use of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof in the prevention or treatment of pulmonary ischemia-reperfusion injury by inhalation administration.

[0134] Of particular interest within the scope of this application is primary graft dysfunction after lung transplantation. Primary graft dysfunction (PGD) is a devastating form of acute lung injury that affects approximately 10% to 25% of patients in the first few hours to days after lung transplantation. Clinically and pathologically, it mimics adult respiratory distress syndrome (ARDS) and is associated with a mortality rate of up to 50%. PGD can have different causes, including the aforementioned ischemia-reperfusion injury, epithelial cell death, endothelial cell dysfunction, innate immune activation, oxidative stress, release of inflammatory cytokines and chemokines, and iatrogenic factors such as mechanical ventilation and transfusion of blood components. Activation of the innate immune system has been demonstrated during the development and progression of ischemia-reperfusion injury. Herein, PGD is associated with the innate immune pathway of Toll-like receptor-mediated injury.

[0135] Molecular markers of PGD include intercellular adhesion molecule-1, surfactant protein-1, plasminogen activator inhibitor, soluble receptor for advanced glycation end products, and protein C.

[0136] Approaches to avoid PGD include optimizing reperfusion, adjusting prostaglandin concentrations, hemodynamic control, hormone replacement, ventilator management, and donor lung preparation strategies. To reduce the incidence of PGD, strategies such as the use of prostaglandins, nitric oxide, surfactants, and adenosine, or inhibition of proinflammatory mediators and / or scavenging free oxygen radicals have been used. Furthermore, inhibitors of free oxygen radicals, cytokines, proteases, lipid mediators, adhesion molecules, and complement cascades have been investigated for the inhibition of neutrophils and neutrophil-mediated mediators. Inhaled nitric oxide can reduce pulmonary artery pressure without affecting systemic blood pressure. As a last-ditch lifesaving option, extracorporeal membrane oxygenation (ECMO) is used to correct PGD-induced hypoxemia and provide the necessary gas exchange.

[0137] Therefore, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of primary graft dysfunction after lung transplantation by inhalation administration.

[0138] For effective prophylactic or therapeutic treatment of the aforementioned inflammatory lung diseases, 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof must reach the patient's alveoli. Therefore, the particle size must be small enough to reach the lowest part of the airways of the lung tissue. The best class of inhalation device for inhalation administration of pharmaceutically active agents is the so-called mesh nebulizer mentioned above. Within the scope of this application, virtually all mesh nebulizers known in the art can be used, from rather simple disposable mesh nebulizers for cough and cold or recreational use to sophisticated high-end mesh nebulizers for clinical or home treatment of serious diseases or conditions of the lower respiratory tract.

[0139] Suitable commercially available mesh nebulizers, jet nebulizers, ultrasonic nebulizers, dry powder inhalers and (pressurized) metered dose inhalers include, but are not limited to, eFlow® rapid, PARI LC STAR®, PARI Velox and PARI Velox Junior (PARI GmbH, Starnberg, Germany), Philips Respironics I-neb and Philips InnoSpire Go (Koninklijke Philips NV, Eindhoven, The Netherlands), VENTA-NEB®-ir, OPTI-NEB®, M-neb® dose. +Mesh nebulizer inhalation MN-300 / 8, M-Neb Flow+ and M-neb® Mesh nebulizer MN-300 / X (NEBU-TEC, Einsfeld, Germany), Hcmed Deepro HCM-86C and HCM860 (HCmed Innovations Co., Ltd., Taipei, Taiwan), OMRON MicroAir U22 and U100 (OMRON, Kyoto, Japan), Aerogen® Solo, Aerogen® Ultra and Aerogen® PRO (Aerogen, Galway, Ireland), KTMED NePlus NE-SM1 (KTMED Inc., Seoul, Korea), Vectura Bayer Breelib™ (Bayer AG, Leverkusen, Germany), Vectura Fox, MPV Truma and MicroDrop® Smarty (MPV MEDICAL GmbH, Kirchheim, Germany), MOBI MESH (APEX Medical, New Taipei City, Taiwan), B.Well WN-114, TH-134 and TH-135 (B.Well Swiss AG, Widnau, Switzerland), Babybelle Asia BBU01 (Babybelle Asia Ltd., Hong Kong), CA-MI Kiwi and others (CA-MI sri, Langhirano, Italy), Diagnosis PRO MESH (Diagnosis SA, Bialystok, Poland), DIGI O2 (DigiO2 International Co., Ltd., New Taipei City, Taiwan), feellife AIR PLUS, AEROCENTRE+, AIR 360+, AIR GARDEN, AIRICU, AIR MASK, AIRGEL BOY, AIR ANGEL, AIRGEL GIRL and AIR PRO 4 (Feellife Health Inc., Shenzhen, China), Hannox MA-02 (Hannox International Corp., Taipei, Taiwan), Health and Life HL100 and HL100A (HEALTH&LIFE Co., Ltd., New Taipei City, Taiwan), Honsun NB-810B (Honsun Co., Ltd.), Nantong, China), K-jump (registered trademark) KN-9100 (K-jump Health Co., Ltd., New Taipei City, Taiwan), microlife NEB-800 (Microlife AG, Widnau, Switzerland), OK Biotech Docspray (OK Biotech Co., Ltd., Hsinchu City, Taiwan), Prodigy Mini-Mist (registered trademark) (Prodigy Diabetes Care, LLC, Charlotte, USA), Quatek NM211, NE203, NE320 and NE403 (Big Eagle Holding Ltd., Taipei, Taiwan), Simzo NBM-1 and NBM-2 (Simzo Electronic Technology Ltd., Dongguan, China), Mexus(R) BBU01 and BBU02 (Tai Yu International Manufactory Ltd., Dongguan, China), TaiDoc TD-7001(TaiDoc Technology Co., New Taipei City, Taiwan), Vibralung® and HIFLO Miniheart Circulaire II (Westmed Medical Group, Purchase, USA), KEJIAN (Xuzhou Kejian Hi-Tech Co., Ltd., Xuzhou, China), YM-252, P&S-T45 and P&S-360 (TEKCELEO, Valbonne, France), Maxwell YS-31 (Maxwell India, Jaipur, India), Kernmed® JLN-MB001 (Kernmed, Durmersheim, Germany).

[0140] Mesh nebulizers with piezoelectric actuation of the nebulization process, respectively vibrating mesh nebulizers are preferred.

[0141] Mesh nebulizers can be divided into two groups based on their interaction with the patient: continuous-mode devices and trigger-activated devices. Continuous-mode mesh nebulizers continuously release nebulized aerosol into the mouthpiece, requiring the patient to inhale the aerosol. Trigger-activated devices release a defined amount of aerosol only during active, deep inspiration. Thus, a much larger volume of active-agent-containing aerosol is inhaled and reaches the lowest airways than continuous-mode devices. The latter lose a larger volume of active-agent-containing aerosol to either the periphery or the upper airway, since aerosol release is not coupled to the respiratory cycle.

[0142] Therefore, trigger-activated mesh nebulizers, especially vibrating mesh nebulizers, are preferred.

[0143] Particularly preferred are trigger-activated mesh nebulizers with piezoelectric actuation of the nebulization process.

[0144] Mesh nebulizer models PARI eFlow® rapid, Philips Respironics I-neb, Philips InnoSpire Go, M-neb® dose + Mesh nebulizer inhalation MN-300 / 8, Hcmed Deepro HCM-86C and HCM860, OMRON MicroAir U100, Aerogen® Solo, KTMED NePlus NE-SM1, Vectura Fox, Vectura Bayer Breelib™ are preferred.

[0145] The most preferred vibrating mesh nebulizer models are the PARI eFlow® rapid, PARI Velox, Philips Respironics I-neb, and M-neb® dose. +High-end models include mesh nebulizer inhalation MN-300 / 8, Aerogen® Solo, Vectura Fox, and Vectura Bayer Breelib®.

[0146] Therefore, the present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, wherein the mesh nebulizer releases an aerosol containing droplets of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for inhalation administration. This applies to the aforementioned subtypes of inflammatory lung diseases as well as to the aforementioned single inflammatory lung diseases, respectively.

[0147] The mean droplet size is usually characterized as the MMAD (mass aerodynamic diameter). The size of individual droplets is called the MAD (mass aerodynamic diameter). This value indicates the diameter of 50% or less of the nebulized particles (droplets). Particles with an MMAD > 10 μm usually do not reach the lower respiratory tract and often get stuck in the throat. Particles with an MMAD greater than 5 μm and less than 10 μm usually reach the bronchi but not the alveoli. Particles between an MMAD of 100 nm and 1 μm do not deposit in the alveoli and are immediately exhaled. Therefore, the optimal range is an MMAD of 1 μm to 5 μm. Recent publications further support a narrower range of 3.0 μm to 4.0 μm (see Amirav et al. (2010) J Allergy Clin Immunol 25:1206-1211; Haidl et al. (2012) Pneumologie 66:356-360).

[0148] Therefore, the MMAD of the nebulized 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts should be 3.0 μm to 4.0 μm, preferably 3.0 μm to 3.5 μm, more preferably 3.0 μm to 3.4 μm, even more preferably 3.0 μm to 3.3 μm, and most preferably 3.0 μm to 3.2 μm.

[0149] Similarly, at least 50% of the droplets of an aerosol according to the invention should have a MAD of 1 μm to 5 μm. Preferably, at least 50% of the droplets of an aerosol according to the invention should have a MAD of 1 μm to 4 μm, more preferably, at least 50% of the droplets of an aerosol according to the invention should have a MAD of 1 μm to 3.5 μm, and most preferably, at least 50% of the droplets of an aerosol according to the invention should have a MAD of 1 μm to 3.3 μm.

[0150] A further commonly accepted quality parameter is the percentage of particles in the generated aerosol with a diameter in the range of 1 μm to 5 μm (FPM; Fine Particle Mass). FPM is a measure of particle distribution. It is calculated by subtracting the percentage of particles in the generated aerosol with a diameter in the range below 1 μm from the total percentage of particles in the generated aerosol with a diameter in the range below 5 μm (FPF; Fine Particle Fraction).

[0151] Therefore, the FPM of the nebulized 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts must be at least 50%, preferably at least 55%, more preferably at least 60%, and most preferably at least 65%.

[0152] It is expressly stated that the values ​​and ranges disclosed herein refer to measurements of MMAD, FPF and FPM with a cascade impactor (Next Generation Impactor, NGI) cooled to 4°C.

[0153] To test whether the combination of a mesh nebulizer and 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt could meet this objective, the FPM in the aerosol thus generated was determined. As shown in Example 2, 5 ml of 20 mg / ml saline solution of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, prepared from Form I (see above), was first nebulized. Surprisingly, under these conditions, 69.0% of the particles (FPM) were found to be within the target range. This is a much more favorable particle size distribution percentage, as observed for many other pharmaceutically active agents. This percentage may, of course, vary slightly depending on the selected aqueous solution, temperature, mesh nebulizer model, selected piezoelectric excitation frequency, outlet geometry, and dosage per application. Furthermore, surprisingly, the MMAD for this nebulization was found to be 3.1 μm in the same experiment. Thus, this MMAD fully meets the desired range.

[0154] It is understood that the MMAD for this nebulization may vary slightly depending on conditions such as ambient temperature, the temperature of the pharmaceutical formulation being nebulized, the concentration of the pharmaceutically active agent, any choice of excipients, and the like.

[0155] The present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, wherein the mesh nebulizer emits an aerosol containing droplets of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for inhalation administration, wherein at least 50% of the droplets of the aerosol are in the size range of 1 μm to 5 μm in diameter.

[0156] In a more preferred embodiment, at least 55% of the droplets of the aerosol are in the size range of 1 μm to 5 μm in diameter.

[0157] In a more preferred embodiment, at least 60% of the droplets of the aerosol are in the size range of 1 μm to 5 μm in diameter.

[0158] In a particularly preferred embodiment, at least 65% of the droplets of the aerosol are in the size range of 1 μm to 5 μm in diameter.

[0159] The present application also refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, wherein the mesh nebulizer emits an aerosol containing droplets of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for inhalation administration, the droplets having a mass median aerodynamic diameter in the range of 3 μm to 4 μm.

[0160] In a preferred embodiment, the mass median aerodynamic diameter of these droplets is in the range of 3 μm to 3.5 μm.

[0161] In a more preferred embodiment, the mass median aerodynamic diameter of these droplets is in the range of 3 μm to 3.4 μm.

[0162] In an even more preferred embodiment, the mass median aerodynamic diameter of these droplets is in the range of 3 μm to 3.3 μm.

[0163] In a particularly preferred embodiment, the mass median aerodynamic diameter of these droplets is in the range of 3 μm to 3.2 μm.

[0164] Thus, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, wherein a mesh nebulizer emits an aerosol containing droplets of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for inhalation administration, the aerosol being characterized by a fine particle mass of at least 50% of the aerosol droplets.

[0165] Thus, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, wherein a mesh nebulizer emits an aerosol containing droplets of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for inhalation administration, the aerosol being characterized in that the aerosol droplets have an aerodynamic mass median diameter of 3.0 μm to 3.3 μm.

[0166] Accordingly, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of inflammatory lung diseases by inhalation administration, wherein a vibrating mesh nebulizer emits an aerosol containing droplets of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for inhalation administration, wherein the aerosol droplets have a mass median aerodynamic diameter of 3.0 μm to 3.3 μm, and the aerosol is characterized by a fine particle mass of at least 50% of the aerosol droplets, the fine particle mass representing the proportion of droplets having a diameter in the range of 1.0 to 5.0 μm.

[0167] In a preferred embodiment, the aerosol is characterized in that the aerosol droplets have a mass median aerodynamic diameter of 3.0 μm to 3.2 μm.

[0168] In another aspect, the application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in an aerosol for inhalation administration to prevent or treat inflammatory lung diseases, wherein the aerosol droplets have a mass median aerodynamic diameter of 3.0 μm to 3.3 μm, and the aerosol is characterized by a fine particle mass of at least 50% of the aerosol droplets, the fine particle mass representing the proportion of droplets having a diameter in the range of 1.0 to 5.0 μm.

[0169] In a preferred embodiment, the aerosol is characterized in that the aerosol droplets have a mass median aerodynamic diameter of 3.0 μm to 3.2 μm.

[0170] This applies to each of the aforementioned subtypes of inflammatory lung diseases as well as each of the aforementioned single inflammatory lung diseases.

[0171] Preferentially, the present application refers to the aforementioned embodiment in which 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt is used.

[0172] More preferably, the present application refers to the aforementioned embodiment in which crystalline polymorphic Form I, II or III of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt is used, as defined above by the respective d and / or 2θ values ​​determined by X-ray powder diagram.

[0173] Most preferably, the present application refers to the aforementioned embodiment in which the crystalline polymorphic Form I of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt is used, as defined above by the respective d and / or 2θ values ​​determined by X-ray powder diagram.

[0174] In another aspect of the present invention, the present application provides a method for treating a skin ulcer by administering to a subject a composition comprising 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts in the range of 0.01% by weight to 10% by weight. an aqueous solution ranging from 70% by weight to 99.99% by weight; Optionally, the term refers to an aerosol produced by a mesh nebulizer containing at least one pharmaceutically acceptable excipient in the range of 0% by weight to 20% by weight, the total of such percentages being 100%.

[0175] In particular, the present application refers to an aerosol produced by a vibrating mesh nebulizer containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts in the range of 0.01% by weight to 10% by weight, an aqueous solution in the range of 70% by weight to 99.99% by weight, and optionally at least one pharmaceutically acceptable excipient in the range of 0% by weight to 20% by weight, said percentages adding up to 100%, the aerosol droplets having a mass median aerodynamic diameter of 3.0 μm to 3.3 μm, and the aerosol characterized by a fine particle mass of at least 50% of the aerosol droplets, the fine particle mass representing the percentage of droplets having a diameter in the range of 1.0 to 5.0 μm.

[0176] In another aspect of the present invention, the present application provides a method for treating a skin ulcer by administering to a subject a composition comprising 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts in the range of 0.01% by weight to 10% by weight. an aqueous solution ranging from 70% by weight to 99.99% by weight; Optionally, reference is also made to a pharmaceutical composition for use in the prevention or treatment of inflammatory lung diseases comprising an aerosol generated by a nebulizer from an aqueous solution containing at least one pharmaceutically acceptable excipient in the range of 0% by weight to 20% by weight, the total of said percentages being 100%.

[0177] In another aspect of the present invention, the present application also provides a method for treating an inflammatory lung disease, comprising: a) providing an aerosol according to the invention by nebulization with a mesh nebulizer; and b) administering a therapeutically effective amount of said aerosol to a patient in need thereof through a mouthpiece for inhalation fitting on said mesh nebulizer by self-inhalation by the patient.

[0178] Formulations of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof may contain at least one pharmaceutically acceptable excipient.

[0179] The term "pharmaceutical excipient" refers to natural or synthetic compounds added to pharmaceutical formulations along with a pharmaceutically active agent. They serve to bulk the formulation, improve the formulation's desired pharmacokinetic properties or stability, and may be beneficial in the manufacturing process. Advantageous classes of excipients according to the present invention include colorants, buffers, preservatives, antioxidants, pH adjusters, solvents, tonicity agents, opacifiers, aromatic and flavoring substances.

[0180] Colorants are excipients that impart color to pharmaceutical formulations. These excipients can be food colorants. They can be adsorbed onto suitable adsorbents, such as clay or aluminum oxide. A further advantage of colorants is that they can make aqueous solutions spilled on the sprayer and / or mouthpiece visible for easier cleaning. The amount of colorant can vary from 0.01 to 10% by weight of the pharmaceutical composition, preferably from 0.05 to 6% by weight, more preferably from 0.1 to 4% by weight, and most preferably from 0.1 to 1% by weight.

[0181] Suitable pharmaceutical colorants are, for example, curcumin, riboflavin, riboflavin-5'-phosphate, tartrazine, alkanine, quinolion yellow WS, fast yellow AB, riboflavin-5'-sodium phosphate, yellow 2G, sunset yellow FCF, orange GGN, cochineal, carminic acid, citrus red 2, carmoisine, amaranth, ponceau 4R, ponceau SX, ponceau 6R, erythrosine, red 2G, allura red AC, indanthrene blue RS, patent blue V, indigo carmine, brilliant blue FCF, chlorophyll and chlorophyllin, copper complexes of chlorophyll and chlorophyllin, green S, fast green FCF, plain caramel, caustic sulfite caramel, ammonia caramel, AL, sulfite ammonia caramel, black PN, carbon black, vegetable carbon, brown FK, brown HT, alpha-carotene, beta-carotene, gamma-carotene, annatto, bixin, norbixin, paprika oleoleoresin, capsanthin, capsorubin, lycopene, beta-apo-8'-carotenal, ethyl ester of beta-apo-8'-carotenoic acid, flavoxanthin, lutein, cryptoxanthin, rubixanthin, violaxanthin, rhodoxanthin, canthaxanthin, zeaxanthin, citranaxanthin, astaxanthin, betanin, anthocyanin, saffron, calcium carbonate, titanium dioxide, iron oxide, iron hydroxide, aluminum, silver, gold, pigment rubin, tannin, orcein, iron gluconate, ferrous lactate.

[0182] Furthermore, buffers are preferred for liquid formulations, especially pharmaceutical liquid formulations. The terms buffer, buffer system, and buffer, especially for aqueous solutions, refer to the ability of a system to resist pH changes due to the addition of an acid or base or due to dilution with a solvent. Preferred buffer systems include formate, lactate, benzoate, oxalate, fumarate, aniline, acetate buffer, citrate buffer, glutamate buffer, phosphate buffer, succinate, pyridine, phthalate, histidine, MES (2-(N-morpholino)ethanesulfonic acid, maleic acid, cacodylate (dimethyl arsenate), carbonic acid, ADA (N-(2-acetamido)iminodiacetic acid), PIPES (4-piperazine-bis-ethanesulfonic acid), BIS-TRIS propane (1 ,3-bis[tris(hydroxymethyl)methylamino]propane), ethylenediamine, ACES (2-[(amino-2-oxoethyl)amino]ethanesulfonic acid), imidazole, MOPS (3-(N-morphino)-propanesulfonic acid, diethylmalonic acid, TES (2-[tris(hydroxymethyl)methyl]aminoethanesulfonic acid, HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), and pK values ​​of 3.8 to 7.7. a The buffer may be selected from the group including other buffers having the formula:

[0183] Carbonate buffers such as acetate buffers and dicarboxylic acid buffers such as fumarate, tartrate and phthalate, as well as tricarboxylic acid buffers such as citrate are preferred.

[0184] Another group of preferred buffers are inorganic buffers such as sulfate hydroxide, borate hydroxide, carbonate hydroxide, oxalate hydroxide, calcium hydroxide, and phosphate buffers. Another group of preferred buffers are nitrogen-containing buffers such as imidazole, diethylenediamine, and piperazine. Further preferred are sulfonic acid buffers such as TES, HEPES, ACES, PIPES, [(2-hydroxy-1,1-bis-(hydroxymethyl)ethyl)amino]-1-propanesulfonic acid (TAPS), 4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid (EEPS), 4-morpholino-propanesulfonic acid (MOPS), and N,N-bis-(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES). Another group of preferred buffers are glycine, glycylglycine, glycylglycylglycine, N,N-bis-(2-hydroxyethyl)glycine, and N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine (tricine). Also preferred are amino acid buffers such as glycine, alanine, valine, leucine, isoleucine, serine, threonine, phenylalanine, tyrosine, tryptophan, lysine, arginine, histidine, aspartic acid, glutamic acid, asparagine, glutamine, cysteine, methionine, proline, 4-hydroxyproline, N,N,N-trimethyllysine, 3-methylhistidine, 5-hydroxylysine, o-phosphoserine, gamma-carboxyglutamate, [epsilon]-N-acetyllysine, [omega]-N-methylarginine, citrulline, ornithine, and derivatives thereof.

[0185] Preservatives for liquid and / or solid dosage forms may be used as desired, including, but not limited to, sorbic acid, potassium sorbate, sodium sorbate, calcium sorbate, methylparaben, ethylparaben, methylethylparaben, propylparaben, benzoic acid, sodium benzoate, potassium benzoate, calcium benzoate, heptyl p-hydroxybenzoate, sodium methyl parahydroxybenzoate, sodium ethyl parahydroxybenzoate, sodium propyl parahydroxybenzoate, benzyl alcohol, benzalkonium chloride, phenylethyl alcohol, cresol, cetylpyridinium chloride, chlorobutanol, thiomer (sodium 2-(ethylmercurithio)benzoate), sulfur dioxide, sodium sulfite, sodium bisulfite, sodium metabisulfite, potassium metabisulfite, potassium sulfite, calcium sulfite. The antioxidant may be selected from the group comprising ammonium, calcium bisulfite, potassium bisulfite, biphenyl, orthophenylphenol, sodium orthophenylphenol, thiabendazole, nisin, natamycin, formic acid, sodium formate, calcium formate, hexamine, formaldehyde, dimethyl bicarbonate, potassium nitrite, sodium nitrite, sodium nitrate, potassium nitrate, acetic acid, potassium acetate, sodium acetate, sodium diacetate, calcium acetate, ammonium acetate, dehydroacetic acid, sodium dehydroacetate, lactic acid, propionic acid, sodium propionate, calcium propionate, potassium propionate, boric acid, sodium tetraborate, carbon dioxide, malic acid, fumaric acid, lysozyme, copper-(II)-sulfate, chlorine, chlorine dioxide, and other suitable substances or compositions known to those skilled in the art.

[0186] Suitable solvents may be selected from the group including, but not limited to, water, carbonated water, water for injection, water containing an isotonic agent, saline, isotonic saline, alcohols, particularly ethyl and n-butyl alcohol, and mixtures thereof.

[0187] Suitable isotonicity agents are, for example, pharmaceutically acceptable salts, in particular sodium chloride and potassium chloride, sugars such as glucose or lactose, sugar alcohols such as mannitol and sorbitol, citrates, phosphates, borates and mixtures thereof.

[0188] The addition of a sufficient amount of antioxidant is particularly preferred for liquid dosage forms. Suitable examples of antioxidants include sodium metabisulfite, alpha-tocopherol, ascorbic acid, maleic acid, sodium ascorbate, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, fumaric acid, or propyl gallate. Alpha-tocopherol and ascorbyl palmitate are preferred.

[0189] Suitable pH adjusting agents for liquid dosage forms are, for example, buffer substances such as sodium hydroxide, hydrochloric acid, sodium dihydrogen phosphate or disodium hydrogen phosphate.

[0190] Suitable aromatic and flavoring substances include all essential oils that can be used for this purpose. Generally, this term refers to volatile extracts from plants or plant parts that have their characteristic odor. They can be extracted from plants or plant parts by steam distillation.

[0191] Suitable examples are the essential oils of sage, clove, chamomile, anise, star anise, thyme, tea tree, peppermint, mint oil, menthol, cineole, borneol, gingerol, eucalyptus oil, mango, fig, lavender oil, chamomile flower, pine needles, cypress, orange, rosewood, plum, currant, cherry, birch leaf, cinnamon, lime, grapefruit, tangerine, juniper, valerian, lemon balm, lemongrass, palmarosa, cranberry, pomegranate, rosemary, ginger, pineapple, guava, echinacea, ivy leaf extract, blueberry, persimmon, melon, etc., their respective aromatics or mixtures thereof, as well as mixtures of menthol, peppermint and star anise oil or menthol and cherry flavors.

[0192] These aromatic or flavoring substances may be present in an amount ranging from 0.0001 to 10% by weight (particularly in the composition), preferably from 0.001 to 6% by weight, more preferably from 0.001 to 4% by weight, and most preferably from 0.01 to 1% by weight, relative to the total composition. Depending on the application or on a single occasion, it may be advantageous to use different amounts.

[0193] Opacifiers are substances that, if necessary, make a liquid dosage opaque. They must have a refractive index substantially different from that of the solvent, most often water. At the same time, they must be inert to the other components of the composition. Suitable examples include titanium dioxide, talc, calcium carbonate, behenic acid, cetyl alcohol, or mixtures thereof.

[0194] According to the present invention, all of the above-mentioned excipients and classes of excipients may be used without restriction, alone or in any conceivable combination thereof, as long as the use of the present invention is not hindered, toxic effects may occur or the legislation of the respective country is violated.

[0195] In another aspect of the invention, the present application also provides a method for producing an aerosol according to the invention, comprising: a) filling the spray chamber of a mesh nebulizer with 0.1 ml to 5 ml of an aqueous solution containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, and optionally at least one pharmaceutically acceptable excipient; b) starting vibration of the mesh of the mesh nebulizer at a frequency of 80 kHz to 200 kHz; and c) discharging the generated aerosol through the mesh of the mesh nebulizer on the side opposite the atomization chamber. Also mentioned is a method including:

[0196] The vibration frequency of a vibrating mesh nebulizer is typically in the range of 80 kHz to 200 kHz, preferably 90 kHz to 180 kHz, more preferably 100 kHz to 160 kHz, and most preferably 105 kHz to 130 kHz (see Chen, The Aerosol Society: DDL2019; Gardenshire et al. (2017) A Guide to Aerosol Delivery Devices for Respiratory Therapists, 4th ed.).

[0197] Therefore, the aforementioned method is also disclosed along with the vibration frequency range.

[0198] As seen in the aerosol analysis presented in Example 2, the method of the present invention has proven particularly effective in atomizing a high percentage of a pharmaceutically active agent from a provided aqueous solution, with relatively little loss of the pharmaceutically active agent during the atomization process.

[0199] The method according to the invention is therefore characterized in that at least 80% by weight, preferably at least 85% by weight, most preferably at least 90% by weight of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts contained in said aqueous solution is sprayed into the aerosol produced.

[0200] As seen in the aerosol analysis and respective experimental setup of Example 2, the method of the present invention proved particularly effective in nebulizing a high percentage of the pharmaceutically active agent from a provided aqueous solution within a short period of time. This is an important feature for patient compliance. A significant proportion of the patient population finds the inhalation process unpleasant, tedious, and exhausting. On the other hand, active patient cooperation is essential for effective and targeted inhalation applications. Therefore, it is desirable to administer a therapeutically sufficient amount within the shortest possible period of time. Surprisingly, it was shown that 95% of the substance provided in the aqueous solution could be nebulized within a 3-minute time span. This is an ideal period for high patient compliance.

[0201] Thus, the method according to the invention is characterized in that at least 80%, preferably at least 85%, and most preferably at least 90% of the aerosol produced is produced within 3 minutes after the start of nebulization with the mesh nebulizer.

[0202] While the pharmaceutically active agent is typically provided in a single-dose container for each nebulization procedure, the nebulizer and / or mouthpiece can be used for a certain period of time and must be replaced at regular intervals. Cleaning the nebulizer and mouthpiece is recommended by default after each nebulization. However, this does not allow for reasonable patient compliance. However, even after careful cleaning, some aerosol deposits always remain in the nebulization chamber, outlet, and / or mouthpiece. Because aerosols are generated from aqueous solutions, these deposits pose a risk of generating bacterial bioburden that can contaminate inhaled aerosols. Deposits may also block the pores in the mesh membrane of mesh nebulizers. Generally, nebulizers and / or mouthpieces must be replaced every week or two. Therefore, it is convenient to provide a combined product that combines the medication and nebulizer.

[0203] Therefore, in another aspect of the present invention, the present application also refers to a kit comprising a vibrating mesh nebulizer and a pharmaceutically acceptable container containing an aqueous solution containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, and optionally at least one pharmaceutically acceptable excipient.

[0204] In particular, the application refers to a kit comprising a vibrating mesh nebulizer and a pharmaceutically acceptable container having an aqueous solution containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, and optionally at least one pharmaceutically acceptable excipient, wherein the vibrating mesh nebulizer is capable of generating an aerosol from the aqueous solution, the aqueous solution being characterized by an aerosol droplet mass aerodynamic diameter of 3.0 μm to 3.3 μm and a fine particle mass of at least 50% of the aerosol droplets, the fine particle mass representing the proportion of droplets having a diameter in the range of 1.0 to 5.0 μm.

[0205] In another kit, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts is provided in two separate containers, one containing a solid form of the active agent and the other containing an aqueous solution. The final aqueous solution is freshly prepared by dissolving the active agent in the final solution. This final aqueous solution is then filled into the atomization chamber of a mesh nebulizer. These two containers can be completely separate containers, such as two vials, or a dual-chamber vial. To dissolve the active agent, for example, the membrane between the two chambers can be perforated to allow the contents of both chambers to mix.

[0206] Accordingly, the present application also discloses a kit comprising a vibrating mesh nebulizer, a first pharmaceutically acceptable container containing water for injection or saline, and a second pharmaceutically acceptable container containing a solid form of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, optionally with at least one pharmaceutically acceptable excipient contained in the first pharmaceutically acceptable container and / or the second pharmaceutically acceptable container.

[0207] In particular, the disclosure refers to a kit comprising a vibrating mesh nebulizer, a first pharmaceutically acceptable container containing water for injection or saline, and a second pharmaceutically acceptable container containing a solid form of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof, and optionally at least one pharmaceutically acceptable excipient is contained in the first pharmaceutically acceptable container and / or the second pharmaceutically acceptable container, wherein the vibrating mesh nebulizer is capable of generating an aerosol from a solution resulting from mixing of the contents of the first and second containers, the aerosol characterized by an aerosol droplet mass aerodynamic diameter of 3.0 μm to 3.3 μm and a fine particle mass of at least 50% of the aerosol droplets, the fine particle mass representing the proportion of droplets having a diameter in the range of 1.0 μm to 5.0 μm.

[0208] The aerosols produced by the method according to the invention are each administered and self-administered by means of a mouthpiece, which may optionally be further included in the aforementioned kit.

[0209] A common method for transferring the provided or final aqueous solution into the nebulization chamber of a mesh nebulizer is via a syringe equipped with a needle. The aqueous solution is first drawn up into the syringe and then injected into the nebulization chamber. Optionally, such a syringe and / or needle can be further included in the kit. Typical syringes that can be used include, but are not limited to, those made of polyethylene, polypropylene, or cyclic olefin copolymers, and typical gauges for stainless steel needles are in the range of 14-27. [Example]

[0210] 5-Amino-2,3-dihydro-1,4-phthalazinedione sodium salt Form I was provided by MetrioPharm Deutschland GmbH, Berlin, Germany. Salbutamol and ipratropium bromide were purchased from Merck KGaA, Darmstadt, Germany.

[0211] Example 1: 5-Amino-2,3-dihydro-1,4-phthalazinedione sodium salt Form I was tested in an isolated, ventilated, perfused mouse lung system challenged with cigarette smoke.

[0212] The isolated, ventilated, perfused mouse lung system (ILU) is an established model for studying the acute effects of various conditions and drugs on the lung parenchyma and vasculature. It is primarily used to examine the effects of hypoxia and evaluate the efficacy of potential drugs on hypoxic pulmonary vascular responses (see Weissmann et al. (2006) Proc Natl Acad Sci USA 103:19093-19098). Results from this experimental setting are considered indicative not only of the treatment of COPD, but also of all inflammatory disorders of the lower airways.

[0213] C57BL / 6J mice (n = 25, 5 per group; male / female, 3-6 months, 20-30 g; Charles River GmbH, Sulzfeld, Germany) were anesthetized with an intraperitoneal injection of ketamine (100 mg / kg body weight) and xylazine (20 mg / kg body weight) containing heparin (50 IE heparin / g body weight; Ratiopharm GmbH, Ulm, Germany) (Ceva Tiergesundheit GmbH, Düsseldorf, Germany). The lungs and heart were removed from the thoracic cavity and placed in an ILU system (see Figure 1A and Figure 1B). Lungs were ventilated in an isolated chamber using normoxic gas (21% O, 5% CO, 74% N; 150 breaths per minute with a PEEP (positive end-expiratory pressure) of 3 cm H2O) and perfused with modified Krebs-Henseleit buffer (120.0 mM NaCl, 4.3 mM KCl, 1.1 mM KH2PO4, 2.4 mM CaCl2, 1.3 mM MgCl2, 13.14 mM glucose, 0.25 mM hydroxyethyl starch 200,000 / 0.5, 25.0 mM NaHCO3, 800 mM L-arginine, adjusted to a constant pH range of 7.37–7.40; Serag-Wissner GmbH & Co. KG, Naira, Germany) at 37 °C. Lung weight, right and left ventricular pressures, and ventilation pressures were monitored and recorded throughout the experimental procedure. After 5–10 min, once the lungs had been adequately flushed and all parameters had stabilized, 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt was administered by adding 150 μl of the stock solution to 15 ml of perfusion buffer. This substance was administered 10 min before the first cigarette smoke application. While the lungs were perfused with buffer containing 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, cigarette smoke was administered via the trachea. Cigarette smoke was freshly prepared before each administration by burning one cigarette (University of Kentucky Research Cigarette 3R4F) for 1 min using normoxic gas at a flow rate of 1 L / min. The cigarette smoke was collected in a 1 L glass bottle containing 5 g of silica gel to remove moisture from the cigarette smoke. 50 ml of cigarette smoke was collected via a syringe and administered to the lungs via the trachea (Figure 1A) with deep breathing (periodic inflations of 3–4 s) over a 5 min period.The administration was performed manually, with careful monitoring of inspiratory pressure to avoid lung injury. The administration of cigarette smoke was repeated three times with a 1-hour break between each administration.

[0214] Five treatment groups (n=5 each) were investigated: A: Indoor air exposure B: Cigarette smoke + diluent (buffer solution) C: Cigarette smoke + 0.5 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt D: Cigarette smoke + 1 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt E: Cigarette smoke + 2 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt

[0215] 5-Amino-2,3-dihydro-1,4-phthalazinedione sodium salt Form I was dissolved in water for injection (vehicle) at the required concentration as above. A stock solution was prepared in water for injection. A further 1:100 dilution was made using modified Krebs-Henseleit buffer (see above). The stock solution was stored in appropriate aliquots at -70°C. The required amount of stock solution was thawed and the corresponding working solution was prepared for immediate use.

[0216] One hour after the third cigarette smoke application, the lungs were removed from the system and fixed for 2 hours at room temperature by inflating (through the trachea) with formalin solution at a pressure of 12-15 cm HO. Fixed lungs were then kept at +4°C in PBS (phosphate-buffered saline, see below) until further dehydration and paraffin embedding. Paraffin blocks were cut at 3 μm thickness, dried overnight at 37°C, and stained for 3-nitrotyrosine (3-NT).

[0217] Toxins and xenobiotics in tobacco smoke result in a dramatic increase in reactive oxygen species (ROS) and reactive nitrogen species (RNS). Oxidative and nitrosative stress correlate with the severity of inflammatory lung diseases. They increase the inflammatory response, cause an imbalance between proteolytic and antiproteolytic activity, increase the number of apoptotic cells, and decrease proliferation. These oxidants can overwhelm antioxidant defenses and initiate inflammation by various mechanisms (Foronjy and D'Armiento (2006) Clinical and Applied Immunology Reviews 6:53-72). The most potent RNS, peroxynitrite (ONOO), - ) is a product of nitric oxide (NO) and superoxide anion radical (O2 - ) (Szabo et al. (2007) Nat Rev Drug Discov 6:662-680). -Preferably, 3-NT attacks tyrosine residues in proteins to form the stable adduct 3-nitrotyrosine (Ricciardolo et al. (2004) Physiol Rev 84:731-765; Seimetz et al. (2011) Cell 147:293-305; Tsoumakidou et al. (2005) Chest 127:1911-1918). 3-NT levels in sputum proteins have been shown to be negatively correlated with FEV1 in COPD patients (Ricciardolo et al. (2004) Physiol Rev 84:731-765; Tsoumakidou et al. (2005) Chest 127:1911-1918). Nitrated tyrosine residues alter cell signaling, suggesting that 3-NT may not only be a marker of nitrosative stress but also have a functional relationship to the pathophysiology of inflammatory airway diseases (Davis et al. (2002); J Virol 76:8347-8359; Murata and Kawanishi (2004) Biochem Biophys Res Comm 316:123-128; Sugiura et al. (2004) Free Radic Res 38:49-57). 3-NT contributes to airway hyperresponsiveness and epithelial damage (Tsoumakidou et al. (2005) Chest 127:1911-1918) and has been proposed to play a major role in the development of airway remodeling (Ichinose et al. (2000) Am J Respir Crit Care Med 162:701-706).

[0218] Immunohistochemical staining for 3-nitrotyrosine was performed according to the following protocol: [Table 1]

[0219] Xylol was purchased from Carl Roth GmbH & Co. KG (Karlsruhe, Germany). Ethanol (96% and 99.6%) was purchased from Otto Fischar GmbH & Co. KG (Saarbrücken, Germany). Ethanol (70%) was purchased from SAV Liquid Production GmbH, Flintsbach am Inn, Germany. Hydrogen peroxide was purchased from Merck KGaA, Darmstadt, Germany. Methanol, bovine serum albumin (BSA), DAPI (4',6-diamidino-2-phenylidone), and anti-nitrotyrosine antibody (N0409; batch: 120M4825) were purchased from Sigma-Aldrich Co., Darmstadt, Germany. Rodent Decloacre's buffer (10x) and Warp Red Chromogen Kit were purchased from Biocare Medical, Pacheco, CA, USA. Tris wash buffer (TBS), CAT hematoxylin staining solution, and AP Polymer System (mouse / rabbit) were purchased from Zytomed Systems GmbH, Berlin, Germany. Dako Fluorescent Mounting Medium was purchased from Dako North America Inc., Villa Real Carpinteria, CA, USA. TruStain fcX (anti-mouse CD16 / 32; DR Fc block) was purchased from BioLegend Inc., San Diego, CA, USA. PBS (phosphate-buffered saline) was prepared using 8 g / L sodium chloride (Carl Roth GmbH+Co. KG, Karlsruhe, Germany), 0.2 g / L potassium chloride (Carl Roth GmbH+Co. KG, Karlsruhe, Germany), 1.42 g / L disodium hydrogen phosphate (Merck KGaA, Darmstadt, Germany), and 0.27 g / L potassium dihydrogen phosphate (Merck KGaA, Darmstadt, Germany).

[0220] Stained histological samples were analyzed blindly by light microscopy. 3-nitrotyrosine levels in the lung parenchyma were quantified as a percentage of the stained surface area. Quantification was performed at 200x magnification in 5-10 randomly selected fields, excluding large bronchi and blood vessels. For comparisons between groups, a one-way ANOVA statistical test with Bonferroni correction was performed. Differences of p<0.05 were considered statistically significant.

[0221] Cigarette smoke applied via the trachea resulted in a significant increase in 3-nitrotyrosine in the septum of exposed lungs (Figure 2B) compared with room air (Figure 2A). 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt was added to the perfusion buffer before cigarette smoke application and maintained throughout the experiment. Cigarette smoke-induced 3-nitrotyrosine formation could be almost completely abolished in lungs perfused with buffer containing 1 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt (Figure 2D) or 2 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt (Figure 2E), whereas the lowest 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt concentration (0.5 mM; Figure 2C) produced a moderate effect.

[0222] Quantification of staining: [Table 2]

[0223] The results are shown as a bar graph in Figure 3. Values ​​(mean ± SEM) indicate the percentage of stained surface in the histological samples evaluated (5 mice per group; 5–6 evaluated histological samples per mouse).

[0224] From this experiment, it can be concluded that pretreatment with 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt prevents cigarette smoke-induced 3-nitrotyrosine formation in the lung parenchyma of the ILU model.

[0225] This suggests that 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt has a protective effect against acute cigarette smoke-induced lung injury. Therefore, these results can be considered predictive of the beneficial effects of 5-amino-2,3-dihydro-1,4-phthalazinedione and its pharmaceutically acceptable salts in the inhalation prophylaxis or treatment of all inflammatory lung diseases.

[0226] Example 2: The particle size distribution (FPM) and mass median aerodynamic diameter (MMAD) of aerosols generated from aqueous solutions containing 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt were analyzed.

[0227] Polymorphic Form I of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt was dissolved in saline (double-distilled water containing 0.9% NaCl) to a final concentration of 20 mg / ml of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt.

[0228] The experimental procedure was performed in accordance with European Pharmacopoeia 2.9.44 (Preparation for Nebulization: Characterization). The proportion of each fraction of nebulized particles was determined using a cascade impactor (Next Generation Impactor (NGI), Copley Scientific Ltd., Nottingham, UK). The cascade impactor was cooled to 4°C for the measurements.

[0229] NGI includes the following features: 1) Designed and accepted by the pharmaceutical and biotechnology industries for inhalation testing; 2) Meets and exceeds all European Pharmacopoeia and United States Pharmacopoeia specifications; 3) particle size range of 0.24–11.7 μm (dependent on flow rate); 4) Five of the seven stages have a cutoff of 0.54 to 6.12 μm at flow rates of 30 to 100 l / min; 5) Calibrated flow range of 30~100L / min; 6) Further calibration at 15 l / min for nebulizer applications; 7) Submit full stage measurement report (system suitability); 8) Low interstage wall losses (mass balance) for good drug recovery; 9) Conductive and not affected by static electricity; The NGI cascade impactor itself comprises three main parts: a) A cup tray containing eight collection cups used to collect samples prior to analysis b) A bottom frame used to support the cup tray c) The lid includes an interstage passageway and a seal that holds the nozzle in place.

[0230] The 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt solution was nebulized using a vibrating mesh nebulizer (M-neb-dose+, NEBU-TEC, Eisenfeld, Germany), and the aerosol thus generated was delivered to a cascade impactor via a mouthpiece (SK-211, NEBU-TEC, Eisenfeld, Germany).

[0231] The nebulization chamber (yellow stamp, 250 μl liquid output) was filled. The puff profile configured with the breath simulator was selected to ensure accurate simulation of inhalation by the nebulizer. The corresponding time was recorded.

[0232] Quantification of nebulized 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt per cascade was performed by HPLC after extraction from the cascade pans (stages 1–8) with reference to an external standard calibration curve (range 0–200 μg / ml; correlation coefficient: 0.9999).

[0233] The single values ​​obtained for each cascade were added together. No residual amounts of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt were found in the mouthpiece. MMAD, FPF, and FPM were calculated from the obtained values. [Table 3]

[0234] The cumulative particle size distribution is shown in Figure 4A, and the relative particle size distribution is shown in Figure 4B.

[0235] The aerosol can be further characterized as follows: -The emitted dose at the 4.7 mg mouthpiece corresponds to 95% of the nominal value -Aerosol release time was 3 minutes (equivalent to 30 strokes / breaths). - The ratio of particles with MMAD<1 μm to particles with MMAD>1 μm is 5:95.

[0236] Example 3: For comparison, the particle size distribution (FPF and FPM) and mass median aerodynamic diameter (MMAD) of aerosols generated from saline containing salbutamol sulfate were analyzed (not part of the present invention). The procedure was the same as that described in Example 2.

[0237] Salbutamol (albuterol) is a short-term bronchodilator. It consists of a racemic mixture of the pharmacologically active (R)-(-)-enantiomer and the metabolically active (S)-(+)-enantiomer, which act as a beta-2 receptor agonist. Activation of beta-2 receptors results in the conversion of ATP to cyclic AMP (cAMP) via adenylyl cyclase, inhibiting myosin phosphorylation and Ca2+ upregulation. 2+ The increase in cAMP also inhibits inflammatory cells in the airways from releasing inflammatory mediators and cytokines. Salbutamol also inhibits Ca 2+ Channel and K +It increases the conductance of the channel, resulting in hyperpolarization and relaxation of bronchial smooth muscle.

[0238] Salbutamol is one of the most widely used substances for acute asthma attacks. It is often administered via metered-dose inhalers. Therefore, saline solutions of salbutamol sulfate were tested to determine particle size distribution. A standard dose of 180 μg (90 μg / spray, 2 sprays per inhalation session) of salbutamol sulfate (albutol sulfate) was dissolved in saline.

[0239] The single values ​​obtained for each cascade were added. No residual amounts of salbutamol sulfate were found in the mouthpiece. MMAD, FPF, and FPM were calculated from the obtained values. [Table 4]

[0240] The cumulative particle size distribution is shown in Figure 5A, and the relative particle size distribution is shown in Figure 5B.

[0241] The aerosol can be further characterized as follows: -The emitted dose at the 166 μg mouthpiece corresponds to 92% of the nominal value -Aerosol release time was 3 minutes (equivalent to 30 strokes / breaths). - The ratio of particles with MMAD<1 μm to particles with MMAD>1 μm is 4:96.

[0242] In comparison, 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt exhibits a significantly smaller MMAD than salbutamol sulfate. The FPF is slightly higher for 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, while the FPM is in a similar range.

[0243] Example 4: For comparison, the particle size distribution (FPF and FPM) and mass median aerodynamic diameter (MMAD) of aerosols generated from saline containing ipratropium bromide were analyzed (not part of this invention). The procedure was the same as that described in Example 2.

[0244] Ipratropium bromide is a pharmaceutically active drug used to treat COPD and cardiac arrhythmias. It acts as a competitive antagonist of muscarinic acetylcholine receptors, M3 receptors, in the smooth muscle of the bronchial and pulmonary vascular epithelium. M3 receptors are G q coupled to an increase in inositol triphosphate (IP3), followed by Ca from intracellular calcium stores. 2+ Blockade of M3 receptors therefore leads to bronchodilation.

[0245] Ipratropium bromide is often administered via nebulizer and nasal spray. Therefore, a saline solution of ipratropium bromide was tested to determine particle size distribution. A standard dose of 18 μg / spray application (Atrovent) was dissolved in saline.

[0246] The single values ​​obtained for each cascade were added. No residual ipratropium bromide was found in the mouthpiece. MMAD, FPF, and FPM were calculated from the obtained values. [Table 5]

[0247] The cumulative particle size distribution is shown in Figure 6A, and the relative particle size distribution is shown in Figure 6B.

[0248] The aerosol can be further characterized as follows: The emitted dose at the 15 μg mouthpiece corresponds to 86% of the nominal value -Aerosol release time was 3 minutes (equivalent to 30 strokes / breaths). - The ratio of particles with MMAD<1 μm to particles with MMAD>1 μm is 4:96.

[0249] In comparison, 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt exhibits a significantly smaller MMAD than ipratropium bromide. The FPF is slightly higher and the FPM is slightly lower for 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt. [Brief explanation of the drawings]

[0250] [Figure 1] A: Schematic diagram of the experimental setup of Example 1. 1 - Cigarette smoke 2 - Ventilator 3 - Trachea 4 - Lungs 5 ​​- Heart 6 - Reservoir 7 - Aqueous solution of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt 8 - Roller pump B: Photograph of the experimental setup of Example 1. [Figure 2] Immunohistochemical staining of a representative sample from Example 1. Left panel: 200x magnification. Right panel: 400x magnification, more detailed enlargement than the left panel. A: Room air. B: Cigarette smoke + diluent (buffer). C: Cigarette smoke + 0.5mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt. D: Cigarette smoke + 1mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt. E: Cigarette smoke + 2mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt. In the right panel, the inflamed area is highlighted. [Figure 3] Statistical evaluation of immunohistochemical staining of samples from Example 1. The percentage of stained surface area corresponds to the inflammation grade (n=5; mean±SEM). Bars with asterisks indicate highly significant differences between both groups (p<0.001). A: Room air B: Cigarette smoke + diluent (buffer) C: Cigarette smoke + 0.5 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt D: Cigarette smoke + 1 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt E: Cigarette smoke + 2 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt [Figure 4]Particle size distribution (μm) of the aerosol produced in Example 2, determined as MMAD. A: Cumulative MMAD B: Relative MMAD [Figure 5] Particle size distribution (μm) of the aerosol produced in Example 3, determined as MMAD. A: Cumulative MMAD B: Relative MMAD [Figure 6] Particle size distribution (μm) of the aerosol produced in Example 4, determined as MMAD. A: Cumulative MMAD B: Relative MMAD

[0251] List of abbreviations: [Table 6] TIFF0007766035000007.tif107166

Claims

1. A composition for use by inhalation administration using an aerosol, which contains 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts as an active ingredient, the aerosol droplets have a mass median aerodynamic diameter of 3.0 μm to 3.3 μm; the aerosol is characterized in that a fine particle mass is at least 50% of the droplets of the aerosol, the fine particle mass representing a proportion of the droplets having a diameter in the range of 1.0 to 5.0 μm; A preventive or therapeutic agent for inflammatory lung diseases.

2. at least 50% of the droplets of the aerosol have a mass aerodynamic particle diameter of 1.0 μm to 3.2 μm; The preventive or therapeutic agent for inflammatory lung diseases according to claim 1.

3. The pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt. The preventive or therapeutic agent for inflammatory lung diseases according to claim 1 or 2.

4. 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt has the X-ray powder diagram: For Form I, d values: 13.5; 6.9; 5.2; 4.6; 3.9; 3.5; 3.4; 3.3; 3.1; 3.0 and / or 2θ values: 6.5; 12.7; 16.9; 19.3; 22.8; 25.8; 26.6; 27.2; 28.7; 30.3, For Form II, d values: 12.9; 7.9; 7.1; 6.5; 5.3; 4.0; 3.7; 3.6; 3.3; 3.2 and / or 2θ values: 6.8; 11.2; 12.5; 13.7; 16.7; 22.4; 24.3; 24.9; 27.2; 27.8, and For Form III, d values: 13.131; 7.987; 7.186; 6.566; 6.512; 5.372; 3.994; 3.662; 3.406; 3.288; 3.283; 3.222; 3.215; 3.127; 2.889 and / or 2θ values: 6.73; 11.07; 12.31; 13.48; 13.59; 16.49; 22.24; 24.29; 26.14; 27.10; 27.14; 27.67; 27.72; 28.52; 30.93 The compound is provided as one of crystalline anhydrous polymorphic forms I, II or III characterized by a crystallographic value determined by The preventive or therapeutic agent for inflammatory lung diseases according to claim 3.

5. The inflammatory lung disease is selected from the group comprising inflammation of the lower respiratory tract due to bacterial, viral, fungal or parasitic infection, chronic lower respiratory tract disease, lung disease due to external agents, respiratory diseases mainly affecting the interstitium, suppurative and / or necrotic conditions of the lower respiratory tract, pleural diseases, postoperative or related lower respiratory tract diseases, perinatal-specific lung diseases, trauma and injury of the lower respiratory tract and / or thorax, and malignant neoplasms of the lower respiratory tract, The preventive or therapeutic agent for inflammatory lung diseases according to any one of claims 1 to 4.

6. The inflammatory lung disease is selected from the group consisting of chronic obstructive pulmonary disease, asthma, pulmonary sarcoidosis, cystic fibrosis, bronchiectasis, adult respiratory distress syndrome, pulmonary fibrosis, beryllium pulmonary disease, chronic lung allograft dysfunction, pulmonary edema, pulmonary ischemia-reperfusion injury, and primary graft dysfunction after lung transplantation. The preventive or therapeutic agent for inflammatory lung diseases according to any one of claims 1 to 5.

7. 1. An aerosol comprising 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts in the range of 0.01% to 10% by weight, an aqueous solution in the range of 70% by weight to 99.99% by weight, and optionally at least one pharmaceutically acceptable excipient in the range of 0% by weight to 20% by weight, The percentages add up to 100%, The aerosol droplets have a mass median aerodynamic diameter of 3.0 μm to 3.3 μm, and the aerosol is characterized in that the fine particle mass exhibits a proportion of droplets having a diameter in the range of 1.0 to 5.0 μm, with at least 50% of the droplets of the aerosol being at least 50% at most. aerosol.

8. a) filling the atomizing chamber of a mesh nebulizer with 0.1 ml to 5 ml of an aqueous solution containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, and optionally at least one pharmaceutically acceptable excipient; b) initiating vibration of the mesh of the mesh nebulizer at a frequency of 80 kHz to 200 kHz; and c) discharging the generated aerosol through the mesh nebulizer on the side opposite the nebulization chamber from the mesh. Including, 8. A method for producing the aerosol of claim 7.

9. At least 90% by weight of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts contained in the aqueous solution is sprayed into the generated aerosol. The method of claim 8.

10. At least 80% of the generated aerosol is generated within 3 minutes after initiation of nebulization in the mesh nebulizer.

10. The method according to claim 8 or 9.

11. 1. A kit comprising a vibrating mesh nebulizer and a pharmaceutically acceptable container having an aqueous solution containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, and optionally at least one pharmaceutically acceptable excipient, the vibrating mesh nebulizer is capable of generating an aerosol from the aqueous solution, the aerosol having droplets with a mass median aerodynamic diameter of 3.0 μm to 3.3 μm; and a fine particle mass of at least 50% of the droplets of the aerosol, the fine particle mass representing a proportion of droplets having a diameter in the range of 1.0 to 5.0 μm; kit.

12. 1. A kit comprising a vibrating mesh nebulizer, a first pharmaceutically acceptable container containing water for injection or saline, and a second pharmaceutically acceptable container containing a solid form of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof, Optionally, at least one pharmaceutically acceptable excipient is contained in said first pharmaceutically acceptable container and / or said second pharmaceutically acceptable container; the vibrating mesh nebulizer is capable of generating an aerosol from a solution resulting from mixing the contents of the first container and the second container; the aerosol has a mass median aerodynamic diameter of the droplets of the aerosol of 3.0 μm to 3.3 μm; and a fine particle mass of at least 50% of the droplets of said aerosol; The fine particle mass indicates the proportion of droplets having a diameter in the range of 1.0 μm to 5.0 μm. kit.

13. Further comprising a mouthpiece for inhalation fitting to the mesh nebulizer.

13. The kit according to claim 11 or 12.

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