Compounds for the treatment of coronavirus infections

Abstract

This application relates to the use of phthalazinediones in the prevention or treatment of coronavirus infections. Pharmaceutical compositions, combinations and advantageous formulation techniques are disclosed.

Description

[Technical Field]

[0001] This application relates to the use of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for the prevention or treatment of coronavirus infection. More specifically, this application relates to the sodium salt of 5-amino-2,3-dihydro-1,4-phthalazinedione for the prevention or treatment of coronavirus infection. Pharmaceutical compositions, combinations, and advantageous dosage forms are disclosed. [Background technology]

[0002] As a result of ecological, climatic and demographic changes, so-called 'emerging' viruses are increasingly being transmitted from natural animal hosts to humans, and with the acceleration of globalization, they are at risk of causing pandemics.

[0003] Emerging viruses can cause acute and often life-threatening illnesses. Coronaviruses are notorious for such contagion. Examples include severe acute respiratory syndrome coronavirus 1 (SARS-CoV-1), Middle East respiratory syndrome-related coronavirus (MERS-CoV), and most recently, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; COVID-19) in Wuhan, China. As of May 31, 2021, the Johns Hopkins University Coronavirus Resource Center reported a total of over 170 million SARS-CoV-2 cases and approximately 3.5 million deaths worldwide. The incubation period for SARS-CoV-2 ranges from two days to two weeks, and in some cases, up to one month. The disease resulting from SARS-CoV-2 infection is called COVID-19.

[0004] Typical symptoms of COVID-19 are fever, cough, and shortness of breath. However, the infection can also cause severe lung damage, which can rapidly develop into progressive lung dysfunction, particularly with regard to the lung's ability to take in oxygen. This is usually associated with the failure of other organs.

[0005] This acute lung injury (ALI) state is associated with widespread pulmonary inflammation and intraalveolar fluid accumulation. It is characterized by diffuse pulmonary microvascular injury resulting in increased permeability and consequent noncardiogenic pulmonary edema. Consequently, this leads to pathologically low oxygen levels in the lungs. Other common conditions associated with COVID-19 patients in ICU care are pulmonary embolism, thrombosis, venous thromboembolism, and cerebral ischemia.

[0006] Coronaviruses spread primarily through respiratory droplets from coughs and sneezes during close contact. In contrast to SARS-CoV and MERS-CoV, SARS-CoV-2 can be transmitted from person to person during the incubation period, although infected patients do not show symptoms of illness for some time. Furthermore, SARS-CoV-2 replicates early in the throat. In contrast, the receptors for SARS-CoV and MERS-CoV are located deep in the lungs. Therefore, SARS-CoV-2 is much more easily transmitted from person to person compared to SARS-CoV and MERS-CoV, significantly increasing its infection rate.

[0007] In general, coronaviruses (family Coronaviridae, group Coronaviridae) form a large and relatively diverse group of enveloped, positive-strand RNA viruses that can cause various types of diarrhea and respiratory diseases in humans and animals. They have a very narrow host range and replicate very poorly in cell culture. However, we have successfully established a cell culture system for SARS-CoV-2.

[0008] Determination of the nucleotide sequence of SARS-CoV-2 revealed a genome of approximately 29.8 kbp consisting of 14 open reading frames. Furthermore, this virus was phylogenetically closely related to SARS-CoV (nucleotide similarity: 89.1%) (Reference: Wu et al. (2020) Nature, Epub ahead of print). Like other coronaviruses, SARS-CoV-2 enters cells through endocytosis and membrane fusion. The virus is released from cells via the secretory pathway. The natural host of the virus is unknown.

[0009] To date, no specific therapeutic options have been established for the treatment of SARS-CoV-2 infection, respectively COVID-19. The antiviral drugs remdesivir, avifavir, and favipiravir have shown some success. A nasal spray containing nanoantibodies against the SARS-CoV-2 spike protein is a promising development (AeroNabs). In patients with severe COVID-19, administration of the glucocorticoid dexamethasone has been shown to be effective.

[0010] Therefore, there is a strong medical need for effective drug therapies for patients infected with SARS-CoV-2 and similar coronaviruses and to contain the current outbreak of this virus. Ideally, such drug therapies would at least be a treatment option for future coronavirus outbreaks.

[0011] Surprisingly, this problem is solved by administering 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 thereof.

[0012] Accordingly, the present application discloses 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of coronavirus infection. DETAILED DESCRIPTION OF THE INVENTION

[0013] 5-Amino-2,3-dihydro-1,4-phthalazinedione (luminol) belongs to the pharmaceutical class of phthalazinediones. 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 applied in diagnostic assays and forensics as a detection tool, for example, in blood spot tracing. In medicine, 5-amino-2,3-dihydro-1,4-phthalazinedione has been developed in the form of its sodium salt. In some countries, it has been 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, inflammation such as prostatitis, endometriosis, throat inflammation, bronchial asthma, pneumonia, periodontitis, pyelonephritis, and autoimmune diseases such as Crohn's disease, ulcerative colitis, lupus erythematosus, and scleroderma. 5-Amino-2,3-dihydro-1,4-phthalazinedione could effectively prevent cytokine storms caused by excessive immune responses. Furthermore, the scientific and patent literature still contains a long list of therapeutic indications for which 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt has been tested or has been suggested to be of beneficial use (see, inter alia, WO 2004 / 041169, WO 2007 / 018546, WO 2012 / 127441, WO 2016 / 096143, WO 2017 / 202496, WO 2018 / 082814).

[0014] 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.

[0015] To ensure better solubility and bioavailability, 5-amino-2,3-dihydro-1,4-phthalazinedione is used as a pharmaceutically acceptable salt. Sodium, potassium, and lithium salts have been described for therapeutic applications (see WO 2010 / 082858). The crystal structures of lithium, sodium, potassium, rubidium, and cesium salts were 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.

[0016] Accordingly, the present application discloses 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of coronavirus infection.

[0017] 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 reducing or inhibiting the replication of coronavirus in humans.

[0018] 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 reducing coronavirus viral load in humans.

[0019] 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 the prevention or treatment of COVID-19 in humans.

[0020] 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 the prevention or treatment of a coronavirus infection in a human, wherein the human is asymptomatic.

[0021] 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 the prevention or treatment of coronavirus infection in a human, wherein the human exhibits mild coronavirus infection-related symptoms and does not require hospitalization.

[0022] 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 treating a coronavirus infection in a human, wherein the human exhibits severe coronavirus infection-related symptoms and requires hospitalization.

[0023] In another aspect, the present application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in treating a coronavirus infection in a human, wherein the human exhibits severe coronavirus infection-associated symptoms and suffers from acute lung injury.

[0024] In particular, the present application discloses 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of coronavirus infection, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt.

[0025] 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 reducing or inhibiting coronavirus replication in humans, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt.

[0026] 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 reducing the viral load of coronavirus in humans, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt.

[0027] 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 the prevention or treatment of COVID-19 in humans, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt.

[0028] 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 the prevention or treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human is asymptomatic.

[0029] 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 the prevention or treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human shows mild coronavirus infection-related symptoms and does not require hospitalization.

[0030] 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 the treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and the human is exhibiting severe coronavirus infection-related symptoms and requiring hospitalization.

[0031] 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 treating a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and the human is exhibiting severe coronavirus infection-related symptoms and suffering from acute lung injury.

[0032] 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 or one of its pharmaceutically acceptable salts may form a complex with a suitable ligand. Therefore, this patent application also refers to such complexes. Within the scope of this disclosure, all hydrates and solvates are intended to be encompassed by the term "5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts."

[0033] Anhydrous formulations are often preferred to ensure reproducible, standardized API production and provide improved stability characteristics of the active drug. The anhydrous forms of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt are described as 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.

[0034] 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 2Theta values: 6.5; 12.7; 16.9; 19.3; 22.8; 25.8; 26.6; 27.2; 28.7; 30.3.

[0035] Form II is characterized by the following values: d-values: 12.9; 7.9; 7.1; 6.5; 5.3; 4.0; 3.7; 3.6; 3.3; 3.2 and / or 2Theta values: 6.8; 11.2; 12.5; 13.7; 16.7; 22.4; 24.3; 24.9; 27.2; 27.8.

[0036] 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 2 Theta 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.

[0037] The use of the anhydrous form I of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt is preferred.

[0038] In a mouse COPD model, ex vivo lungs were exposed to cigarette smoke. Application of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt form I resulted in nearly complete reduction of 3-nitrotyrosine. Therefore, it is hypothesized that 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt may protect lungs from oxidative and nitrosative stress, which are key players in the pathophysiology of COPD and other inflammatory lung diseases (https: / / copdnewstoday.com / 2020 / 04 / 16 / mp1032-may-protect-lungs-from-oxidative-stress-inhibit-biomarker-3-nitrotyrosine-preclinical-study / as of 2021-03-03).

[0039] 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.

[0040] WO 2017 / 140430 also discloses that 5-amino-2,3-dihydro-1,4-phthalazinedione has great potential in the immunomodulatory treatment of inflammatory and autoimmune diseases. Crystalline Form II is particularly useful for the treatment of inflammatory and autoimmune respiratory diseases, such as upper and lower respiratory tract infections.

[0041] Therefore, the present patent application also refers to the use according to the invention of all crystalline forms and polymorphs of 5-amino-2,3-dihydro-1,4-phthalazinedione and its pharmaceutically acceptable salts. The use of Form II of 5-amino-2,3-dihydro-1,4-phthalazinedione is preferred.

[0042] 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.

[0043] EP 0 531 370 A1 discloses the use of PARP (poly(ADP-ribose) polymerase) inhibitors, such as luminol, for the treatment of viral infections in which viral DNA is integrated into host chromosomes during the replication cycle. The virus is preferably a retrovirus, such as HIV. However, coronaviruses are positive-strand RNA viruses and therefore cannot integrate into human DNA. Therefore, EP 0 531 370 A1 does not suggest the use of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for the treatment of coronavirus infections.

[0044] Tautomerism refers to the rapid internal transformation of organic compounds in which a hydrogen atom or proton formally moves 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.

[0045] Isomers are molecules with the same chemical formula but different chemical structures. They can be distinguished into constitutional isomers (where atoms or functional groups are exchanged) 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) and 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 can occur in phthalazinedione derivatives. 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.

[0046] 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, this patent application also refers to such isotopically enriched forms of the compounds of the invention.

[0047] 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 into a metabolically active form in the body. This conversion can occur systemically or locally. Therefore, this patent application also refers to prodrugs of the compounds of the present invention.

[0048] As used throughout this application, the term "5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts" is intended to encompass 5-amino-2,3-dihydro-1,4-phthalazinedione, i.e., all the aforementioned molecular variants 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.

[0049] Unless otherwise defined, all technical or scientific terms used herein have the meanings ascribed to them by one of ordinary skill in the relevant art.

[0050] 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.

[0051] Coronavirus infections that can be treated with 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts include, among others, infections caused by highly pathogenic SARS-CoV-1, MERS-CoV, and SARS-CoV-2. However, infections caused by less pathogenic coronaviruses, such as those listed below, can also be treated in this manner. The term "coronavirus" or "coronavirus" primarily refers to the Orthocoronavirus subfamily, which is subdivided into the genera Alphacoronavirus, Betacoronavirus, Gammacoronavirus, and Deltacoronavirus. Alphacoronaviruses include the subgenera Coracovirus (species: bat coronavirus CDPHE15), Decavirus (bat coronavirus HKU10, horseshoe bat alphacoronavirus HuB-2013), Dubinacovirus (human coronavirus 229E), Lucacovirus (Routhillon Rn rat coronavirus), Minakovirus (ferret coronavirus, mink coronavirus 1), Minunacovirus (minioptera bat coronavirus 1, minioptera bat coronavirus HKU8), Myotakovirus (bigfoot bat alphacoronavirus Sax-2011), Nylactovirus (mountain long-eared bat alphacoronavirus SC-2013), Pedakovirus (porcine epidemic diarrhea virus, Scotophilus bat coronavirus 512), Rhynacovirus (horseshoe bat coronavirus HKU2), Setracovirus (human coronavirus NL63, NL63-related bat coronavirus strain BtKYNM63-9b), and Tegacovirus (alphacoronavirus type 1 species).Betacoronaviruses include the subgenus Enbecovirus (Betacoronavirus 1 (subspecies: Human coronavirus OC43), Chinese rat coronavirus HKU24, Human coronavirus HKU1, Murine coronavirus type species), Hibecovirus (Bat Hp-Betacoronavirus Chou Qiang 2013), Merbecovirus (Hedgehog coronavirus 1, MERS-CoV), Pipistrels bat coronavirus HKU5, Tyronycteris bat coronavirus HKU4), Nobecovirus (Rouset's flying fox coronavirus GCCDC1, Rouset's flying fox coronavirus HKU9), and Sarbecovirus (Severe acute respiratory syndrome-associated coronavirus (subspecies: SARS-CoV-1, SARS-CoV-2)). Gammacoronaviruses include the subgenus Sars-CoV (Severe acute respiratory syndrome-associated coronavirus (subspecies: SARS-CoV-1, SARS-CoV-2)). The Deltacoronaviruses include the subgenera Andechovirus (Wild Whale Coronavirus SW1) and Igakovirus (Avian Coronavirus type species). The Deltacoronaviruses include the subgenera Andechovirus (Wild Whale Coronavirus HKU20), Burdecovirus (Burboulder Coronavirus HKU11 type species, Porcine Coronavirus HKU15, Munia Coronavirus HKU13, White-eye Coronavirus HKU16), Herdecovirus (Night Heron Coronavirus HKU19), and Murdecovirus (Bank Coronavirus HKU21).

[0052] To date, the coronaviruses that are pathogenic in humans are SARS-CoV, SARS-CoV-2, MERS-CoV, and HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E, the last four of which cause only relatively mild symptoms (see Andersen et al.: The Proximal Origin of SARS-CoV-2, on virologica.org, as of February 17, 2020).

[0053] Thus, the present application particularly 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 a coronavirus infection, said coronavirus infection being selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43 and HCoV-229E infections.

[0054] In another aspect, the application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in reducing or inhibiting the replication of a coronavirus in a human, wherein said coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43 and HCoV-229E infection.

[0055] In another aspect, the present application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in reducing the viral load of a coronavirus in a human, wherein said coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43 and HCoV-229E infection.

[0056] In another aspect, the application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of a coronavirus infection in a human, wherein the human is asymptomatic and the coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43 and HCoV-229E infection.

[0057] In another aspect, the application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of a coronavirus infection in a human, wherein the human exhibits mild coronavirus infection-associated symptoms and does not require hospitalization, and the coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E infection.

[0058] In another aspect, the application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the treatment of a coronavirus infection in a human, wherein the human exhibits severe coronavirus infection-related symptoms and requires hospitalization, and the coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E infection.

[0059] In another aspect, the present application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the treatment of a coronavirus infection in a human, wherein the human is exhibiting severe coronavirus infection-associated symptoms and suffering from acute lung injury, and the coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E infection.

[0060] 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 reducing or inhibiting the replication of coronavirus in humans, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and said coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43 and HCoV-229E infection.

[0061] 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 reducing the viral load of a coronavirus in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein said coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E infection.

[0062] 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 the prevention or treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and the human is asymptomatic, and the coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E infection.

[0063] 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 the prevention or treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human exhibits mild coronavirus infection-associated symptoms and does not require hospitalization, and wherein the coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E infection.

[0064] 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 the treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human exhibits severe coronavirus infection-related symptoms and requires hospitalization, and the coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E infection.

[0065] 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 treating a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human is exhibiting severe coronavirus infection-associated symptoms and suffering from acute lung injury, and the coronavirus infection is selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1, HCoV-NL-63, HCoV-OC43, and HCoV-229E infection.

[0066] Therefore, the present application particularly relates to a 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of a coronavirus infection, said coronavirus infection being a SARS-CoV-2 infection.

[0067] In another aspect, the present application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in reducing or inhibiting the replication of coronavirus in humans, wherein said coronavirus infection is SARS-CoV-2 infection.

[0068] In another aspect, the present application refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in reducing the viral load of a coronavirus in a human, wherein the coronavirus infection is a SARS-CoV-2 infection.

[0069] In another aspect, the present application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of a coronavirus infection in a human, wherein the human is asymptomatic and the coronavirus infection is a SARS-CoV-2 infection.

[0070] In another aspect, 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 a coronavirus infection in a human, wherein the human exhibits mild coronavirus infection-related symptoms and does not require hospitalization, and the coronavirus infection is a SARS-CoV-2 infection.

[0071] In another aspect, the application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the treatment of a coronavirus infection in a human, wherein the human exhibits severe coronavirus infection-related symptoms and requires hospitalization, and the coronavirus infection is a SARS-CoV-2 infection.

[0072] In another aspect, the present application refers to one of 5-amino-2,3-dihydro-1,4-phthalazinedione or a pharmaceutically acceptable salt thereof for use in the treatment of a coronavirus infection in a human, wherein the human exhibits severe coronavirus infection-associated symptoms and suffers from acute lung injury, and the coronavirus infection is a SARS-CoV-2 infection.

[0073] 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 reducing or inhibiting coronavirus replication in humans, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and said coronavirus infection is SARS-CoV-2 infection.

[0074] 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 reducing the viral load of coronavirus in humans, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein said coronavirus infection is SARS-CoV-2 infection.

[0075] 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 the prevention or treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human is asymptomatic, and the coronavirus infection is a SARS-CoV-2 infection.

[0076] 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 the prevention or treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human shows mild coronavirus infection-related symptoms and does not require hospitalization, and wherein the coronavirus infection is a SARS-CoV-2 infection.

[0077] 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 the treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human exhibits severe coronavirus infection-related symptoms and requires hospitalization, and wherein the coronavirus infection is a SARS-CoV-2 infection.

[0078] 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 the treatment of a coronavirus infection in a human, wherein the pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt, and wherein the human is exhibiting severe coronavirus infection-associated symptoms and suffering from acute lung injury, and wherein the coronavirus infection is a SARS-CoV-2 infection.

[0079] More preferred is 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of coronavirus infection, wherein said coronavirus infection is SARS-CoV-2.

[0080] Most preferred is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt for use in the prevention or treatment of coronavirus infection, wherein said coronavirus infection is SARS-CoV-2.

[0081] Although the other above-mentioned animal coronaviruses have not yet been transferred to humans (zoonotic diseases), this may occur in the future with unpredictable pathology. Therefore, the scope of the present application also relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for use in the prevention or treatment of these animal coronavirus infections in animals and humans.

[0082] The concept for treating coronavirus infections across all species is based on the structural similarities of coronaviruses. Therefore, it can be hypothesized that treatment and / or prevention options can be transferred from one coronavirus to another. Coronavirus particles contain four major structural proteins: spike (S), membrane (M), envelope (E), and nucleocapsid (N), all of which are encoded within the 3' end of the viral genome.

[0083] Coronaviruses contain a nonsegmented, positive-sense RNA genome of approximately 30 kb. The genome contains a 5' cap structure along with a 3' poly(A) tail, allowing it to act as mRNA for translation of the replicase polyprotein. The replicase genes encoding the nonstructural proteins (nsp) occupy two-thirds of the genome, approximately 20 kb, in contrast to the structural and accessory proteins, which comprise only approximately 10 kb of the viral genome. The coronavirus genome is organized as follows: 5'-leader-UTR-replicase-S (spike)-E (envelope)-M (membrane)-N (nucleocapsid)-3'UTR-poly(A) tail, with accessory genes interspersed within the structural genes at the 3' end of the genome. Although accessory proteins are not almost exclusively essential for replication in tissue culture, some have been shown to play important roles in viral pathogenesis (see Zhao et al. (2012) Cell Host Microbe 11:607-616).

[0084] The coronavirus life cycle begins with the initial attachment of the virion to a host cell through the interaction between the S protein and its receptor. The location of the receptor-binding domain (RBD) within the S1 region of the coronavirus S protein varies among viruses. The S protein-receptor interaction is a major determinant for coronavirus infection of host species and also governs the tissue tropism of the virus. Many coronaviruses utilize peptidases as their cellular receptors. The reason for the use of peptidases is unclear, as entry occurs even in the absence of the enzymatic domains of these proteins. Many alphacoronaviruses utilize aminopeptidase N (APN) as their receptor, while many betacoronaviruses, such as SARS-CoV-1, SARS-CoV-2, and HCoV-NL63, use the angiotensin-converting enzyme II (ACE2) receptor. MHV enters via CEACAM1, and MERS-CoV binds to dipeptidyl peptidase 4 (DPP4) to gain entry into human cells. After receptor binding, the virus must then gain access to the host cell cytosol. This is generally achieved by acid-dependent proteolytic cleavage of the S protein by a cathepsin, TMPRRS2 or another protease, followed by fusion of the viral membrane with the cellular membrane and ultimately release of the viral genome into the cytoplasm.

[0085] Coronaviruses encode either two or three proteases that cleave the replicase polyprotein: a papain-like protease (PL) encoded within nsp3, and a serine-type protease, the main protease, or M, encoded by nsp5. Most coronaviruses encode two PLs within nsp3, except for gammacoronaviruses, SARS-CoV-1, and MERS-CoV, which express only one PL (Mielech et al. (2014) Virus Res doi:10.1016).

[0086] This papain-like protease (PLpro) was found in SARS-CoV-1 to act similarly to the deubiquitinase in the human cellular ubiquitin proteasome system (UPS) (see Raaben et al. (2010) J Virol 84:7869-7879). The PLpro of SARS-CoV-2 shares a very high degree of homology with SARS-CoV-1 (96.1%, Nguyen et al. (2020) https: / / doi:org / 10:1101 / 2020.02.05.936013).

[0087] 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 all agents 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, and optionally at least one additional pharmaceutical drug listed below.

[0088] The term "excipient" is used herein to describe any component of a pharmaceutical composition other than the pharmaceutically active principle. The selection of a suitable excipient depends on various factors, such as the dosage form, dosage, desired solubility and stability of the composition.

[0089] 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.

[0090] 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.

[0091] The terms "treatment" and "therapy" include administering 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 ameliorate the disease course of coronavirus infection by completely curing the disease or by halting or slowing the progression of disability during the course of the disease.

[0092] The terms "prophylaxis" or "prophylactic treatment" 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, to prevent or inhibit the onset of symptoms resulting from coronavirus infection. 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.

[0093] The terms "subject" and "patient" include individuals suffering from a disease condition or disorder associated with a coronavirus infection, the diagnosis being either confirmed or suspected. The individual is a mammal, particularly a human.

[0094] Within the scope of this application, the term "medicine" is intended to include human and veterinary medicine.

[0095] 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its acceptable salts can be used as a monotherapy or can be further combined with at least one further active ingredient selected from the group comprising active ingredients used in disease-modifying therapy for coronavirus infections, symptomatic treatment of coronavirus infections and treatment of comorbidities.

[0096] For the effective treatment of coronavirus infection, it may be advantageous to provide combination therapy to patients in need thereof by combining 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts with at least one antiviral agent.

[0097] For example, from HIV, respectively, the following classes of antiretroviral therapy are known:

[0098] Suitable reverse transcriptase inhibitors for such combination therapy include nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). Examples of NRTIs include, but are not limited to, abacavir, didanosine, emtricitabine, lamivudine, stavudine, tenofovir, zidovudine, zalcitabine, entecavir, adefovir, elvucitabine, fosalvudine (-tidoxil), fozivudine tidoxil, radiciclovir, alamifovir, clevudine, pradefovir, and telbivudine. Examples of NNRTIs include, but are not limited to, efavirenz, etravirine, nevirapine, rilpivirine, delavirdine, emivirine, and lersivirine.

[0099] Suitable for combination therapy according to the present invention are integrase inhibitors such as raltegravir, elvitegravir, dolutegravir, MK-2048.

[0100] Examples of HIV protease inhibitors suitable for combination therapy according to the present invention are saquinavir, indinavir, ritonavir, nelfinavir, amprenavir, lopinavir, atazanavir, fosamprenavir, tipranavir, darunavir, brecanavir, mozenavir, tipranavir.

[0101] Examples of entry inhibitors suitable for combination therapy according to the present invention are enfuvirtide and maraviroc.

[0102] Further common viral inhibitors suitable for combination therapy according to the present invention include ancriviroc, aplaviroc, cenicriviroc, enfuvirtide, maraviroc, vicriviroc, amantadine, rimantadine, pleconaril, idoxuridine, acyclovir, brivudine, famciclovir, penciclovir, sorivudine, valacyclovir, cidofovir, ganciclovir, valganciclovir, sofosbuvir, foscarnet, ribavirin, taribavirin, and filbuvir. , nesbuvir, tegovir, fosdevirine, favipiravir, avifavir, merimeposib, asunaprevir, valapiravir, bocepovir, cilprevir, danoprevir, daclatasvir, narulaprevir, telaprevir, simeprevir, vanipevir, rupintrivir, remdesivir, fomivirsen, amenamevir, alisporivir, bevirimat, letermovir, laninamavir, oseltamivir, peramivir, zanamivir.

[0103] Common immunostimulants suitable for combination therapy according to the invention can be selected from the group comprising interferons (alpha-, beta-, gamma-, tau-interferons), interleukins, CSF, PDGF, EGF, IGF, THF, levamisole, dimepranol, inosine.

[0104] Additionally, possible combinations according to the present invention include adjuvants such as cobicistat.

[0105] The comorbidities may be due to or independent of the disorders caused by coronavirus infection. Thus, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts can be combined with at least one additional active ingredient selected from the group consisting of steroidal and nonsteroidal anti-inflammatory drugs, immunomodulators, immunosuppressants, anti-infectives such as antibiotics, antiretrovirals, antivirals, antifungals, and antiprotozoal agents, analgesics, anticoagulants, antiplatelet agents, bronchodilators, pulmonary vasodilators, mucolytic agents, pulmonary surfactants, antioxidants, ENaC activators, HMG-CoA reductase inhibitors, or AT1 receptor antagonists for use in the prevention or treatment of coronavirus infection.

[0106] Suitable examples of such steroidal anti-inflammatory drugs include corticosteroids, glucocorticoids, cortisone, cortisone acetate, hydrocortisone, hydrocortisone acetate, dexamethasone, betamethasone, prednisone, prednisolone, methylprednisolone, deltasone, triamcinolone, tixocortol pivalate, mometasone, amcinonide, budesonide, desonide, fluoconide, fluocinolone, halcinonide, flucortolone, hydrocortisone-17-valerate, halometasone, and alkoxydipropionate. Includes lometasone, betamethasone valerate, betamethasone dipropionate, prenicarbate, clobetasone-17-butyrate, clobetasol-17-propionate, flucortolone caproate, flucortolone pivalate, fluiden acetate, hydrocortisone-17-butyrate, hydrocortisone-17-aceponate, hydrocortisone-17-buteprate, ciclesonide, flunisolide, fluticasone furoate, fluticasone propionate, triamcinolone acetonide, and beclomethasone dipropionate.

[0107] Suitable examples of such nonsteroidal anti-inflammatory drugs (NSAIDs) include acetylsalicylic acid, salicylic acid and salicylates, acetaminophen (paracetamol), salsalate, diflunisal, ibuprofen, dexibuprofen, naproxen, fenoprofen, ketoprofen, dexketoprofen, flurbiprofen, oxaprozin, loxoprofen, indomethacin, tolmetin, sulindac, etodolac, ketorolac, diclofenac, benzodiazepine, benzocaine ... Includes phenac, aceclofenac, nabumetone, piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, isoxicam, phenylbutazone, mefenamic acid, meclofenamic acid, flufenamic acid, tolfenamic acid, celexoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, flocoxib, nimesulide, clonixin, licofelone, H-harpagide, flunixin, and tiaprofenic acid.

[0108] Suitable examples of such immunomodulatory agents include, among others, thalidomide, lenalidomide, pomalidomide and apremilast.

[0109] Suitable examples of such immunosuppressants include glucocorticoids as listed above; cytostatics such as alkylating agents (such as cyclophosphamide), antimetabolites such as methotrexate, azathioprine, mercaptopurine, fluorouracil, leflunomide, and the like; protein synthesis inhibitors and certain antibiotics such as dactinomycin, anthracyclines, mitomycin C, bleomycin, and mithramycin; intercalating agents such as mitoxantrone; muromonab-CD3, rituximab, ustekinumab, and the like. antibodies such as riboflavin, alemtuzumab, natalizumab, basiliximab, tocilizumab and daclizumab; drugs acting on immunophilins such as cyclosporine, tacrolimus and sirolimus, non-classified immunosuppressants such as beta-interferon and gamma-interferon, opioids, TNF-binding proteins such as infliximab, etanercept, adalimumab; or the group of curcumin, catechin, mycophenolic acid, fingolimod, myriocin and fumaric acid dimethyl ester.

[0110] Anti-infective agents are a general term for compounds useful in the treatment of bacterial, viral, fungal, and parasitic infections (e.g., protozoal or parasitic), and include antibiotics, antifungals, antiprotozoal agents, and the aforementioned antivirals.

[0111] Suitable examples of such antibiotics include imipenem, meropenem, ertapenem, cephalosporins, aztreonam, penicillins such as penicillin G and penicillin V, piperacillin, mezlocillin, ampicillin, amoxicillin, flucloxacillin, methicillin, oxacillin, clavulanic acid, sulbactam, tazobactam, sultamicillin, fosfomycin, teicoplanin, vancomycin, bacitracin, colistin, gramicidin, polymyxin B, tyrothricin, teixobactin, fosmidomycin, amikacin, gentamicin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin, chloramphenicol, fusiforme ... These include: benzodiazepine, cethromycin, narbomycin, telithromycin, clindamycin, lincomycin, daptomycin, dalfopristin, quinupristin, azithromycin, clarithromycin, erythromycin, roxithromycin, linezolid, doxycycline, minocycline, tetracycline, oxytetracycline, tigecycline, norfloxacin, enoxacin, ciprofloxacin, ofloxacin, levofloxacin, moxifloxacin, metronidazole, tinidazole, aminocoumarin, sulfadiazine, sulfadoxine, sulfamethoxazole, sulfasalazine, pyrimethamine, trimethoprim, and rifampin.

[0112] Suitable examples of such antifungal (antifungal) drugs are abafungin, amphotericin B, candicidin, filipin, hamycin, natamycin, nystatin, rimocidin, bifonazole, butoconazole, clotrimazole, econazole, fenticonazole, isoconazole, ketoconazole, luliconazole, miconazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, albaconazole, efinaconazole, efinaconazole, erico ... These include poxiconazole, fluconazole, isavuconazole, itraconazole, posaconazole, propiconazole, ravuconazole, terconazole, voriconazole, amorolifine, butenafine, nafitifine, terbinafine, anidulafungin, caspofungin, micafungin, benzoic acid, ciclopirox, flucytosine, griseofulvin, haloprogin, tolnaftate, undecylenic acid, crystal violet, and balsam of Peru.

[0113] Suitable examples of such antiprotozoal agents include metronidazole, tinidazole, ornidazole, atovaquone, clioquinol, chlorquinaldol, emetine, pentamidine isethionate, eflornithine, nitrofural, halofuginone, miltefosine, chloroquine, hydroxychloroquine, mepacrine, primaquine, amodiaquine, pamaquine, piperaquine, proguanil, cyclohexanone, quinine, mefloquine, pyrimethamine, artemether, artemisinin, artesunate, dihydroartemisinin, halofantrine, lemantrine, and sulfadoxine.

[0114] Suitable examples of further antiparasitic agents include meglumine antimoniate, benznidazole, sodium stibogluconate, fumagillin, halofantrine, melarsoprol, nifurtimox, nitazoxanide, permethrin, lindane, malathion, carbaryl, pyrethrim, fenothrin, bioallethrin, imidacloprid, moxidectin, nitenpyram, fipronil, pyriprol, selamectin, dimpyrate, spinosad, indoxacarb, methoprene, pyriproxyfen, lufenuron, neem oil, citronella oil, clove oil, peppermint oil, eucalyptus oil.

[0115] Suitable examples of analgesics include the NSAIDs listed above; opioid analgesics such as morphine, fentanyl, methadone, oxycodone, carfetanil, dihydroetorphine, omefentanil, etorphine, sufentanil, remifentanil, alfentanil, buprenorphine, hydromorphone, levomethadone, hydrocodone, pintramide, nalbuphine, tapentadol, pentazocine, dihydrocodeine, codeine, pethidine, tramadol, tilidine, meptazinol, naloxone, naltrexone, diprenorphine, loperamide, apomorphine; epibatidine; scopolamine; ziconit; cannabinoids such as tetrahydrocannabinol, cannabidiol, and marinol; flupirtine; ketamine, and the local anesthetics listed above.

[0116] Suitable examples of such anticoagulants include heparin, coumarins such as phenprocoumon (marcumar) and warfarin, apixaban, rivaroxaban, edoxaban, dabigatran, ximelagatran, hirudin, lepirudin, bivalirudin, citrate, EDTA, fondaparinux, argatroban, otamixaban.

[0117] Suitable examples of such antiplatelet agents include abciximab, acetylsalicylic acid, dipyridamole, clopidogrel, eptifibatide, ilomedin, prostacyclin, prasugrel, ticagrelor, ticlopidine and tirofiban.

[0118] Suitable bronchodilators, such as beta-2 adrenergic receptor agonists, include short-acting beta-2 agonists (SABAs), such as salbutamol, albuterol, bitolterol, fenoterol, isoprenaline, levosalbutamol, levalbuterol, orciprenaline, pirbuterol, procaterol, ritodrine, and terbutaline; long-acting beta-2 agonists (LABAs), such as arformoterol, bambuterol, clenbuterol, formoterol, and salmeterol; ultra-long-acting beta-2 agonists, such as besiterol, carmoterol, indacaterol, olodaterol, and vilanterol, alone or in combination with umeclidinium bromide and / or fluticasone furoate; beta-2 agonists of unknown duration of action such as isoxsuprine, mabuterol, or zilpaterol.

[0119] Suitable muscarinic anticholinergics (bronchodilatory M3 receptor antagonists) include ipratropium bromide, tiotropium bromide, oxitropium bromide, glycopyrronium bromide, aclidinium bromide, umeclidinium bromide, atropine, hyoscyamine, aclidinium bromide, 4-DAMP, darifenacin, DAU-5884, HL-031, HL-120, J-104, J-129, procyclidine, oxybutynin, tolterodine and zamifenacin.

[0120] Further bronchodilators include epinephrine, ephedrine, theophylline and TSG12.

[0121] A potent pulmonary vasodilator is nitric oxide. Further suitable pulmonary vasodilators are prostacyclin (prostaglandin PGI2) analogues, such as iloprost, epoprostenol and treprostinil.

[0122] Suitable mucolytic agents include N-acetylcysteine ​​(NAC), ambroxol, bromhexine, carbocysteine, erdocysteine, mecysteine, and dornase alfa.

[0123] Suitable pulmonary surfactants include synthetic compositions such as colfosceril palmitate, Pumactant, KL-4, Venticute, and Lucinactant, as well as animal-derived surfactants such as Beractant, Calfactant, and Poractant alfa.

[0124] A powerful antioxidant is inhaled carbon monoxide (CO).

[0125] Suitable ENaC (epithelial sodium channel) activating peptides include AP301 and S3969.

[0126] Suitable HMG-CoA reductase inhibitors (statins) include atorvastatin, alone or in combination with amlodipine and / or perindopril, cerivastatin, fluvastatin, lovastatin, alone or in combination with niacin, mevastatin, pitavastatin, pravastatin, rosuvastatin, alone or in combination with ezetimibe, simvastatin, alone or in combination with ezetimibe or niacin.

[0127] Suitable AT1 antagonists (angiotensin II receptor blockers; tartrates) include losartan, valsartan, candesartan, telmisartan, irbesartan, olmesartan, eprosartan, fimasartan, azilsartan, mirfasartan, pomisartan, pratosartan, lipisartan, tasosartan, saprosartan and EXP3174.

[0128] One of 5-amino-2,3-dihydro-1,4-phthalazinedione or its pharmaceutically acceptable salt and the additional active ingredient can be used simultaneously, separately, or sequentially to treat or prevent disease symptoms. The two active ingredients can be provided in a single dosage form or as separate formulations, each formulation containing at least one of the two active ingredients. One or both of the two active ingredients can be formulated as a bolus.

[0129] In particular, the present application discloses 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the treatment of coronavirus infections, which have been refractory to previous treatment with at least one other pharmaceutically active agent.

[0130] The term "medicine" or "healthcare" includes human and veterinary medicine.

[0131] The term "organism" refers to an organism, particularly a human or animal, that has an autoregulatory immune system.

[0132] The term "host organism" is used in reference to the organism utilized by a virus, and herein particularly a retrovirus, for replication after infection by the virus.

[0133] The term "active agent" in this application, unless otherwise specified, refers to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts. In addition, this term may include further pharmaceutical agents known from the state of the art.

[0134] The terms "composition" and "pharmaceutical composition" include 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts in any pharmacologically suitable defined dose and dosage form together with at least one suitable excipient or carrier substance, and all substances that are produced directly or indirectly as a combination, accumulation, complex formation or crystallization of the above-mentioned components or result from other reactions or interactions, and optionally at least one further pharmaceutical agent known in the state of the art.

[0135] The term "excipient" is used in this application to describe each component of a pharmaceutical composition in addition to the active agent. The selection of a suitable excipient depends on factors such as the dosage form and dose, as well as the excipient's own effect on the solubility and stability of the composition.

[0136] The term "operation" describes the unique and specific mode of operation of each agent within the scope of this application.

[0137] The terms "effect," "therapeutic effect," "action," and "therapeutic effect" with respect to at least one active agent according to the present invention refer to a beneficial result that causally occurs to an organism to which the at least one active agent is administered.

[0138] In the context of this application, "therapeutically effective dose" means that a sufficient dose of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts is administered to an organism or patient in need of such treatment.

[0139] The terms "joint administration," "combined administration," or "simultaneous administration" of at least one pharmaceutical agent according to the present invention and / or at least one pharmaceutical agent from the state of the art include administration of the mentioned agents at the same time or at times substantially related to each other, as well as administration of the agents at different times within a coherent experiment. The chronological order of administration of the agents is not limited by these terms. A person skilled in the art will have no difficulty in estimating the described administration in terms of their chronological order or local order from his / her knowledge and experience.

[0140] The term "organism" refers to any animal, particularly a vertebrate, including a human. A "patient" in the context of this application is an organism that is afflicted with a definable and diagnosable disease and to which a suitable active agent can be administered.

[0141] The terms "prophylaxis," "treatment," and "therapy" include the administration of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts to an organism, either alone or in combination with at least one additional pharmaceutical agent known in the art, to prevent the onset of a particular disease, inhibit or alleviate the symptoms, or initiate the healing process of the respective disease.

[0142] 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, the composition according to the present invention or the drug combination according to the present invention can be applied for the prevention or treatment of coronavirus infections by any medically acceptable route of administration to patients in need thereof. Such medically acceptable routes of administration may be, for example, by inhalation, by intubation, orally, parenterally, intraperitoneally, intravenously, intraarterially, intramuscularly, topically, transdermally, subcutaneously, intradermally, sublingually, conjunctivally, intravaginally, rectally or nasally.

[0143] A preferred oral formulation for use in the prevention or treatment of coronavirus infection is a capsule or tablet containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts in an amount of 50 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg, 500 mg or 600 mg, preferably 100 mg, 150 mg, 200 mg, 300 mg or 400 mg, most preferably 300 mg.

[0144] In another aspect of the present invention, a composition for use in the prevention or treatment of a coronavirus infection is disclosed, the composition comprising 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a carrier, and at least one pharmaceutically acceptable excipient.

[0145] The term "pharmaceutically acceptable excipient" refers to natural or synthetic compounds added to pharmaceutical formulations along with pharmaceutically active agents. They may help to bulk the formulation, improve the formulation's desired pharmacokinetic properties or stability, and be beneficial in the manufacturing process. Advantageous classes of excipients according to the present invention include carriers, binders, colorants, buffers, preservatives, antioxidants, coatings, sweeteners, thickeners, pH adjusters, acidity adjusters, acidifiers, solvents, tonicity adjusters, disintegrants, glidants, lubricants, emulsifiers, solubilizers, stabilizers, diluents, anti-caking agents (anti-adherents), adsorbents, foaming agents, anti-foaming agents, opacifiers, fats, viscosity enhancers, hydrotropes, fragrances, and flavoring substances.

[0146] Generally, one or more pharmaceutically acceptable carriers are added to pharmaceutically active agents.Eligible are all carriers known in the art and their combinations.For solid dosage forms, they can be, for example, plant and animal fats, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talcum, zinc oxide.For liquid dosage forms and emulsions, suitable carriers are, for example, solvents, solubilizers, emulsifiers, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol, cottonseed oil, peanut oil, olive oil, castor oil, sesame oil, glycerol fatty acid esters, polyethylene glycol, fatty acid esters of sorbitan. Suspensions according to the present invention may use carriers known in the art, such as diluents (e.g., water, ethanol or propylene glycol), ethoxylated isostearyl alcohol, polyoxyethylene and polyoxyethylene sorbitan esters, microcrystalline cellulose, bentonite, agar, tragacanth, and the like.

[0147] The term binder refers to a substance that binds or glues powders together, making them cohesive through granulation. They function as the "adhesive" of the formulation. Binders increase the cohesive strength of the diluent or filler provided.

[0148] Suitable binders are, for example, starch from wheat, corn, rice or potato, gelatin, naturally occurring sugars such as glucose, sucrose or beta-lactose, sweeteners from corn, natural and synthetic gums such as acacia, tragacanth or calcium ammonium alginate, sodium alginate, carboxymethylcellulose, sodium carboxymethylcellulose, hydroxypropyl carboxymethylcellulose, polyethylene glycol, polyvinylpyrrolidone, magnesium aluminum silicate, waxes, etc. The percentage of binder in the composition may range from 1 to 30% by weight, preferably from 2 to 20% by weight, more preferably from 3 to 10% by weight, most preferably from 3 to 6% by weight.

[0149] 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. An additional benefit of colorants is that they can make spilled aqueous solutions on the nebulizer 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.

[0150] Suitable pharmaceutical colorants are, for example, curcumin, riboflavin, riboflavin-5'-phosphate, tartrazine, alkannin, 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, caramel, caustic sulfite caramel, ammonia caramel. Benzyl alcohol, ammonium sulfite 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.

[0151] Furthermore, buffer solutions are preferred for liquid formulations, especially pharmaceutical liquid formulations. The terms buffer, buffer system, and buffer solution, especially for aqueous solutions, refer to the ability of the system to resist pH changes due to the addition of an acid or base or 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), diethylmalonate, 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:

[0152] Carbonate buffers such as acetate buffers and dicarboxylic acid buffers such as fumarate, tartrate and phthalate, and tricarboxylic acid buffers such as citrate are preferred.

[0153] 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. More 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), MOPS, and N,N-bis-(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES). Another group of preferred buffers is glycine, glycyl-glycine, glycyl-glycyl-glycine, N,N-bis-(2-hydroxyethyl)glycine, and N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine (tricine). 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 their derivatives are also preferred. KH2PO4 buffer is particularly preferred.

[0154] Preservatives for liquid and / or solid dosage forms may be used as required, including sorbic acid, potassium sorbate, sodium sorbate, calcium sorbate, methylparaben, ethylparaben, methylethylparaben, propylparaben, benzoic acid, sodium benzoate, potassium benzoate, calcium benzoate, heptyl p-hydroxybenzoate, methyl parahydroxybenzoate, sodium ethyl parahydroxybenzoate, sodium propyl parahydroxybenzoate, benzyl alcohol, benzalkonium chloride, phenylethyl alcohol, cresol, cetylpyridinium chloride, chlorbutanol, thiomersal (sodium 2-(ethylmercurithio)benzoate), sulfur dioxide, sodium sulfite, sodium bisulfite, sodium metabisulfite, potassium metabisulfite, potassium sulfite, calcium sulfite, calcium bisulfite. The antioxidant may be selected from the group including, but not limited to, methylparaben, potassium bisulfite, biphenyl, orthophenylphenol, orthophenylphenol sodium, thiabendazole, herring, 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.

[0155] The addition of a sufficient amount of antioxidant is particularly preferred for liquid and topical formulations.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.The use of sodium metabisulfite, alpha-tocopherol and ascorbyl palmitate is preferred.

[0156] Tablets or pills are usually coated, i.e., the coating constitutes the outer layer. This can be a film coating, a sugar coating containing sugars, or a compression coating. Pharmaceutically acceptable varnishes or waxes, such as HPMC (hydroxypropyl methylcellulose), MC (methylcellulose), or HPC (hydroxypropyl cellulose), can be used. Such coatings can help disguise the taste and facilitate swallowing or identification. Plasticizers and pigments are often included in the coating. Capsules usually have a gelatinous envelope that encapsulates the active substance. The specific composition and thickness of this gelatinous layer determine how quickly absorption occurs after ingestion of the capsule. As is known in the art, sustained-release formulations are of particular interest.

[0157] Suitable sweeteners may be selected from the group comprising mannitol, glycerol, acesulfame potassium, aspartame, cyclamate, isomalt, isomaltitol, saccharin and its sodium, potassium and calcium salts, sucralose, alitame, thaumatin, glycyrrhizin, neohesperidin dihydrochalcone, steviol glycosides, neotame, aspartame-acesulfame salt, maltitol, maltitol syrup, lactitol, xylitol, erythritol.

[0158] Suitable thickening agents may be selected from the group comprising, but not limited to, polyvinylpyrrolidone, methylcellulose, hydroxypropyl methylcellulose, hydroxypropyl cellulose, dextrin, polydextrose, modified starch, alkali modified starch, bleached starch, oxidized starch, enzyme treated starch, monostarch phosphate, distarch phosphate esterified with sodium trimetaphosphate or phosphorus oxychloride, distarch phosphate, acetylated distarch phosphate, starch acetate esterified with acetic anhydride, starch acetate esterified with vinyl acetate, acetylated distarch adipate, acetylated distarch glycerol, distarch glycerin, hydroxypropyl starch, hydroxypropyl distarch glycerin, hydroxypropyl distarch phosphate, hydroxypropyl distarch glycerol, starch sodium octenylsuccinate, acetylated oxidized starch, hydroxyethyl cellulose.

[0159] 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.

[0160] Suitable acidity regulators include acetic acid, potassium acetate, sodium acetate, sodium diacetate, calcium acetate, carbon dioxide, malic acid, fumaric acid, sodium lactate, potassium lactate, calcium lactate, ammonium lactate, magnesium lactate, citric acid, mono-, di-, trisodium citrate, mono-, di-, tripotassium citrate, mono-, di-, tricalcium citrate, tartaric acid, mono-, disodium tartrate, mono-, dipotassium tartrate, potassium sodium tartrate, orthophosphoric acid, lecithin citrate, magnesium citrate, ammonium malate, sodium malate, sodium hydrogen malate, calcium malate, calcium hydrogen malate, adipic acid, adipic acid, The surfactant may be selected from the group comprising sodium phosphate, potassium adipate, ammonium adipate, succinic acid, sodium fumarate, potassium fumarate, calcium fumarate, ammonium fumarate, 1,4-heptonolactone, triammonium citrate, diammonium citrate, calcium glycerophosphate, isopropyl citrate, potassium carbonate, potassium bicarbonate, ammonium carbonate, ammonium bicarbonate, magnesium carbonate, magnesium bicarbonate, ferrous carbonate, ammonium sulfate, aluminum potassium sulfate, aluminum ammonium sulfate, sodium hydroxide, potassium hydroxide, ammonium hydroxide, magnesium hydroxide, gluconic acid.

[0161] Acidifying agents are used which are inorganic chemicals that produce or become acids. Suitable examples are ammonium chloride and calcium chloride.

[0162] 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.

[0163] Suitable tonicity 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.

[0164] Suitable disintegrants can be selected from the group consisting of starch, cold-soluble starches such as carboxymethyl starch, cellulose derivatives such as methylcellulose and sodium carboxymethylcellulose, cross-linked microcrystalline cellulose such as microcrystalline cellulose and croscarmellose sodium, natural and synthetic gums such as guar, agar, Karaya (Indian tragacanth), locust bean gum, and tragacanth, clays such as bentonite, xanthan gum, alginates such as alginic acid and sodium alginate, and effervescent compositions. Hydration is supported by, for example, starch, cellulose derivatives, alginates, polysaccharides, dextran, and cross-linked polyvinylpyrrolidone. The amount of disintegrant in the composition can vary from 1 to 40% by weight, preferably from 3 to 20% by weight, and most preferably from 5 to 10% by weight.

[0165] Lubricants are materials that prevent the individual supplements from sticking and improve the flow characteristics of the granulation so that the flow is smooth and consistent. Suitable glidants include silicon dioxide, magnesium stearate, sodium stearate, starch, and talc. The amount of glidant in the composition can vary from 0.01 to 10% by weight, preferably from 0.1 to 7% by weight, more preferably from 0.2 to 5% by weight, and most preferably from 0.5 to 2% by weight.

[0166] The term "lubricant" refers to a substance added to a dosage form to facilitate the release of tablets, granules, etc. from a press die or exit nozzle. They reduce friction or wear. Lubricants are usually added immediately before pressing, as they should be present on the surface of the granules and between the granules and the press die parts. The amount of lubricant in a composition can vary from 0.05 to 15% by weight, preferably 0.2 to 5% by weight, more preferably 0.3 to 3% by weight, and most preferably 0.3 to 1.5% by weight. Suitable lubricants are, among others, metal stearates such as sodium oleate, sodium stearate, calcium stearate, potassium stearate, and magnesium stearate, stearic acid, sodium benzoate, sodium acetate, sodium chloride, boric acid, high-melting waxes, and polyethylene glycol.

[0167] The emulsifier can be chosen, for example, from the following anionic and nonionic emulsifiers: anionic emulsifier wax, cetyl alcohol, cetylstearyl alcohol, stearic acid, oleic acid, polyoxyethylene polyoxypropylene block polymers, addition products of 2 to 60 mol of ethylene oxide onto castor oil and / or hydrogenated castor oil, wool wax oil (lanolin), sorbitan esters, polyoxyethylene alkyl esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethene sorbitan monolaurate. , Polyoxyethene sorbitan monooleate, Polyoxyethene sorbitan monopalmitate, Polyoxyethene sorbitan monostearate, Polyoxyethene sorbitan tristearate, Polyoxyethene stearate, Polyvinyl alcohol, Metatartaric acid, Calcium tartrate, Alginic acid, Sodium alginate, Potassium alginate, Ammonium alginate, Calcium alginate, Propane-1,2-diol alginate, Carrageenan, Modified Eucheuma seaweed, Locust bean gum, Tragacanth, Acacia gum, Karaya gum, Ge Langum, Ghatti Gum, Glucomannan, Pectin, Amidated Pectin, Ammonium Phosphatide, Brominated Vegetable Oil, Sucrose Acetate Isobutyrate, Glycerol Ester of Wood Rosin, Disodium Phosphate, Trisodium Diphosphate, Dicalcium Diphosphate, Dicalcium Dihydrogen Phosphate, Sodium Triphosphate, Pentapotassium Triphosphate, Sodium Polyphosphate, Calcium Polyphosphate, Ammonium Polyphosphate, Beta-Cyclodextrin, Powdered Cellulose, Methylcellulose, Ethylcellulose, Hydroxypropylcellulose, Hydroxypropylmethylcellulose, Ethylmethylcellulose, Carboxymethylcellulose, Sodium Carboxymethylcellulose, Ethylhydroxyethylcellulose, Croscarmellose, Enzymatically Hydrolyzed Carboxymethylcellulose, Mono- and Diglycerides of Fatty Acids, Glyceryl Monostearate, Glyceryl Distearate, Acetate Esters of Mono- and Diglycerides of Fatty Acids, Lactic Acid Esters of Mono- and Diglycerides of Fatty Acids, Citric Acid Esters of Mono- and Diglycerides of Fatty Acids, Tartaric Acid Esters of Mono- and Diglycerides of Fatty Acids,Mono- and diacetyl tartaric acid esters of mono- and diglycerides of fatty acids, mono- and diacetyl tartaric acid esters of mono- and diglycerides of fatty acids, mixed acetic and tartaric acid esters of mono- and diglycerides of fatty acids, succinylated monoglycerides, sucrose esters of fatty acids, sucroglycerides, polyglycerol esters of fatty acids, polyglycerol polyricinoleate, propane-1,2-diol esters of fatty acids, propylene glycol esters of fatty acids, lactylated fatty acid esters of glycerol and propane-1, thermally oxidized soybean oil interacting with mono- and diglycerides of fatty acids, sodium dioctyl sulfosuccinate, sodium stearoyl-2-lactylate, calcium stearoyl-2-lactylate, alcohol Stearyl tartarate, stearyl citrate, sodium stearyl fumarate, calcium stearoyl fumarate, stearyl tartrate, stearyl citrate, sodium stearyl fumarate, calcium stearoyl fumarate, sodium lauryl sulfate, ethoxylated mono- and diglycerides, methyl glucoside-coconut oil ester, sorbitan monostearate, sorbitan tristearate, sorbitan monolaurate, sorbitan monooleate, sorbitan monopalmitate, sorbitan trioleate, sodium calcium polyphosphate, calcium polyphosphate, ammonium polyphosphate, cholic acid, choline salts, didarch glycerol, starch sodium octenylsuccinate, acetylated oxidized starch. Glycerin monooleate, stearic acid, and phospholipids such as lecithin are preferred.

[0168] Suitable surface-active solubilizers (solubilizers) are, for example, diethylene glycol monoethyl ester, polyethylpropylene glycol copolymers, cyclodextrins, such as α- and β-cyclodextrin, glyceryl monostearate, for example Solutol HS 15 (Macrogol-15-hydroxystearate, PEG 660-15 hydroxystearate from BASF), sorbitan esters, polyoxyethylene glycol, polyoxyethylene sorbitan acid esters, polyoxyethylene sorbitan monooleate, polyoxyethylene oxystearic acid triglyceride, polyvinyl alcohol, sodium dodecyl sulfate, (anionic) glyceryl monooleate.

[0169] Stabilizers are substances that can be added to prevent undesired changes. Although stabilizers are not actual emulsifiers, they can also contribute to the stability of emulsions. Suitable examples of stabilizers include oxystearin, xanthan gum, agar, oat gum, guar gum, tara gum, polyoxyethene stearate, aspartame-acesulfame salt, amylase, protease, papain, bromelain, ficin, invertase, polydextrose, polyvinylpyrrolidone, polyvinylpolypyrrolidone, triethyl citrate, maltitol, and maltitol syrup.

[0170] Diluents or fillers are inert substances added to a drug to handle a minimal amount of active agent. Examples of suitable diluents are water, mannitol, pregelatinized starch, starch, microcrystalline cellulose, powdered cellulose, silicified microcrystalline cellulose, dibasic calcium phosphate dihydrate, calcium phosphate, calcium carbonate, hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, polyethylene glycol, xanthan gum, gum arabic, or any combination thereof.

[0171] Anti-caking agents (anti-adherents) can be added to the supplement or supplement composition to prevent the formation of lumps and facilitate packaging, transportation, release from at least one chamber of the dispensing cap, and consumption. Suitable examples include tricalcium phosphate, powdered cellulose, magnesium stearate, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, bone phosphate, sodium silicate, silicon dioxide, calcium silicate, magnesium trisilicate, talc powder, sodium aluminosilicate, potassium aluminum silicate, calcium aluminosilicate, bentonite, aluminum silicate, stearic acid, and polydimethylsiloxane.

[0172] Sorbents are materials that absorb oil from water. Suitable examples include natural sorbents, such as peat moss, sawdust, feathers, and any natural sorbents containing carbon, as well as synthetic sorbents such as polyethylene and nylon. Sorbents are used to protect tablets / capsules from moisture by limited fluid adsorption (the uptake of liquids or gases by adsorption or sorption) in a dry state.

[0173] In some galenical preparations, it may be desirable for liquid oral dosage forms to produce some foam upon dissolution. Such an effect can be supported by the addition of foaming agents, which reduce the surface tension of the liquid and thus promote the formation of bubbles, or increase its colloidal stability by inhibiting bubble coalescence. Alternatively, foam may be stabilized. Suitable examples include mineral oil, Quillaja extract, triethyl citrate, sodium lauryl ether sulfate, sodium lauryl sulfate, and ammonium lauryl sulfate.

[0174] Alternatively, some liquid oral dosage forms may appear slightly foamy when prepared. This does not interfere with the desired use, but it may affect patient compliance in the case of pharmaceuticals or commercial success in the case of dietary supplements. Therefore, it may be desirable to add a pharmaceutically acceptable antifoaming agent (defoamer). Examples are polydimethylsiloxane or silicone oil in dietary supplements, or simethicone in pharmaceuticals.

[0175] 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.

[0176] Suitable fats are, for example, decyl oleate, hydrated castor oil, light mineral oil, mineral oil, polyethylene glycol, sodium lauryl sulfate.

[0177] Consistency improvers are, for example, cetyl alcohol, cetyl ester wax, hydrated castor oil, microcrystalline wax, nonionic emulsifier wax, beeswax, paraffin or stearyl alcohol.

[0178] Suitable hydrotropes are alcohols such as ethanol, isopropyl alcohol, or polyols such as glycerin.

[0179] Suitable aromatic and flavoring substances include, inter alia, essential oils, which can be used for this purpose. Generally, this term refers to volatile extracts from plants or plant parts, each of which has a characteristic odor. They can be extracted from plants or plant parts by steam distillation.

[0180] Suitable examples are essential oils, respectively rapeseed, sage, cedar, clove, chamomile, anise, aniseed, star anise, thyme, tea tree, peppermint, mint oil, menthol, cineole, borneol, gingerol, eucalyptus, mango, fig, lavender oil, chamomile flower, pine needle, cypress, orange, rose, rosewood, plum, quince, cherry, birch leaf, cinnamon, lime, grapefruit, tangerine, zucchini, jasmine, jasmine, jasmine, jasmine oil ... Juniper, valerian, lemon, lemon rose, lemongrass, palmarosa, cranberry, pomegranate, rosemary, ginger, pineapple, guava, echinacea, ivy extract, blueberry, persimmon, melon, alpha- or beta-pinene, alpha-pinene oxide, alpha-campholenaldehyde, alpha-citronellol, alpha-isoamyl-cinnamic acid, alpha-terpinene cinnamate, alpha-terpineol, alpha-terpinene, aldehyde C 16, alpha-phellandrene, amyl cinnamic aldehyde, amyl salicylate, anisaldehyde, basil, anethole, bay, benzyl acetate, benzyl alcohol, bergamot, bitter orange peel, black pepper, calamus, camphor, cananga oil, cardamom, carnation, carvacrol, carveol, cassia, castor, cedarwood, cinnamic aldehyde, cinnamic alcohol, cis-pinane, citral, citronella, citronellal, citronellol dextro, citronellol, citronellyl acetate; citronellyl nitrile, citrus unshiu, clary sage, clove bud, coriander, corn, cottonseed, d-dihydrocarvone, decyl aldehyde, diethyl phthalate, dihydroanethole, dihydrocarveol, dihydrolinalool, dihydromyrcene, dihydromyrcenol, dihydromyrcenyl acetate Dihydroterpineol, dimethyl salicylate, dimethyloctanal, dimethyloctanol, dimethyloctanyl acetate, diphenyl oxide, dipropylene glycol, d-limonene, d-pulegone, estragole, ethyl vanillin, eucalyptol, Eucalyptus citriodora, Eucalyptus groves, eugenol, evening primrose, fencol, fennel, ferniol, fish, florasone, galaxolide, geraniol, geranium, geranyl acetate, geranyl nitrile, guaiacol, guaiac wood, Gurjun balsam, heliotropin, erbanate, hiba, hydroxynitroneral, i-carvone, i-methyl acetate, ionone, isobutylquinolein, isobornyl acetate, isobornyl methyl ether, isoeugenol, isolongifolene, jasmine, lavender, limonene, linalool oxideoxide), linallol, linalyl acetate, flaxseed, Lithospermum camphorata, I-methyl acetate, longifolene, mandarin, mint, menthane hydroperoxide, menthol crystals, menthol levo, menthone levo, methyl anthranilate, methyl cedryl ketone, methyl chavicol, methyl hexyl ether, methyl ionone, methyl salicylate, minerals, mint, musk ambrette, musk ketone, musk xylose , myrcene, nerol, neryl acetate, nonyl aldehyde, nutmeg, orris root, para-cymene, para-hydroxyphenylbutanone crystals, patchouli, p-cymene, pentyl oil, pepper, perillaldehyde, petitgrain, phenylethyl alcohol, phenylethyl propionate, phenylethyl-2-methylbutyrate, pimento berry, pimento leaf, pinan hydroperoxide, pinanol, pine esters, pine, pinene, piperona aromatic substances from menthol, piperonyl acetate, piperonyl alcohol, purinol, purinyl acetate, pseudoionone, rhodinol, rhodinyl acetate, rosalin, rue, sandalwood, sandenol, sassafras, sesame, soybean, spearmint, spices, spike lavender, spilanthol, starflower, tea seed, terpenoids, terpineol, terpinolene, terpinyl acetate, tert-butylcyclohexyl acetate, tetrahydrolinalool, tetrahydrolinalyl acetate, tetrahydromyrcenol, tulasi, thymol, tomato, trans-2-hexenol, trans-anethole, turmeric, turpentine, vanillin, vetiver, vitalizer, white cedar, white grapefruit, wintergreen, etc., or mixtures thereof, and mixtures of menthol, peppermint, and star anise oil, or mixtures of menthol and cherry flavor.

[0181] These aromatic or flavoring substances may be present in an amount ranging from 0.0001 to 10% by weight (particularly in the composition) of the total composition, preferably from 0.001 to 6% by weight, more preferably from 0.001 to 4% by weight, most preferably from 0.01 to 1% by weight. Depending on the application or on a single occasion, it may be advantageous to use different amounts.

[0182] According to the present invention, all of the above-mentioned excipients and classes of excipients can be used without limitation, 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 not established.

[0183] In another aspect of the invention, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in a formulation for oral administration in the prevention or treatment of coronavirus infections.

[0184] Pharmaceutical formulations suitable for oral dosage form of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention may be administered as discrete units such as tablets, soft gelatin capsules, hard gelatin capsules, dragees or pills; powders or granules; solutions, syrups, drops, teas, solutions or suspensions in aqueous or non-aqueous liquids; edible foams or mousses; or oil-in-water or water-in-oil emulsions.

[0185] Thus, in oral dosage forms such as tablets or capsules, the active agent can be combined with a non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol or water. Powders are produced by grinding the compound to a suitable small particle size and mixing them with a pharmaceutical carrier, such as an edible carbohydrate, for example, starch or mannitol, in a similar manner. Flavoring agents, preservatives, dispersing agents or coloring agents can also be present.

[0186] Tablets are formulated by producing, granulating, or dry-pressing a powder mixture, adding a lubricant and disintegrant, and pressing the mixture into tablets. The powder mixture is produced by mixing the appropriately comminuted compound with a diluent or base as described above, and, if applicable, a binder such as carboxymethylcellulose, alginate, gelatin, or polyvinylpyrrolidone, a solution retarder such as paraffin, an absorption accelerator such as a quaternary salt, and / or an absorbent such as bentonite, kaolin, or dicalcium phosphate. The powder mixture can be granulated by wetting it with a binder such as syrup, starch paste, acacia mucilage, or a solution of cellulose or polymeric materials and pressing it through a sieve. As an alternative to granulation, the powder mixture can be passed through a tablet press to obtain lumps of uneven shape that are broken down to form granules. The granules can be lubricated with the addition of stearic acid, a stearate salt, talc, or mineral oil to prevent adhesion to the tablet die. The lubricated mixture is then pressed to produce tablets. The compounds according to the invention can also be combined with a free-flowing inert excipient and then pressed directly to give tablets without carrying out the granulation or dry-pressing step.

[0187] In another embodiment of the present invention, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts is provided in a hard gelatin capsule. These are produced by producing a powder mixture as described above and filling a formed gelatin capsule with it. Glazing agents and lubricants such as highly dispersed silica, talcum, magnesium stearate, calcium stearate, or polyethylene glycol can be added as solids to the powder mixture. Disintegrants or solubilizers such as agar, calcium carbonate, or sodium carbonate can also be added to improve the availability of the drug after ingestion of the capsule. Furthermore, suitable binders and / or coloring agents can be added to the mixture, if desired or necessary.

[0188] In another embodiment of the present invention, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts is contained in a soft gelatin capsule (SGC). SGCs dissolve as they pass through the digestive tract. They consist primarily of gelatin enriched with varying amounts of plasticizers, such as glycerol or sorbitan. The release rate depends on the specific formulation of the SGC carrier material. They are also suitable for sustained release of active agents. SGCs are particularly useful for administering poorly water-soluble active agents.

[0189] In another aspect of the invention, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts is contained in a chewable tablet or hard caramel, wherein the substance is incorporated into the matrix of the tablet or caramel.

[0190] In another aspect of the invention, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections in a formulation for inhaled administration.

[0191] For effective prophylactic or therapeutic treatment of coronavirus infections, which can cause pneumonia, pulmonary edema, and / or acute lung injury, it is advantageous for the 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, the composition according to the present invention, or the combination according to the present invention to reach the patient's alveoli. Therefore, the particle size must be small enough to reach the lowest part of the airways in the lung tissue. The best class of inhalation devices for inhalation administration of pharmaceutically active agents are so-called mesh nebulizers. 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 specialty purposes to sophisticated high-end mesh nebulizers for clinical or home treatment of serious diseases or conditions of the lower respiratory tract.

[0192] Suitable commercially available mesh nebulizers, jet nebulizers, ultrasonic nebulizers, dry powder inhalers and (pressurized) metered dose inhalers are PARI eFlow® rapid, PARI LC STAR®, PARI Velox and PARI Velox Junior (PARI GmbH, Sternberg, 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 inhaler MN-300 / 8, M-Neb Flow+ and M-neb® mesh nebulizer MN-300 / X (NEBU-TEC, Eisenfeld, 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, South 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® 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® (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® 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).

[0193] Mesh nebulizers with piezoelectric activation of the nebulization process, respectively vibrating mesh nebulizers, are preferred.

[0194] Thus, in another aspect of the invention, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections in a formulation for inhaled administration, wherein the inhaled administration is carried out by means of a vibrating mesh nebulizer.

[0195] 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 release nebulized aerosol continuously 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 lower respiratory tract than continuous-mode devices. The latter loses a larger volume of active-agent-containing aerosol to either the periphery or the upper respiratory tract, since aerosol release is not coupled to the respiratory cycle.

[0196] Therefore, trigger-activated mesh nebulizers, especially vibrating mesh nebulizers, are preferred.

[0197] Particularly preferred are trigger-activated mesh nebulizers with piezoelectric activation of the nebulization process.

[0198] 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.

[0199] The most preferred vibrating mesh nebulizer models are high-end models such as PARI eFlow® rapid, PARI Velox, Philips Respironics I-neb, M-neb® dose + mesh nebulizer inhaler MN-300 / 8, Aerogen® Solo, Vectura Fox, and Vectura Bayer Breelib™.

[0200] The mean droplet size is usually characterized as MMAD (mass median aerodynamic diameter). The size of individual droplets is called MAD (mass aerodynamic diameter). This value indicates whether 50% of the aerosol particles (droplets) are smaller or larger in diameter, respectively. 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 >5 μm and <10 μm usually reach the bronchi but not the alveoli. Particles with an MMAD between 100 nm and 1 μm do not deposit in the alveoli and are immediately exhaled. Therefore, the optimal range is an MMAD between 1 μm and 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).

[0201] A further commonly accepted quality parameter is the percentage of particles in the generated aerosol with diameters 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 diameters in the range <1 µm from the total percentage of particles in the generated aerosol with diameters in the range <5 µm (FPF; Fine Particle Fraction).

[0202] In another aspect of the present invention, the present application also provides a method for producing an aerosol according to the present invention for preventing or treating a coronavirus infection, comprising the following steps: a) filling a nebulization 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, a composition according to the present invention or a combination according to the present invention 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 nebulization chamber.

[0203] 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 Edition).

[0204] Therefore, the aforementioned method is also disclosed along with the vibration frequency range.

[0205] 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, the composition according to the invention or the combination according to the invention contained in said aqueous solution is sprayed into the aerosol generated.

[0206] The method of the present invention is particularly effective in nebulizing a high percentage of a pharmaceutically active agent from a provided aqueous solution within a short period of time, which is an important feature for patient compliance. A significant percentage of the patient population finds the inhalation process unpleasant, tiring, and physically demanding. However, active patient cooperation is essential for effective and targeted inhalation administration. Therefore, it is desirable to apply a therapeutically sufficient amount within the shortest possible period of time. Surprisingly, it has been shown that 95% of the substance provided in the aqueous solution can be nebulized within a 3-minute time span, which is an ideal time span for high patient compliance.

[0207] 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 generated aerosol is produced within 3 minutes after the start of nebulization with the mesh nebulizer.

[0208] While pharmaceutical active agents are typically provided in a single-dose container for each nebulization procedure, nebulizers and / or mouthpieces can be used for a certain period of time and need to be replaced at regular intervals. Cleaning of the nebulizer and mouthpiece is generally recommended after each nebulization. However, this practice does not guarantee reasonable patient compliance. However, even after careful cleaning, some aerosol deposits always remain within the nebulization chamber, outlet, and / or mouthpiece. Because aerosols are produced from aqueous solutions, these deposits pose a risk of generating bacterial bioburden, which could contaminate the inhaled aerosol. Deposits may also block the pores in the mesh membrane of mesh nebulizers. Generally, nebulizers and / or mouthpieces should be replaced every one or two weeks. Therefore, it is convenient to provide a combined product that includes the pharmaceutical and nebulizer.

[0209] Therefore, in another aspect of the present invention, the present application also refers to a kit comprising a mesh nebulizer and a pharmaceutically acceptable container containing an aqueous solution containing an effective amount of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts for preventing or treating a coronavirus infection, a composition according to the present invention or a combination according to the present invention, and optionally at least one pharmaceutically acceptable excipient.

[0210] In an alternative kit, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, the composition according to the present invention, or the combination according to the present invention is provided not in the form of an aqueous solution, but 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 the mesh nebulizer. These two containers can be completely separate containers, such as two vials, or, for example, a dual-chamber vial. For example, to dissolve the active agent, the membrane between the two chambers is pierced, allowing the contents of both chambers to mix.

[0211] Accordingly, the present application also discloses a kit comprising a mesh nebulizer, a first pharmaceutically acceptable container containing water for injection or saline, and a second pharmaceutically acceptable container containing an effective dosage of a solid form of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention, or a combination according to the invention for preventing or treating a coronavirus infection, and optionally at least one pharmaceutically acceptable excipient contained in the first pharmaceutically acceptable container and / or the second pharmaceutically acceptable container.

[0212] The aerosols produced by the methods according to the invention are each administered and self-administered by means of a mouthpiece, which optionally may further be included in the aforementioned kit.

[0213] A typical method involves transferring the provided or final aqueous solution to the nebulizer chamber using a syringe equipped with a needle. The aqueous solution is first drawn up into the syringe and then injected into the nebulizer chamber. Optionally, such a syringe and / or needle can be further included in the kit. Typical syringes made of, but not limited to, polyethylene, polypropylene, or cyclic olefin copolymers can be used, and typical gauges for stainless steel needles range from 14 to 27.

[0214] In yet another aspect of the present invention, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the present invention, or a combination according to the present invention is used for the prevention or treatment of coronavirus infection. The substance, composition, or combination is provided in the form of assisted ventilation, as an additive to the ventilation air of a cardiopulmonary bypass machine. As the condition of patients in intensive care units deteriorates, they often need to be ventilated indefinitely in such machines until their own breathing provides sufficient oxygen. Good results have been achieved using a metered-dose inhaler aerosol in combination with a Y-piece inhalation chamber. This allows for a 1.5- to 4-fold increase in the optimal bronchodilator dose (Fuller et al. (1994) Chest 105:214-218). 38% of the pharmaceutically active agent can be delivered (Marik et al. (1999) Chest 115:1653-1657). Alternatively, a constant-output mesh nebulizer, as evaluated in a scintigraphy study (Dugernier et al. (2016) Ann Intensive Care 6:73), resulted in a 10–15% rate. Vibrating mesh nebulizers provided better results than ultrasonic or jet nebulizers for antibiotic administration. When a constant-output vibrating mesh nebulizer was placed 10 cm above the inspiratory limb in a Y-shaped configuration and specific ventilation parameters (tidal volume 8 ml / kg, respiratory rate 12 c / min, duty cycle 50%, constant and low inspiratory flow rate less than 30 L / min, and end of inspiratory pause 20%) were set, 63% of the administered medication (ceftazidime, amikacin) reached the endotracheal tube entrance, compared with 37% extrapulmonary deposition (Lu et al. (2011) Am J Respir Crit Care Med 184:106–115). In most cases, the administered medication was distributed evenly between the two lungs. In pigs, the use of helium (He / O2) instead of nitrogen (N2 / O2) in the inhaled gas was found to increase ceftazidime concentrations in subpleural lung specimens (Tonnelier et al. (2005) Anesthesiology 102:995-1000).

[0215] In these cases, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts can be added to the intubation ventilation air in solid form (dry powder) or liquid form (in an aqueous solution or as a nebulized aerosol, as previously described).

[0216] Therefore, the present application also discloses 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections, wherein said substance, composition or combination is added to the ventilation air of a cardiopulmonary bypass machine.

[0217] In yet another aspect of the invention, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections, wherein said substance, composition or combination is applied in the form of a liposome, a micelle, a multilamellar vesicle or a cyclodextrin complex.

[0218] In yet another aspect of the invention, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections in a sublingual tablet formulation.

[0219] In yet another aspect of the invention, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections in a liquid dosage form.

[0220] The present application also discloses the parenteral administration of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, the composition according to the invention or the combination according to the invention in the prevention or treatment of coronavirus infections in the form of intravenous, intraarterial or intraperitoneal injection.

[0221] These liquid dosage forms include solutions, suspensions, and emulsions. Examples are water and water / propylene glycol solutions for parenteral injection or addition of sweeteners or opacifiers for oral solutions, suspensions and emulsions.

[0222] These liquid dosage forms can be stored in vials, IV bags, ampoules, cartridges, and pre-filled syringes. Suitable excipients include solubilizers, stabilizers, buffers, tonicity adjusters, bulking agents, thickening / reducing agents, surfactants, chelating agents, and adjuvants.

[0223] In yet another aspect of the present invention, 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the present invention or a combination according to the present invention for use in the prophylaxis or treatment of coronavirus infections, said substance, composition or combination is formulated as a lyophilisate, which can be reconstituted with water for injection or saline or a water / ethanol solution and then administered by injection.

[0224] Typical applications for intravenous injection include infusion pumps, hypodermic needles, drip chambers, peripheral cannulas (peripheral intravenous catheters), and pressure bags.

[0225] Generally, aqueous or saline solutions are preferred. In the case of poorly soluble pharmaceuticals according to the invention, ethanol or ethanol / water mixtures can also be used.

[0226] Further suitable liquid dosage forms include drops, eye drops, and ear drops.

[0227] While SARS-CoV and MERS-CoV primarily infect the lower respiratory tract, SARS-CoV-2 initially infects the pharynx / throat area. A small percentage of these patients subsequently develop pulmonary infection and pneumonia. These pharyngeal infections usually cause only mild or no cold-like symptoms, but these patients are highly contagious to their environment. In most cases, they are unaware that they have become spreaders of the infection. Therefore, there is a medical need to treat coronavirus infections while they are still in the pharyngeal stage, not only to treat such patients but also for epidemiological reasons to prevent the spread of the epidemic. For patients with pharyngeal infections, systemic administration routes, such as intravenous or oral administration, are not ideal, as highly effective drugs or drug combinations can also cause adverse side effects. Therefore, it is desirable to provide an administration route that locally treats infected pharyngeal tissue.

[0228] Therefore, in yet another aspect of the invention, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections in a formulation for pharyngeal administration.

[0229] Administration of the medication to the pharynx can be by suitable liquid dosage form as drops, lotion or tincture, or by viscous dosage form such as gel or hydrogel, by brushing the pharynx / pharyngeal area, gargling with a mouthwash, topical administration such as sublingual tablet, lozenge, throat spray or posterior pharyngeal wall injection.

[0230] Lotions are low-viscosity topical preparations intended for application to the skin or mucous membranes. Lotions are applied to the skin or mucous membranes with bare hands, a brush, a clean cloth, or cotton.

[0231] The advantage of lotions is that they can be spread thinly to cover large areas of skin or mucous membranes. Typical drugs that can be administered in lotion form include antibiotics, antiseptics, antifungals, corticosteroids, anti-acne agents, soothing agents, smoothing agents, moisturizing or protective agents, or anti-allergens.

[0232] Most lotions are oil-in-water emulsions that use substances such as cetearyl alcohol to hold the emulsion together, but water-in-oil lotions are also formulated. The key components are the aqueous and oily phases, the emulsion preventing separation of these two phases and the active pharmaceutical ingredient. A wide variety of excipients, such as fragrances, glycerol, petrolatum, dyes, preservatives, proteins, and stabilizers, are commonly added to lotions.

[0233] The thickness, consistency, and viscosity of the lotion can be adjusted during production. Producing a lotion can be done in two steps: a) dispersing emollients and lubricants in the oil phase along with blending agents and thickeners, and b) dispersing fragrances, colorants, and preservatives in the aqueous phase. Pharmaceutically active ingredients are dissolved in both steps depending on the ingredients involved and the desired properties of the lotion.

[0234] Tinctures are typically alcoholic extracts or formulations. Solvent concentrations of 25-60% (or even 90%) are common. Other solvents for producing tinctures include vinegar, glycerin, diethyl ether, and propylene glycol. Ethanol has the advantage of being a good solvent for both acidic and alkaline ingredients. Tinctures using glycerin are called glycerites. Glycerin is generally a poorer solvent than ethanol. Vinegar, being acidic, is a better solvent for obtaining alkaloids but a poor solvent for acidic ingredients.

[0235] Gels are colloids in which a solid dispersed phase forms a network in combination with a fluid continuous phase, resulting in a viscous, semi-rigid sol. Gel properties range from soft and weak to hard and tough. They are defined as substantially dilute, cross-linked systems that do not exhibit flow at steady state. By weight, gels are mostly liquid, but behave like solids due to a three-dimensional cross-linked network within the liquid. It is the cross-links within the fluid that give the gel its consistency and contribute to its adhesive stickiness. A gel is a dispersion of liquid molecules within a solid medium.

[0236] Hydrogels are networks of hydrophilic polymer chains, sometimes found as colloidal gels in which water is the dispersion medium. The three-dimensional solid results from hydrophilic polymer chains held together by crosslinks. Due to the inherent crosslinks, the structural integrity of the hydrogel network prevents it from dissolving in high concentrations of water. Hydrogels are natural or synthetic polymer networks that are highly absorbent (they can contain over 90% water). Due to their significant water content, hydrogels also possess a degree of flexibility very similar to that of natural tissue. In medicine, hydrogels can encapsulate chemical systems that, when stimulated by external factors such as pH changes, can release certain pharmaceutically active agents into the environment, most often through a gel-sol transition to a liquid state.

[0237] Suitable gel-forming agents may be selected from the group including, but not limited to, agar, algin, alginic acid, bentonite, carbomer, carrageenan, hectorite, hydroxyethyl cellulose, hydroxypropyl cellulose, polyvinyl alcohol, polyvinylpyrrolidone, sodium carbomer.

[0238] A mouthwash is a liquid that is passively held in the mouth or rinsed around the mouth by contraction of the muscles around the mouth and / or movement of the head, and may be gargled, with the head tilted back and the liquid foaming in the back of the mouth. Thus, an aqueous or alcoholic solution of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention can be formulated and administered to the pharynx.

[0239] Sublingual dosage forms can bypass hepatic metabolism, making them an alternative to oral drug delivery. Rapid onset of pharmacological effects is often desired for some drugs, especially those used to treat acute disorders. Sublingual tablets disintegrate quickly, and the small amount of saliva present is usually sufficient to achieve disintegration of the dosage form, which results in better dissolution and increased bioavailability.

[0240] Drugs must be sufficiently lipophilic to be able to partition through the lipid bilayer, but not so lipophilic that they cannot redistribute once inside. According to the diffusion model of absorption, the flux across the lipid bilayer is directly proportional to the concentration gradient. Therefore, lower salivary solubility results in a lower absorption rate, and vice versa. In general, drugs formulated for sublingual use should ideally have a molecular weight below 500 to facilitate their diffusion. The oral cavity has a narrow pH range, between 5.0 and 7.0. The inclusion of a suitable buffer in the formulation of ionizable drugs allows for control of the pH of aqueous saliva.

[0241] To avoid the possibility of unpleasant taste or odor, drug taste masking is necessary. Sweeteners, flavoring agents, and other flavoring agents are essential ingredients. Sugar-based excipients dissolve rapidly in saliva, resulting in endothermic dissolution. They create a pleasant sensation in the mouth and are best suited for sublingual tablets along with other flavors.

[0242] Typical techniques for manufacturing sublingual tablets include direct compression, compression molding, freeze-drying, and hot-melt extrusion (Khan et al. (2017) J Pharmaceut Res 16:257-267).

[0243] If swallowing is avoided, administration of the pharmaceutically active agent via a sublingual tablet can also reach the pharynx / throat locally. Absorption of the pharmaceutically active agent occurs primarily via the pharyngeal mucosa.

[0244] A lozenge (troche) is a small disk or diamond-shaped object composed of a solidified paste containing an astringent, antiseptic, or thinning agent used for the local treatment of the mouth or throat; the lozenge is held in the mouth until it dissolves. The vehicle or base of a lozenge is usually sugar, an adhesive made by mixing with acacia or tragacanth, a fruit paste made from black or red currants, rose confectionery, or balsam of Tolu.

[0245] In particular, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections in a formulation for pharyngeal administration, wherein the pharyngeal administration is carried out by throat spray.

[0246] A throat spray is a medicated liquid that is typically administered to the throat as a spray to treat a sore throat or cough.

[0247] Throat sprays typically contain local anesthetics (e.g., lidocaine, benzocaine), antiseptics (e.g., chlorhexidine, cetylpyridinium chloride), herbal extracts, or a combination thereof. Whatever the formulation, it should not contain excessive amounts of sugar or ethanol, which further irritate the mucous membrane. And finally, the user should not experience any unpleasant aftertaste.

[0248] The current standard for throat sprays is a metering pump attached to a bottle containing 10-30 ml of liquid formulation. The formulation is filled into a glass or plastic bottle with the pump secured by a screw closure, either crimped onto the bottleneck, or simply snapped on. Regardless of the securing option chosen, the system must be tight and no leaks are observed during transport or handling by the user. The container is usually made from glass or plastic.

[0249] Typically, throat spray pumps deliver doses ranging from 50 to 200 μl per actuation. For targeted administration, pumps are equipped with actuators with extended nozzles. Nozzle lengths may range from 30 mm to 70 mm. While it is easier to target the affected area with such long, fixed nozzles, they may be too large for the user to carry, which is why actuators with folding or swiveling nozzles are preferred.

[0250] Alternatively, the device utilizes a continuous valve. Because the formulation is aerosolized while the actuator is depressed, the continuous valve delivers targeted therapy but not precise dosing. One technical solution is a tin or aluminum can with a pressurized headspace. When the valve is actuated, the increased internal pressure forces the formulation out of the can as long as the valve system is depressed.

[0251] A related but more sophisticated system is the bag-on-valve (BOV) system. The product is placed inside the bag while the propellant (most often compressed air) fills the space between the bag and the outer canister. When a continuous valve is activated, the product is squeezed out of the bag by the compressed air. The BOV system functions in any 360° orientation.

[0252] Care should be taken with throat spray formulations, as they may contain ingredients that are highly aggressive and can reduce surface tension. A simple test of spray performance will ensure that the formulation can be aerosolized by the system and that the spray pattern and particle size delivered are appropriate for the intended use.

[0253] The spray pattern and droplet size distribution are the most important parameters of throat spray. Spray pattern is a term used to describe the spray angle and plume shape of a fully developed spray. Once the spray is fully developed using laser diffraction techniques, the droplet size is characterized. Fine particles (droplets with a mean dynamic diameter less than 10 μm) should be as low as possible to avoid droplet deposition in the lower respiratory tract.

[0254] Recently, several carragelose-based throat sprays have emerged that claim to protect against upper respiratory tract infections of viral origin. The first polymer in this platform is Carragelose®, a broadly active antiviral compound for treating respiratory diseases. In addition to its moistening effect, the compound prevents viral binding to mucosal cells.

[0255] Alternatively, a handheld nebulizer with a high output rate and adjusted droplet size can be used for deposition in the upper respiratory tract. Breathing through a face mask can result in droplet deposition on mucous membranes throughout the upper respiratory tract (see Marx and Nadler (2018) Drug Development & Delivery).

[0256] In particular, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of coronavirus infections in a formulation for pharyngeal administration, wherein the pharyngeal administration is carried out by posterior pharyngeal wall injection.

[0257] This technique is used in orthopedic surgery for pharyngoplasty and other procedures using calcium hydroxide apatite injections. However, local injections into pharyngeal tissue can also be used to administer pharmaceutically active agents. The injection solution can be similar to that used for intravenous or intramuscular injections. Aqueous solutions, saline, or, for fairly lipophilic pharmaceutically active agents, an ethanol / water mixture is preferred.

[0258] In a further aspect of the invention, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in the prevention or treatment of SARS-CoV-2 infection in a formulation for nasal administration.

[0259] In particular, nasal administration is by means of nasal sprays or nasal drops.

[0260] Common formulation types used in nasal spray products are solutions, suspensions, and emulsions. Nasal spray formulations can be aqueous, hydroalcoholic, or non-aqueous-based. Depending on the type of system, the formulation contains a range of functional excipients, including solvents and cosolvents; mucoadhesives; pH buffers; antioxidants; preservatives; osmolality and tonicity agents; penetration enhancers; suspending agents; and surfactants. The formulation type and the excipients selected are determined by the solubility and stability of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, as well as the concentration required to deliver an effective dose in a typical 100 μl spray (see Kulkarni and Shaw (2016) in: Essential Chemistry for Formulators of Semisolid and Liquid Dosages, Elsevier). The aforementioned Carragelose® technology is also used in nasal sprays.

[0261] Nose drops are administered in a similar formulation, but are dispensed by dripping instead of pressing a dispenser.

[0262] In particular, the application relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the invention or a combination according to the invention for use in a formulation in the prevention or treatment of SARS-CoV-2 infection for nasal administration, wherein nasal administration is carried out by nasal spray or nasal drops.

[0263] The mucous membranes of the eyes are known to be another entry point for SARS-CoV-2 into the organism, and people can get the virus on their hands while rubbing their eyes, for example.

[0264] The present application therefore also relates to 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, the composition according to the invention or the combination according to the invention, wherein the administration of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, the composition according to the invention or the combination according to the invention is carried out by means of eye drops.

[0265] Eye drops are primarily aqueous solutions containing pharmaceutical active ingredients. The pH is usually adjusted to 7.1–7.5. Common buffers for eye drops are boric acid and monobasic sodium phosphate. Tonicity should be adjusted to an osmolality isotonic with the corneal epithelium (225–430 mosm / kg) using 0.9% saline (or another tonicity agent, such as potassium nitrate, boric acid, sodium acetate, sodium acetate phosphate buffer, or mannitol). Suitable preservatives include thiomersal, organic mercury compounds such as phenylmercury, benzalkonium chloride, chlorhexidine, and benzyl alcohol. To extend contact time, contact-time-increasing thickening agents (thickeners) such as cellulose derivatives (hypromellose, methylcellulose, hydroxypropylmethylcellulose), hyaluronic acid, cellulose acetate phthalate, polyethylene glycol, polyvinyl alcohol, or poloxamer can be added. Wetting agents or surfactants such as benzalkonium chloride, polysorbate 20, polysorbate 80, dioctyl sodium sulfosuccinate, etc. may be included. Some amino acids, alone or in combination with sodium hyaluronate, may help promote tissue reconstitution as needed. Suitable amino acids are glycine, leucine, lysine, and proline (see EP 1940381).

[0266] In a further aspect of the present invention, a method for treating a coronavirus infection is disclosed, wherein an effective amount of 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a composition according to the present invention or a combination according to the present invention is administered to a patient in need thereof or to a healthy subject at risk of being infected with coronavirus. [Example]

[0267] In all experiments, solutions of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt were prepared using the anhydrous Form I of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt described above (provided by MetrioPharm).

[0268] Example 1: 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt inhibits replication of SARS-CoV-2 in infected VeroB4 cells

[0269] Western blot (WB) analysis was performed to investigate whether 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt affected the spread of viral infection. Vero B4 cells (National Institutes of Health, Bethesda, MD, USA; Meyer et al. (2015) Emerg Infect Dis 21:181-182) were maintained in Dulbecco's modified Eagle's medium (DMEM) containing 10% (v / v) inactivated fetal calf serum (FCS), 2 mM L-glutamine, 100 U / mL penicillin, and 100 μg / mL streptomycin. Confluent monolayers of Vero B4 cells were incubated with a 1:100 dilution of wild-type SARS-CoV-2 virus. PR-1 Cells (isolated from a 61-year-old patient 6 days after the estimated date of infection and 2 days after the onset of mild COVID-19 symptoms) were infected for 1 hour in FCS-free DMEM containing 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt. Cells were then washed with PBS (phosphate-buffered saline) and provided with fresh medium containing noncytotoxic concentrations (0.5 μM, 1 μM, 2 μM) of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt. Treatment with 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt was performed throughout the experimental procedure. Virus-containing cell culture supernatant was collected 3 days postinfection (dpi). Virions were purified from the cell culture supernatant using 20% ​​(w / v) sucrose buffer (20,000 × g, 4°C, 90 min). Cells were washed with PBS, and the pellet was dissolved in SDS (sodium dodecyl sulfate) sample buffer. Cells were separated by SDS-PAGE gel electrophoresis, transferred to nitrocellulose membranes, blocked with 3% bovine serum albumin, and incubated with the appropriate primary antibodies. SARS-CoV-2 proteins were visualized using serum from convalescent SARS-CoV-2 patients. Anti-human secondary antibodies conjugated to horseradish peroxidase were obtained from Dianova (Hamburg, Germany). Visualization was performed by electrochemiluminescence.

[0270] Here, we demonstrated inhibition of SARS-CoV-2 replication in Vero B4 cells. 5-Amino-2,3-dihydro-1,4-phthalazinedione sodium salt showed a clear reduction in SARS-CoV-2 nucleocapsid protein. The respective gel bands are shown in Figure 1A.

[0271] Densitometric evaluation of SARS-CoV-2 nucleocapsids was performed using the analysis program AIDA®. Densitometric evaluation allows for quantification of signal intensity in Western blots and therefore allows conclusions regarding the amount of a specific protein in the sample. The evaluation clearly showed that after the addition of 5-amino 2,3-dihydro-1,4-phthalazinedione sodium salt, SARS-CoV-2 protein production was inhibited in a dose-dependent manner (Figure 1B). Statistical analysis was performed using an unpaired t-test with Welch's correction; **p<0.01, *p<0.05.

[0272] Example 2: At effective concentrations, 5-amino 2,3-dihydro-1,4-phthalazinedione sodium salt is not cytotoxic in VeroB4 cell cultures

[0273] To address the question of whether 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt exhibits cytotoxic effects in the above system, uninfected Vero B4 cells were treated with increasing concentrations of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt (0.25 mM, 0.5 mM, 1 mM, 2 mM, and 4 mM) in parallel with Western blot studies. Toxicity was assessed with a WST (water-soluble tetrazolium salt 1) assay. Here, viable cells with an intact mitochondrial succinate-tetrazolium dehydrogenase system result in the enzymatic conversion of the faintly red tetrazolium salt WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzenedisulfonate) to dark red formazan. This color change can be measured photometrically in a spectrophotometer. Therefore, the WST assay is a very sensitive method for measuring the toxicity of substances to cellular metabolism. The value for untreated cells was set to 100%.

[0274] It can be shown that 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt does not exhibit any significant toxic effects at antivirally effective concentrations in Vero B4 cells during an observation period of 3 days.

[0275] In Figure 2, the percentage of viable cells is shown compared to untreated cells. The value for untreated cells was set to 100%. 1 μM staurosporine (an indolocarbazole compound derived from Streptomyces staurosporeus, an apoptosis inducer) was used as a positive control. Statistical analysis was performed using an unpaired t-test with Welch's correction; **p<0.01, *p<0.05.

[0276] Therefore, it can be said that the antiviral effect of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt is not due to a nonspecific cytotoxic effect. [Brief explanation of the drawings]

[0277] [Figure 1A] Figure 1 shows Western blot bands of SARS-CoV-2 nucleocapsid after 3 days of treatment with 0.5 mM, 1 mM and 2 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt relative to vehicle and untreated cells, respectively. [Figure 1B] Figure 1 shows densitometric assessment of SARS-CoV-2 nucleocapsid detected in Western blot bands after 3 days of treatment with 0.5 mM, 1 mM, and 2 mM 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt relative to vehicle and untreated cells, respectively. The percentage of viral protein detected is shown. Vehicle was set to 100%. (Mean ± SEM; n=3 / group; each duplicate; **p<0.01, *p<0.05) [Figure 2] Figure 1 shows cell viability in WST assay after treatment with different concentrations of 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt for 3 days. The percentage of viable cells is shown. Untreated cells were set as 100%. Staurosporine (1 μM) was used as a positive control. (Mean ± SEM; n = 3 / group; each replicate)

Claims

1. A preventive or therapeutic agent for coronavirus infection by inhibiting coronavirus replication, comprising 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts.

2. A preventive or therapeutic agent for coronavirus infection according to claim 1, The pharmaceutically acceptable salt of 5-amino-2,3-dihydro-1,4-phthalazinedione is 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt. A preventive or therapeutic agent for coronavirus infections.

3. A preventive or therapeutic agent for coronavirus infection according to claim 2, The 5-amino-2,3-dihydro-1,4-phthalazinedione sodium salt is provided as one of crystalline anhydrous polymorphic Forms I, II, or III characterized by the following crystallographic values ​​as determined by 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 2-theta 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 2-theta 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 2-theta 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 A preventive or therapeutic agent for coronavirus infection.

4. A preventive or therapeutic agent for coronavirus infection according to any one of claims 1 to 3, The coronavirus infection is selected from the group consisting of SARS-CoV, MERS-CoV, SARS-CoV-2, HCoV-HKU1, HCoV-NL-63, HCoV-OC43 and HCoV-229E infections; A preventive or therapeutic agent for coronavirus infections.

5. A pharmaceutical composition for use in the prevention or treatment of coronavirus infection by inhibiting coronavirus replication, comprising: A pharmaceutical composition containing 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts, a carrier, and at least one pharmaceutically acceptable excipient. Pharmaceutical compositions.

6. A pharmaceutical composition for use in the prevention or treatment of coronavirus infection by inhibiting coronavirus replication, comprising: 5-amino-2,3-dihydro-1,4-phthalazinedione or one of its pharmaceutically acceptable salts; steroidal and non-steroidal anti-inflammatory drugs, immunomodulators, immunosuppressants; anti-infectives such as antibiotics, antiretrovirals, antivirals, antifungals and antiprotozoal agents, analgesics, anticoagulants, antiplatelet agents, bronchodilators, pulmonary vasodilators, mucolytics, pulmonary surfactants, antioxidants, ENaC activators, HMG-CoA reductase inhibitors or AT １ and at least one active agent selected from the group comprising receptor antagonists.

7. A preventive or therapeutic agent for coronavirus infection according to any one of claims 1 to 3, or a pharmaceutical composition according to any one of claims 4 to 6, orally applied in the form of tablets, soft gelatin capsules, hard gelatin capsules, dragees, pills, powders, granules, liquids, syrups, drops, teas, solutions or suspensions in aqueous or non-aqueous liquids, edible foams, mousses, oil-in-water emulsions or water-in-oil emulsions; A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection.

8. A preventive or therapeutic agent for coronavirus infection according to any one of claims 1 to 3, or a pharmaceutical composition according to any one of claims 4 to 6, A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection, which is a formulation for inhalation administration.

9. A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection according to claim 8, A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection, wherein the inhalation administration is performed using a vibrating mesh nebulizer.

10. A preventive or therapeutic agent for coronavirus infection according to any one of claims 1 to 3, or a pharmaceutical composition according to any one of claims 4 to 6, added to the ventilation air of cardiopulmonary bypass machines, A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection.

11. A preventive or therapeutic agent for coronavirus infection according to any one of claims 1 to 3, or a pharmaceutical composition according to any one of claims 4 to 6, A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection, which is a sublingual tablet formulation.

12. A preventive or therapeutic agent for coronavirus infection according to any one of claims 1 to 3, or a pharmaceutical composition according to any one of claims 4 to 6, A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection, wherein the preventive or therapeutic agent or pharmaceutical composition for coronavirus infection is administered by nasal spray, nasal drops, or eye drops.

13. A preventive or therapeutic agent for coronavirus infection according to any one of claims 1 to 3, or a pharmaceutical composition according to any one of claims 4 to 6, A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection, which is a preparation for pharyngeal administration.

14. The preventive or therapeutic agent or pharmaceutical composition for coronavirus infection according to claim 13, The pharyngeal administration is performed by throat spray. A preventive or therapeutic agent or pharmaceutical composition for coronavirus infection.

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