Combination of elderberry extract and quinine for the prophylaxis and treatment of RNA virus infections

Abstract

The present application relates to a combination of elderberry extract and quinine for use in the prevention or treatment of RNA virus infections, in particular coronavirus and influenza virus infections. Pharmaceutic compositions and advantageous formulation techniques are disclosed. Moreover, the present invention also relates to anthocyanin compounds and phenolic compounds which may be obtained from elderberry extract, or other suitable sources, or synthesized de novo, and their respective use(s) in the prevention or treatment of RNA virus infections, in particular coronavirus infections. Moreover, the present invention also relates to elderberry extract, for use as a monotherapy in the prevention or treatment of influenza virus infections. Furthermore, the present invention also relates to quinine, for use as a monotherapy in the prevention or treatment of influenza virus infections. Furthermore, the present invention also relates to combinations of individual anthocyanin compounds or phenolic compounds with quinine, and to the use(s) of such combinations in the prevention or treatment of RNA virus infections, in particular coronavirus and influenza virus infections.

Description

[0001] Combination of elderberry extract and quinine for the prophylaxis and treatment of RNA virus infections

[0002] FIELD OF THE INVENTION

[0003] The present application relates to a combination of elderberry extract and quinine for use in the prevention or treatment of RNA virus infections, in particular coronavirus and influenza virus infections. Pharmaceutic compositions and advantageous formulation techniques are disclosed. Moreover, the present invention also relates to anthocyanin compounds and phenolic compounds which may be obtained from elderberry extract, or other suitable sources, or synthesized de novo, and their respective use(s) in the prevention or treatment of RNA virus infections, in particular coronavirus infections. Moreover, the present invention also relates to elderberry extract, for use as a monotherapy in the prevention or treatment of influenza virus infections. Furthermore, the present invention also relates to quinine, for use as a monotherapy in the prevention or treatment of influenza virus infections. Furthermore, the present invention also relates to combinations of individual anthocyanin compounds or phenolic compounds with quinine, and to the use(s) of such combinations in the prevention or treatment of RNA virus infections, in particular coronavirus and influenza virus infections.

[0004] BACKGROUND OF THE INVENTION

[0005] RNA viruses build a major group of viruses. They have ribonucleic acid (RNA) as their genetic material. Most RNA viruses possess a single-stranded RNA (ssRNA), but some are doublestranded (dsRNA). Many human-pathogenic viral infections are due to RNA viruses e.g., rhinitis, influenza, HRSV, SARS-CoV, SARS-CoV-2, MERS, Dengue fever, West Nile fever, Ebola, rabies, polio, mumps and measles.

[0006] These viral infections are either treated symptomatically or with a virostatic drug. Virostatics, however, display usually an efficacy against one or more specific virus families or species. Moreover, RNA viruses are known to have a high mutation rate (approximately 10'6to 10'4s / n / c) due mainly to their lack of exonuclease proofreading activity displayed by their RNA polymerases (cf. Steinhauer et al. (1992) Gene 122: 281-288). As most virostatic drugs act on a viral target they are at risk to lose their efficacy over time when the genetic drift of a virus starts to escape the actions of a virostatic. Therefore, it would be desirable to have a broad- spectrum anti-RNA-viral agent at hand that acts on targets of the host cells which are much less likely to undergo mutations.

[0007] End of 2019 beginning of 2020 the SARS-CoV-2 pandemic started to spread all over the world causing up to 20 million casualties (WHO estimate from May 2023). Before the development of anti-SARS-CoV-2 vaccines and the establishment of pharmaceutic treatment regimes, the infected patients with a severe, potentially life-threatening manifestation of COVID-19 could only be treated symptomatically. There was no effective antiviral drug available to tackle this new and overwhelming challenge of mankind. A favored hypothesis is that SARS-CoV-2 arose from a zoonosis. It is generally believed that such an event can occur any time again, confronting mankind with an uncontrollable novel virus. This may also occur by malicious mutations of hitherto harmless viruses, or even by gain-of-function modified viruses that would be released by accident or intentionally.

[0008] This situation increased the scientific awareness to develop medications or dietary supplements that may combat such novel virus threats immediately from the beginning of a pandemic, independently from the novel virus species. This goal can be mainly achieved by compounds that act on host cell targets.

[0009] Further, it would be desirable that such a drug can be given prophylactically. A prerequisite for a prophylactic use is that such a drug has no or only mild adverse side effects.

[0010] Surprisingly, this task is solved by the administration of elderberry extract and quinine. It was found that a combination of elderberry extract and quinine is able to modify gene expression in host cells in such a way that the inborn defense mechanism of the affected organism is boosted in such a way that the replication of virtually all RNA viruses can be effectively and specifically suppressed.

[0011] SUMMARY OF THE INVENTION

[0012] Thus, the present application discloses a combination of elderberry extract and quinine.

[0013] In one aspect of the invention, a combination of elderberry extract and quinine for use in medicine is disclosed.

[0014] In another aspect of the invention a combination of elderberry extract and quinine for use as a dietary supplement is disclosed.

[0015] The present disclosure refers thus to a combination of elderberry extract and quinine for use in the prophylaxis or treatment of an RNA virus infection. In a preferred embodiment the present disclosure refers thus to a combination of elderberry extract and quinine for use in the prophylaxis or treatment of a respiratory RNA virus infection.

[0016] In particular a combination of elderberry extract and quinine for use in the prophylaxis or treatment of a coronaviral infection is disclosed, in particular in the prophylaxis or treatment of a SARS-CoV-2 infection, respectively COVID-19.

[0017] In particular a combination of elderberry extract and quinine for use in the prophylaxis or treatment of an orthomyxovirus infection is disclosed, in particular in the prophylaxis or treatment of an influenza virus infection.

[0018] According to one aspect, the present disclosure relates to a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts.

[0019] According to a further aspect, the present disclosure relates to a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts, for use as a dietary supplement.

[0020] According to a further aspect, the present disclosure relates to a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts for use in the prophylaxis or treatment of an RNA viral infection.

[0021] In one embodiment of the aforementioned disclosure, the RNA viral infection is a coronaviral infection.

[0022] In a preferred embodiment, the coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV and HCoV-HKU1, HCoV-NL-63, HCoV-OC43 and HCoV-229E infection.

[0023] In another embodiment of the aforementioned disclosure, the RNA viral infection is an influenza viral infection.

[0024] In a preferred embodiment, the influenza viral infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0025] In another embodiment of the aforementioned disclosure, the RNA viral infection is selected from a group comprising infections with a virus from the families of orthomyxoviridae, picornaviridae, pneumoviridae, hepeviridae, reoviridae, togaviridae, flaviviridae, arenaviridae, filoviridae, hantaviridae and rhabdoviridae.

[0026] In a preferred embodiment, said viral infection is selected from a group comprising RVs, PV, HRSV, HMPV, HEV, RoV, RuV, TBEV, J EV, ZIKV, YFV, WNV, DENV, LASV, EBOV, MARV, HTNV, PUUV, DOBV, SEOV, SNV, ANDV and RABV infection.

[0027] In accordance with any of the aforementioned aspects and embodiments, said combination may further comprise at least one of aromatic and flavoring substances, sweeteners, pH- regulators, acidity regulators, preservatives, antioxidants, or thickening agents.

[0028] The present disclosure also relates to a method of prevention or treatment of an RNA viral infection, as defined herein, said method comprising administering a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts, to a patient in need thereof.

[0029] The present disclosure also relates to a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts, for the manufacture of a medicament for the prevention or treatment of an RNA viral infection, as defined herein.

[0030] In a further aspect, the present disclosure also relates to a composition containing the following ingredients: elderberry in a weight range of 0.01 wt.-% to 2 wt.-%; quinine or a water-soluble pharmaceutically acceptable salt thereof in a weight range of 0.01 wt.-% to 1 wt.-%; a pH regulator and / or acidity regulator in a weight range of 0.01 wt.-% to 2 wt.-%; a sweetener in a weight range of 1 wt.-% to 25 wt.-%; one or more aromatic and flavoring substances in a weight range of 0.01 wt.-% to 2 wt.-%; and water in a weight range of 1 .4 wt.-% to 68 wt.-%, wherein the relative weight ranges of all included components sum up to 100%.

[0031] In one embodiment, said composition further comprises at least one of preservatives, antioxidants, or thickening agents in a total range of 0 wt.-% to 5 wt.-% of the composition, wherein the relative weight ranges of all included components sum up to 100%. In a preferred embodiment, said composition contains the following ingredients: elderberry extract (0.06 wt.-%) a water-soluble pharmaceutically acceptable quinine salt (0.0104 wt.-%) anhydric citric acid (0.25 wt.-%) invert sugar syrup (13 wt.-%) flavoring agents (0.1 wt.-%), filled up to 100 wt.-% with water.

[0032] In a further aspect, the present disclosure also relates to a pharmaceutical composition for use in the prophylaxis or treatment of an RNA viral infection, wherein said composition contains a combination as defined herein, in particular a combination as described in the aforementioned aspects or embodiments, a carrier and at least one pharmaceutically acceptable excipient.

[0033] In one embodiment of this aspect, said pharmaceutical composition is intended for use in medicine.

[0034] In yet a further aspect, the present disclosure also relates to an isolated anthocyanin compound, for use in the prophylaxis or treatment of a coronaviral infection, wherein said isolated anthocyanin compound is selected from cyanidine-3-O-sambubioside chloride, cyanidine-3-glucoside and cyanidine-3-O-sambubioside-5-O-glucoside, wherein preferably, said isolated anthocyanin compound is selected from cyanidine-3-O-sambubioside chloride and cyanidine-3-glucoside.

[0035] In yet a further aspect, the present disclosure also relates to an isolated phenolic compound, for use in the prophylaxis or treatment of a coronaviral infection, wherein said isolated phenolic compound is selected from rutin, isoquercitrin, chlorogenic acid and neochlorogenic acid, wherein, preferably, said phenolic compound is selected from rutin and isoquercitrin.

[0036] With respect to the aforementioned aspects of said anthocyanin compound and said phenolic compound, it is preferred that the coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV and HCoV-HKU1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infection.

[0037] Preferably, said coronaviral infection is an infection by SARS-CoV-2. The present disclosure also relates to a method of prevention or treatment of a coronaviral infection, as defined herein, said method comprising administering an isolated anthocyanin compound, to a patient in need thereof, wherein said isolated anthocyanin compound is selected from cyanidine-3-O-sambubioside chloride, cyanidine-3-glucoside and cyanidine-3- O-sambubioside-5-O-glucoside, wherein preferably, said isolated anthocyanin compound is selected from cyanidine-3-O-sambubioside chloride and cyanidine-3-glucoside.

[0038] The present disclosure also relates to an isolated anthocyanin compound, as defined herein, for the manufacture of a medicament for the prevention or treatment of a coronaviral infection, as defined herein.

[0039] The present disclosure also relates to a method of prevention or treatment of a coronaviral infection, as defined herein, said method comprising administering an isolated phenolic compound, to a patient in need thereof, wherein said isolated phenolic compound is selected from rutin, isoquercitrin, chlorogenic acid and neochlorogenic acid, wherein, preferably, said isolated phenolic compound is selected from rutin and isoquercitrin.

[0040] The present disclosure also relates to an isolated phenolic compound, as defined herein, for the manufacture of a medicament for the prevention or treatment of a coronaviral infection, as defined herein.

[0041] In yet a further aspect, the present disclosure also relates to elderberry extract, for use in the prophylaxis or treatment of an influenza viral infection, wherein, preferably, said elderberry extract is used on its own without additional other active pharmaceutical ingredients.

[0042] The present disclosure also relates to a method of prevention or treatment of an influenza viral infection, as defined herein, said method comprising administering an elderberry extract on its own without additional other active pharmaceutical ingredients, to a patient in need thereof.

[0043] The present disclosure also relates to an elderberry extract on its own without additional other active pharmaceutical ingredients, for the manufacture of a medicament for the prevention or treatment of an influenza viral infection, as defined herein. In yet a further aspect, the present disclosure also relates to quinine or one of its pharmaceutically acceptable salts, for use in the prophylaxis or treatment of an influenza viral infection, wherein, preferably, said quinine or said pharmaceutically acceptable salt thereof, is used on its own without additional other active pharmaceutical ingredients.

[0044] The present disclosure also relates to a method of prevention or treatment of an influenza viral infection, as defined herein, said method comprising administering quinine, or one of its pharmaceutically acceptable salts, on its own without additional other active pharmaceutical ingredients, to a patient in need thereof.

[0045] The present disclosure also relates to quinine, or one of its pharmaceutically acceptable salts on its own without additional other active pharmaceutical ingredients, for the manufacture of a medicament for the prevention or treatment of an influenza viral infection, as defined herein.

[0046] With respect to the aforementioned aspects of said elderberry extract and said quinine, each being used on their own without additional other active pharmaceutical ingredients being present, it is preferred that the influenza viral infection is selected from a group consisting of IAV, IBV, ICV and IDV infection. Preferably, said influenza viral infection is an infection by influenza A virus (IAV).

[0047] The present disclosure furthermore relates to a combination of an individual anthocyanin compound and quinine, or one of its pharmaceutically acceptable salts, wherein the individual anthocyanin compound is selected from cyanidine-3-O-sambubioside chloride, cyanidine-3- glucoside and cyanidine-3-O-sambubioside-5-O-glucoside, wherein preferably, said individual anthocyanin compound is selected from cyanidine-3-O-sambubioside chloride and cyanidine- 3-glucoside.

[0048] The present disclosure furthermore relates to a combination as defined in the preceding paragraph, for use in the prophylaxis or treatment of an RNA viral infection.

[0049] In one embodiment, such RNA viral infection is either a coronoviral infection or an influenza viral infection. Preferably, the coronoviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV and HCoV-HKU1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infection. Equally preferably, the influenza viral infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0050] In another embodiment, said RNA viral infection is selected from a group comprising infections with a virus from the families of orthomyxoviridae, picornaviridae, pneumoviridae, hepeviridae, reoviridae, togaviridae, flaviviridae, arenaviridae, filoviridae, hantaviridae and rhabdoviridae. Preferably, said viral infection is selected from a group comprising RVs, PV, HRSV, HMPV, HEV, RoV, RuV, TBEV, JEV, ZIKV, YFV, WNV, DENV, LASV, EBOV, MARV, HTNV, PUUV, DOBV, SEOV, SNV, ANDV and RABV infection.

[0051] The present disclosure also relates to a method of prevention or treatment of an RNA viral infection, as defined herein, said method comprising administering a combination of an individual anthocyanin compound and quinine, or one of its pharmaceutically acceptable salts, to a patient in need thereof, wherein the individual anthocyanin compound is selected from cyanidine-3-O-sambubioside chloride, cyanidine-3-glucoside and cyanidine-3-O- sambubioside-5-O-glucoside, wherein preferably, said iindividual anthocyanin compound is selected from cyanidine-3-O-sambubioside chloride and cyanidine-3-glucoside.

[0052] The present disclosure also relates to a combination of an individual anthocyanin compound, as defined herein, and quinine, or one of its pharmaceutically acceptable salts, for the manufacture of a medicament for the prevention or treatment of an RNA viral infection, as defined herein

[0053] The present disclosure furthermore relates to a combination of an individual phenolic compound and quinine, or one of its pharmaceutically acceptable salts, wherein the individual phenolic compound is selected from rutin, isoquercitrin, chlorogenic acid and neochlorogenic acid, wherein, preferably, said individual phenolic compound is selected from rutin and isoquercitrin.

[0054] The present disclosure furthermore relates to a combination as defined in the preceding paragraph, for use in the prophylaxis or treatment of an RNA viral infection.

[0055] In one embodiment, such RNA viral infection is either a coronoviral infection or an influenza viral infection. Preferably, the coronoviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV and HCoV-HKU1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infection. Equally preferably, the influenza viral infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0056] In another embodiment, said RNA viral infection is selected from a group comprising infections with a virus from the families of orthomyxoviridae, picornaviridae, pneumoviridae, hepeviridae, reoviridae, togaviridae, flaviviridae, arenaviridae, filoviridae, hantaviridae and rhabdoviridae. Preferably, said viral infection is selected from a group comprising RVs, PV, HRSV, HMPV, HEV, RoV, RuV, TBEV, JEV, ZIKV, YFV, WNV, DENV, LASV, EBOV, MARV, HTNV, PUUV, DOBV, SEOV, SNV, ANDV and RABV infection. The present disclosure also relates to a method of prevention or treatment of an RNA viral infection, as defined herein, said method comprising administering a combination of an individual phenolic compound and quinine, or one of its pharmaceutically acceptable salts, to a patient in need thereof, wherein the individual phenolic compound is selected from rutin, isoquercitrin, chlorogenic acid and neochlorogenic acid, wherein, preferably, said individual phenolic compound is selected from rutin and isoquercitrin.

[0057] The present disclosure also relates to a combination of an individual phenolic compound, as defined herein, and quinine, or one of its pharmaceutically acceptable salts, for the manufacture of a medicament for the prevention or treatment of an RNA viral infection, as defined herein

[0058] DETAILED DESCRIPTION OF THE INVENTION

[0059] Elderberry is a wild-growing plant, widely distributed in Europe, Africa, Asia, and North America (cf. Dominguez et al. (2020) Food Chem 330: 127266). Elderberry fruits are small, black berries, whose color and extent resemble aronia berries and blueberries. The content of primary metabolites (sugars and organic acids) gives elderberry the typical sweet and astringent taste, while its characteristic aroma is related to the presence of different volatile and phenolic compounds. The purple color of fruit originates from anthocyanins and color pigments (cf. Ferreira et al. (2020) Food Chem 302: 125366).

[0060] The inhibitory effect of elderberry extract on the replication of SARS-CoV-2 has been disclosed in WO 2023 / 104732 and Setz et al. (2023) Nutraceuticals 3: 91-106. Cyanidin 3-glucoside, a major component in elderberry extract, was found to be effective in infections with an influenza virus, particularly in the post-infection period (Torabian et al. (2019) J Functional Foods 54: 353-360). The flavonoid kaempferol and quercetin from elderberries display an effect against the DNA virus Herpes simplex virus HSV-1 (Amoros et al. (1992) J Nat Prod 55: 1732-1740).

[0061] The term “elderberry extract”, as used herein, relates to an extract from elderberry fruits. It is preferred that this extract is an ultrafiltrate of elderberry fruits. The term “elderberry” relates to elderberry fruits. Elderberries comprise about 80 % water, 18 % carbohydrates, and less than 1 % each of protein and fat. Elderberries, preferably black elderberries, are rich in anthocyanins. An elderberry extract for use according to the invention is prepared from raw elderberries by means of ultrafiltration and is enriched with anthocyanins and / or a high molecular weight fraction comprising dietary fibers.

[0062] During ultrafiltration, the anthocyanins, the high molecular weight fraction and the dietary fibers of the elderberry raw material are retained within the extract. In one embodiment, an elderberry extract comprises about 20 wt.-% polyphenols, about 7 wt.-% dietary fibers, as well as carbohydrates and peptides. Said elderberry extract can be liquid or solid. In other embodiments, said elderberry extract has a polyphenol content of at least 2 wt.-%, of at least 3 wt.-%, or about 4.6 wt.-%. These polyphenols are mostly flavonoids and comprise at least one of rutin, quercetin, quercetin-3-O-hexoside, catechin, epicatechin, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isoquercetin, kaempferol 3-O-rutinoside, isorhamnetin 3-O-rutinoside, isorhamnetin 3- glucoside, astragalin, 5,7,3’4’-tetra-O- methylquercetin, quercetin acetylhexosides and / or quercetin hexoside pentosides (cf. Mocanu and Amariei (2022) Plants 11 : 740). In one embodiment, a liquid elderberry extract has an anthocyanin content in the range of 1 wt.-% to 10 wt.-%, preferred of 1 wt.-% to 5 wt.-%, most preferred of 2.5 wt.-% to 3 wt.-%. In one embodiment, a solid elderberry extract has an anthocyanin content in the range of 1 wt.-% to 25 wt.-%, preferred of 2.5 wt.-% to 20 wt.-%, most preferred of 5 wt.-% to 15 wt.-%. In one embodiment, a liquid extract has an anthocyanin content of 3 wt.-% to 4 wt.-%. In one embodiment, a solid extract has an anthocyanin content of 4 wt.-% to 15 wt.-%. In one embodiment, a liquid extract is obtained performing an ultrafiltration and has an anthocyanin concentration of 3.2 wt.-%. In one embodiment, a solid extract is obtained by drying, such as spray-drying, of a liquid extract obtained by ultrafiltration. In one embodiment, a solid extract is a spray-dried extract. In one embodiment, a solid extract e.g., spray-dried extract, has an anthocyanin content of 4 wt.-%, 7 wt.-%, 10 wt.-%, 14 wt.-%, or 15 wt.-%. In one embodiment, said spray dried extract is spray dried on a food additive, for example a polysaccharide such as maltodextrin. In preferred embodiments, the elderberry extract is prepared from black elder (Sambucus nigra L). In one embodiment, an elderberry extract for use according to the disclosure has an immunoregulatory potential. In one embodiment, an elderberry extract according to the disclosure comprises molecules having a molecular weight of from 400 Da to 2.000.000 Da, preferred 500 Da to 2.000.000 Da, more preferred 500 Da to 70.000 Da.

[0063] The term “ethanol extract”, as used herein, refers to an elderberry extract prepared using an extraction with ethanol. The ultrafiltrated elderberry extract according to the disclosure differs from an ethanol extract in at least the composition e.g., the ethanol extract has a higher anthocyanin concentration than the ultrafiltrated elderberry extract. For example, said ethanol extract may have an anthocyanin content of more than 25 wt.-%. Typically, ethanol extracts are obtained using chromatography columns and / or resins that specifically enrich and purify anthocyanins. Typically, when preparing an ethanol extract, ethanol is used to extract the anthocyanins from the cells and subsequently from the chromatography columns. Thus, ethanol extracts have high concentrations of anthocyanins. However, the concentrations of other components of the elderberry fruit, such as polysaccharides, vitamins, and minerals, are reduced in an ethanol extract, compared to an ultrafiltrated extract. Such reduced levels of polysaccharides, vitamins and minerals in an ethanol extract compared to an ultrafiltrated extract are, for example, due to the method of preparing the ethanol extract involving a column and solvents, such as ethanol. The ultrafiltrated extract of the invention is highly advantageous in that the method of production thereof involves water as solvent instead of ethanol as a solvent. Thereby, the preservation of comprised components is enhanced. Preparing ethanol extracts typically comprises using solvents to dissolve the active ingredients and anthocyanins from the fruit. The anthocyanins then bind to columns, are washed, and then dissolved from the column with ethanol. The ethanol is then evaporated and recovered. In contrast thereto, the ultrafiltrated extract of the invention is produced without alcoholic solvents such as ethanol. A juice is produced from the elderberry fruit raw material, and the anthocyanins are subsequently enriched by means of ultrafiltration to obtain an extract for the combination according to the disclosure.

[0064] The terms “solid extract”, “solid elderberry extract” and “elderberry extract in a solid form” are used interchangeably. Likewise, the terms “liquid extract”, “liquid elderberry extract” and “elderberry extract in a liquid form” are used interchangeably.

[0065] In one embodiment, the ultrafiltrated elderberry extract is an elderberry extract prepared by ultrafiltration of elderberry fruit juice. In one embodiment, the ultrafiltrated elderberry extract is a retentate of an ultrafiltration of elderberries. In one embodiment, the ultrafiltrated elderberry extract is a retentate of an ultrafiltration of elderberry fruit juice. In one embodiment, the ultrafiltrated elderberry extract is prepared without ultrasound treatment. In one embodiment, the ultrafiltrated elderberry extract is obtained by squeezing juice out of fresh elderberry fruits and subsequently ultrafiltrating said juice. In one embodiment, said ultrafiltrated elderberry extract is obtained without extracting pomace. In one embodiment, said ultrafiltrated elderberry extract is obtained without water extraction of pomace. In one embodiment, said ultrafiltrated elderberry extract is obtained by performing an ultrafiltration of elderberry fruit juice, particularly fresh elderberry fruit juice, and obtaining a retentate of said ultrafiltration as the ultrafiltrated elderberry extract.

[0066] The term “anthocyanin”, as used herein, relates to any anthocyanin known to a person skilled in the art, particularly water-soluble vacuolar pigments that may appear red, purple, or blue. Food plants rich in anthocyanins include i.a. , elderberry, chokeberry, black current, blueberry, raspberry, black rice and black soybean. Anthocyanins belong to the flavonoids and are generated via the phenylpropanoid pathway. Anthocyanins are found in the cell vacuole, mostly in fruits and flowers, but also occur in leaves, stems, and roots. In these parts, they are found predominantly in outer cell layers such as the epidermis and peripheral mesophyll cells. Most frequent in nature are the glycosides of cyanidin, delphinidin, malvidin, pelargonidin, peonidin, and petunidin. Sometimes anthocyanins are present in the form of a complex and / or aggregate, in which several anthocyanins are aggregated, optionally involving binding of magnesium. The terms “cyanidin”, respectively “anthocyanidin” as used herein, refer to a particular type of anthocyanin. They refer to the respective aglycon moiety in anthocyanins i.e. , without the glycoside. It is a pigment found in many red fruits including grapes, blackberry, blueberry, cherry, cranberry, elderberry, and raspberry. Cyanidin anthocyanins can be selected from cyanidin 3-glucoside, cyanidin 3-sambucoside, cyanidin 3- sambubioside 5- glucoside, cyanidin 3,5-diglucoside, and combinations thereof. Further anthocyanins are chrysanthemin, chrysanthemin monoglucuronide, antirrhinin, callistephin, tulipanin, pelargonidin-3-sambubioside, peonidin-3-glucoside, peonidin-3-sambubioside and peonidin monoglucuronide (cf. Mocanu and Amariei (2022) Plants 11 : 740). In one embodiment, the elderberry extract for use in the combination according to the disclosure is enriched with the beforementioned cyanidin anthocyanins.

[0067] In one embodiment, said ultrafiltrated extract has an anthocyanin concentration of at least 1 wt.-%, preferably at least 3 wt.-%. In one embodiment, said anthocyanin is selected from the group consisting of cyanidin 3-glucoside, cyanidin 3-sambucoside, cyanidin 3-sambubioside 5-glucoside, and combinations thereof. In one embodiment, said extract has an anthocyanin content in the range of from 2.5 wt.-% to 20 wt.-%, preferably from 3 wt.-% to 18 wt.-% e.g., 3.2 wt.-%, 4 wt.-%, 7 wt.-%, 10 wt.-%, 14 wt.-% or 15 wt.-%.

[0068] Components such as the anthocyanins and / or flavonoids comprised in the extract of the invention have an antiviral effect on the replication of SARS-CoV-2. Cyanidin 3-sambucoside has an antiviral effect on the replication of SARS-CoV-2 e.g., by blocking the enzyme neuraminidase in the viral replication process. Flavonoids have an antiviral effect on the replication of SARS-CoV-2 e.g., by blocking receptors on the virus surface. The anthocyanin concentration can be determined using any suitable method known to the person skilled in the art e.g., HPLC.

[0069] Further components in elderberry extract include phenolic acids such as caffeoylquinic acid, feruloylquinic acid and coumaroylquinic acid, organic acids such as shikimic acid, fumaric acid, citric acid, malic acid, tartric acid and valeric acid, vitamins such as ascorbic acid and retinol, lectins such as Sambucus nigra agglutinin IV (SNA-IV) and Sambucus nigra agglutinin-V (SNA-V) and sugars such as fructose and sucrose (cf. Mocanu and Amariei (2022) Plants 11 : 740).

[0070] In a final composition according to the disclosure an elderberry extract according to the disclosure may be included in a weight range of 0.01 wt.-% to 2 wt.-%, preferred 0.02 wt.-% to 1 wt.-%, more preferred 0.03 wt.-% to 0.5 wt.-%, most preferred 0.04 wt.-% to 0.1 wt.-%. The term “enriched”, as used herein, refers to an increased presence of a component in a unit of a processed composition, such as a fruit extract e.g., an ultrafiltrated elderberry extract, compared to the initial composition, such as fruit raw material. In one embodiment, the concentration of enriched components, such as anthocyanins and a high molecular weight fraction, is higher in an ultrafiltrated elderberry extract than in fruit raw material. In one embodiment, the amount of anthocyanin is enriched in an elderberry extract compared to the elderberry raw material e.g., the amount of anthocyanin is enriched in 1 kg of elderberry extract compared to 1 kg of elderberries. In one embodiment, anthocyanin is enriched in an elderberry extract at a ratio of at least 2: 1 , preferred at least 16:1 , more preferred at least 72: 1 , compared to the fruit raw material. In other words, said enrichment refers to 2 kg fruit material to 1 kg extract, 16 kg fruit material to 1 kilo extract, 72 kg fruit material to 1 kg extract, respectively. In one embodiment, the high molecular weight fraction is enriched in an elderberry extract for use in the combination according to the present disclosure, compared to the raw fruit material. In one embodiment, anthocyanin and / or the high molecular weight fraction is / are enriched in an elderberry extract for use in the combination according to the present disclosure compared to the raw fruit material e.g., at a ratio of 2:1 , 16:1 , or 72:1. The term “ratio”, as used herein, indicates a relation between a processed product, particularly an elderberry extract, and the unprocessed raw material, particularly raw elderberry fruits. The ratio indicates the amount of raw material used to prepare a certain amount of processed product. In one embodiment, when referring to a ratio of 2:1 , 16 :1 , and 72:1 , said ratios indicate that 2 kg raw fruit material, 16 kg raw fruit material, and 72 kg raw fruit material, respectively, were used to prepare 1 kg of elderberry extract. In one embodiment, when referring to an enrichment of anthocyanin at a ratio of 2:1 , 16:1 , and 72:1 , said ratios refer to the same amount of anthocyanin present in 2 kg fruit raw material, 16 kg fruit raw material, and 72 kg fruit raw material, respectively, being present in 1 kg processed product, i.e. in 1 kg of elderberry extract.

[0071] The terms “raw material”, “fruit material”, and “fruit raw material”, as used herein, refer to fruits, particularly elderberries, preferably elderberries which have not undergone extraction. In one embodiment, fruit raw material refers to unprocessed fruits. In one embodiment, fruit raw material is used to prepare an extract of the present invention by means of ultrafiltration. In one embodiment, the one or more elderberries, provided in step 1) of a method of preparing ultrafiltrated elderberry extract (see below), is fruit raw material. In one embodiment, fruit raw material refers to fresh fruits and / or frozen fruits. In one embodiment, the term “fresh fruit” may relate to a fruit such as an unprocessed fruit up to five days after harvesting, preferred up to two days after harvesting, more preferred up to 12 hours after harvesting, and / or may relate to fruits which have been frozen within 48 hours, preferred within 12 hours, more preferred within 6 hours, most preferred within 3 hours after harvesting. The term “high molecular weight fraction”, as used herein, relates to a fraction of elderberries that is obtained during ultrafiltration of said elderberries. Said high molecular weight fraction has a molecular weight in a range of from 5 kDa to 2 MDa. The composition of the high molecular weight fraction can be determined by the pore size of the membrane used for ultrafiltration. In one embodiment, said ultrafiltration membrane has a molecular weight cut-off of at least 400 Da, preferred of at least 500 Da, more preferred of at least 1000 Da. Said high molecular weight fraction usually comprises dietary fibers. Said high molecular weight fraction can comprise components such as polysaccharides, oligosaccharides, agglomerated molecules and high molecular weight proteins. The extract for use in the combination according to the disclosure comprises a high molecular weight fraction having a molecular weight of at least 5 kDa, preferably in a range of from 5 kDa to 2 MDa. In one embodiment, all components of the high molecular weight fraction have a molecular weight of at least 5 kDa, preferably in a range of from 5 kDa to 2 MDa. Said extract has a high molecular weight fraction content of at least 1 wt.-%, preferred at least 3 wt.-%, more preferred in the range of 5 wt.-% to 10 wt.-%. In one embodiment, said high molecular weight fraction comprises or consists of components that have a molecular weight that exceeds the molecular weight cut-off of at least 400 Da, preferably of at least 500 Da, more preferably of at least 1000 Da. In one embodiment, said high molecular weight fraction comprises aggregates of compounds that are smaller than the molecular weight cut-off of at least 500 Da, preferably of at least 400 Da, while other aggregates have a molecular weight that exceeds the molecular weight cut-off of at least 400 Da, preferably of at least 500 Da, more preferably of at least 1000 Da. In one embodiment, said high molecular weight fraction comprises proteins, dietary fibers, and / or polysaccharides. The high molecular weight fraction of the extract is highly advantageous in that it comprises polysaccharides that have an immunostimulatory effect. An advantage of the extract for use in the combination according to the disclosure is that it comprises a high molecular weight fraction comprising polysaccharides having an inhibitory effect on viral replication of SARS-CoV-2.

[0072] In one embodiment, the term “dietary fiber”, as used herein, refers to any dietary fiber known to the person skilled in the art, particularly a portion of fruits that cannot be completely catabolized by digestive enzymes in the human body. Dietary fibers may comprise soluble fibers and insoluble fibers. Soluble fibers are water-soluble and are fermented in the colon into gases and physiologically active by-products such as short-chain fatty acids produced by intestinal bacteria in the colon. Soluble fibers may delay gastric emptying which can result in an extended feeling of satiety. Insoluble fibers are not degraded by digestive enzymes in the upper gastrointestinal tract and may absorb water on their way through the intestines. Insoluble fibers may ease defecation. In one embodiment, dietary fibers are selected from polysaccharides, cellulose, starch, dextrins, inulin, lignin, chitin, pectins, oligosaccharides, and combinations thereof. Consuming dietary fibers has several advantages e.g., the food volume is increased without providing a high caloric content, thereby providing satiety which brings along a reduced appetite. Further, they absorb water and form a viscous gel during digestion, thereby delaying the absorption of glucose which lowers the fluctuation of blood sugar levels. Additionally, they lower cholesterol levels which may reduce the risk of cardiovascular diseases and they alleviate constipation symptoms. Moreover, an ultrafiltrated elderberry extract comprising dietary fibers displays an immunostimulatory effect. Said dietary fibers may comprise pectins, acidic polysaccharides and other polysaccharides, and combinations thereof. In one embodiment, said extract has a dietary fiber concentration of at least 1 wt.-%, preferably of at least 2 wt.-%. In one embodiment, an extract in a solid form has a dietary fiber concentration in the range of 2 wt.-% to 20 wt.-%, preferably of 5 wt.-% to 10 wt.-%. In one embodiment, an extract in a liquid form has a dietary fiber concentration in the range of 1 wt.- % to 10 wt.-%, preferably of 1 wt.-% to 5 wt.-%.

[0073] The term “liquid form”, as used herein, relates to a formulation of said extract, wherein the extract is liquid. Such a liquid extract usually has a viscosity of < 10 mPa s, preferably < 8 mPa s, as measured using a Mettler RM 180 Rheomat at room temperature at 500 RPM with a spindle diameter of 1.4 cm and a container diameter of 3.3 cm. The viscosity of a liquid extract can be increased by admixing a thickening agent. The term “solid form”, as used herein, relates to a formulation of said extract, wherein the extract is solid. Such an extract in solid form usually has an anthocyanin content of at least 1.5 wt.-%, preferably of at least 10 wt.-%. In one embodiment, such an extract in solid form is a powder, respectively a dried liquid extract. For example, such an extract in liquid form is dried via spray drying or lyophilization, preferably spray drying, to obtain an extract in solid form. The components of a solid extract and a liquid extract may be similar or identical except of the water content. In one embodiment, the drying factor of a solid extract compared to a liquid extract is factor 4 i.e. , 4 units of liquid extract are dried to obtain 1 unit of solid extract as a powder. For example, a liquid extract may have an anthocyanin content of 3.2 wt.-%. For example, a solid extract may have an anthocyanin content of 4 wt.-%, 7 wt.-%, 10 wt.-%, 14 wt.-%, or 15 wt.-%. In one embodiment, the anthocyanin concentration of an extract, for example a solid extract, is adjusted to a desired anthocyanin content by admixing a carrier such as maltodextrin or gum arabic. In one embodiment, a solid extract with an anthocyanin content of 15 wt.-% is adjusted to obtain a solid extract with an anthocyanin content of 4 wt.-%, 7 wt.%, 10 wt.-% or 14 wt.-% by adding a carrier such as maltodextrin or gum arabic. Such a solid extract has a moisture content in the range of 2 wt.-% to 8 wt.-%. In one embodiment, the solid extract comprises particles, wherein 98 % to 100 % of said particles have a particle size < 250 pm.

[0074] The term “isolated”, as used herein, in the context of an anthocyanin compound or a phenolic compound is meant to refer to a compound that is not part of an extract, but is or has been purified or is or has been synthesized de novo, and is being used as a monotherapy, with less than 1 % (w / w or w / v) of other anthocyanin compounds or phenolic compounds or other active pharmaceutical ingredients being present. In a preferred embodiment, such isolated anthocyanin or phenolic compound is being used with less than 0.1 % (w / w or w / v) of other anthocyanin compounds or phenolic compounds or other active pharmaceutical ingredients being present.

[0075] The phrase “is used on its own without additional other active pharmaceutical ingredients”, as used herein in the context of elderberry extract and / or quinine, or one of its pharmaceutically acceptable salts, is meant to refer to the use of such elderberry extract or quinine as a monotherapy. In such monotherapy, no other active pharmaceutical ingredient is being used for the treatment or prophylaxis that is intended to be achieved by said elderberry extract or quinine.

[0076] The term “individual”, as used herein, in the context of a combination of an “individual” anthocyanin compound or an “individual” phenolic compound and quinine, or one of its pharmaceutically acceptable salts, is meant to refer to an anthocyanin or phenolic compound that is not part of an extract, but is or has been purified or is or has been synthesized de novo, and is being used with less than 1% (w / w or w / v) of other anthocyanin compounds or phenolic compounds being present, but is being used in such combination together with quinine, or one of its pharmaceutically acceptable salts. In a preferred embodiment, such individual anthocyanin or phenolic compound is being used in such combination, with less than 0.1 % (w / w or w / v) of other anthocyanin compounds or phenolic compounds being present in such combination.

[0077] The chemical name of quinine is (-)-(8a,9R)-6'-methoxycinchonan-9-ol. Synonymous names and identifiers are 1-(6-methoxyquinoline-4-yl)-1-(5-vinyl-1-azabicyclo[2.2.2]oct-2-yl)methanol and 1-(6-methoxyquinoline-4-yl)-1-(5-vinyl-1 ,4-ethanopiperidine-2-yl)methanol).

[0078] In the scope of the present disclosure the term quinine shall include all pharmaceutically acceptable salt of quinine, if not expressly referenced otherwise.

[0079] Quinine is a quinoline alkaloid. Traditionally, quinine was obtained from quina bark (Cinchona pubescens, a rubiacea growing in high altitude forests in South America).

[0080] Quinine is a white, poorly water-soluble crystalline powder with a bitter taste. It was used long time in folk medicine for treating febrifugal diseases. Quinine was the first drug found to be effective in the treatment of malaria, in particular of the complicated form Malaria tropica. It complexes toxic ferriprotoporphyrin IX and thus inhibits the formation of non-toxic beta-hematin in the vacuoles of blood schizonts of Plasmodium falciparum. Nowadays it is still in use in the treatment of chloroquine-refractory malaria pathogens and if artemisinins are not indicated. In the USA it is often used against Malaria tropica. It is further used against babesiosis (Babesia infections).

[0081] The inhibitory effect of quinine on the replication of SARS-CoV-2 was disclosed in WO 2021 / 219244 and in GroBe et al. (2021) Viruses 13: 647. Anecdotal reports of an efficacy of quinine in influenza virus infections arose since the Spanish flu, and are now scientifically confirmed e.g., for Influenza A Virus (IAV) (cf. Marois et al. (2014) PLoS One 9: e110631). An inhibitory effect of quinine sulfate on the DNA virus HSV-1 was described in Baroni et al. (2007) J Dermatol Sc 3: P253-255).

[0082] Quinine has also analgesic, local anesthetic and antipyretic properties. Therefore, low doses of quinine are used in China for treating common colds.

[0083] When administered in low doses as quinine sulfate it also used to resolve cramps such as leg cramps at night.

[0084] It has also been in use for treating restless leg syndrome but was discontinued because of side effects.

[0085] Further indications are lupus erythematosus, osteoarthrosis and rheumatoid arthritis.

[0086] Quinine can be provided as pharmaceutically acceptable salts of organic and inorganic acids. Suitable examples are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, citric acid, oxalic acid, malonic acid, salicylic acid, p-aminosalicylic acid, malic acid, fumaric acid, succinic acid, ascorbic acid, maleic acid, sulfonic acid, phosphonic acid, perchloric acid, nitric acid, formic acid, propionic acid, gluconic acid, digluconic acid, lactic acid, tartaric acid, hydroxymaleic acid, pyruvic acid, phenylacetic acid, benzoic acid, p- aminobenzoic acid, p-hydroxybenzoic acid, dinitrobenzoic acid, chlorobenzoic acid, methanesulfonic acid, ethanesulfonic acid, nitric acid, hydroxyethanesulfonic acid, ethylenesulfonic acid, p-toluylsulfonic acid, naphthylsulfonic acid, sulfanilic acid, camphorsulfonic acid, alginic acid, capric acid, hippuric acid, pectinic acid, phthalic acid, quinic acid, mandelic acid, o-methyl mandelic acid, hydrogen benzenesulfonic acid, picric acid, adipic acid, cyclopentane propionic acid, D-o-toluyl tartaric acid, tartronic acid, benzenesulfonic acid, alpha-methyl benzoic acid, (o, m, p-)methyl benzoic acid, naphthylamine sulfonic acid, as well as salts from other mineral acids or carbonic acids well known to a person skilled in the art. These salts are generated by contacting the free base with a sufficient amount of the respective acid in order to build the salt in a conventional manner.

[0087] Frequently used salts of quinine include quinine sulfate, quinine dihydrochloride, quinine gluconate, quinine hydrochloride and quinine sulfate dihydrate. Preferred are quinine sulfate and quinine dihydrochloride. Further salts of quinine are revealed in US 2009 / 239900 A1. Pharmaceutically acceptable salts should be seen in terms of this application as an active agent containing a compound according to the invention in form of a salt, in particular if this salt bestows specific or ameliorated pharmacokinetic properties in comparison to the free form of the active agent or to another salt of the active agent. The pharmaceutically acceptable salt of the active agent may also bestow a pharmacokinetic characteristic to the active agent it did not have in its free form. Thus, it may even positively influence the pharmacodynamics of the active agent in respect to its therapeutic efficacy in the organism.

[0088] Adverse side effects of quinine include thrombocytopenia, thrombotic microangiopathy and methemoglobinemia. Long-term therapy with high doses of quinine sulfate may lead to nausea, headache, sweating, tinnitus, visual disturbances, fever, hypotension, hemolytic anemia, acute kidney injury, liver toxicity, blindness and disorders of the gastrointestinal tract, the dermis, the cardiovascular system (low platelet count, hemolytic-uremic syndrome / thrombotic thrombocytopenic purpura (HUS / TTP), long QT syndrome and other serious cardiac arrhythmias including torsades de pointes, blackwater fever, disseminated intravascular coagulation, leukopenia, neutropenia) and the nervous system, in rare cases asthma, hemoglobinemia (cf. Liles et al. (2016) Am J Hematol 91 : 461-466).

[0089] The combination of elderberry extract and quinine shows supraadditive effects in a dosedependent manner (see Examples 2 - 9). This allows for reducing the effective dosage I amount of quinine in the combination. The beforementioned possible adverse side effects can thus be avoided or at least mitigated. Alternatively, the effective dosage I amount of quinine can be left unchanged, but a higher efficacy can be achieved in the combination according to the disclosure.

[0090] In the food industry, quinine is used as a bitter flavoring. It can be added to fancy drinks like bitter lemon or tonic water (in the European Union up to 100 mg / kg, in Germany 85 mg / kg) or to alcoholic beverages like gin tonic and bitters (in the European Union up to 300 mg / kg, in Germany 250 mg / kg).

[0091] In a final composition according to the disclosure quinine or a water-soluble pharmaceutically acceptable salt thereof may be included in a weight range of 0.01 wt.-% to 1 wt.-%, preferred 0.02 wt.-% to 0.5 wt.-%, more preferred 0.04 wt.-% to 0.3 wt.-%, most preferred 0.05 wt.-% to 0.25 wt.-%.

[0092] To avoid ambiguities in the art, the following terms of interaction among drugs are defined. They are used in this sense throughout the disclosure. If at least two substances, respectively components are administered and at least one of these components affects the activity of the other at least one substance a so-called drug-drug interaction is given. If said interaction leads to an exaggerated or increased effect of the drug substance the effect is synergistic. The respective equation can be displayed as (A+B) > A or (A+B) > B, wherein A and B are the percentual or fractional effects (i.e., a value between 0 and 1) seen when the respective components are administered alone and (A+B) is the percentual or fractional effect seen after combinatory application, respectively. A final effect equal to the sum of the effects seen in the single component applications is an additive synergistic effect. If the final effect is greater than said expected effect the term supraadditivity is used. Smaller effects are subadditive. If, on the other hand the drug-drug interaction leads to a decreased effect of the component the effect is not synergistic at all, but antagonistic.

[0093] Hence, in the scope of this disclosure the terms “antagonism” and “antagonistic” are used for any decreased drug effect induced by drug-drug-interaction, whilst the terms “synergy” and “synergistic” are used for any increased drug effect induced by drug-drug-interaction. To describe and determine the grade of said synergy the terms “subadditive”, “additive” and “supraadditive” as well as the respective nouns are used. Hence, the term supraadditivity is used to describe an effect of two or more combined components that is greater than the expected additive effects of the single component.

[0094] The use of synergistically acting combinations can both help to increase the therapeutic efficacy and the potency, wherein the latter primarily may help to reduce off-target toxicity.

[0095] The identification of drug-drug-interactions depends on the “no-interaction” null-hypotheses, which is based on the observed drug response and not on any model of mechanism. Hence, for two different components, their grade of additivity is based on a point of reference for the readout wherein the point of reference depends on the mathematical model chosen. Various methods exist to identify such interactions and to calculate the expected additive effect of two substances and thus to determine if the actual synergistic effect observed is subadditive, additive or supraadditive. Different methods are recommended in the art. The most common methods are outlined below.

[0096] Basic methods like building simple arithmetic sums or the fractional product method provide an easy way to gain first insights if a specific combination has supraadditive potential.

[0097] The simple arithmetic sum method for additivity is based on the equation A + B = (A+B), wherein A and B again are the percentual or fractional effects seen when the respective substances are administered alone and (A+B) is the percentual or fractional effect seen after combinatory application of said substances using the same doses as in the single applications. If A + B > (A+B) supraadditivity is given. An obviously striking disadvantage of this method is that results wherein A + B > 100% cannot be analyzed for supraadditivity. Hence, this method is primarily used to interpret combinations of suboptimal doses.

[0098] The fractional product method is based on the equation 1 - (1-A) * (1-B) = (A+B). Here, A and B are the fractal effects seen when the respective substances are administered alone and (A+B) is the fractal effect seen after combinatory application of said substances using the same doses as in the single applications. If 1 - (1-A) * (1-B) > (A+B), supraadditivity is given.

[0099] More sophisticated methods include the use of a so-called isobologram, a graph constructed on a coordinate system defined by individual drug doses showing a “line of additivity” that allows to distinguish between subadditive, additive (along the line) and supraadditive effects. The “line of additivity” connects those single drug doses which display the same effect (e.g., 50% inhibition of a specific marker). All possible dose combinations along this line are expected to show the same effectiveness. Dose combinations located within the triangle built by coordinates and the line of additivity i.e. , nearer to the arbitrary point, displaying the same effect are considered as supraadditive. Dose combinations located outside the triangle are considered as subadditive. Respective figures can be mapped using specific software as e.g., CompuSyn (Chou and Martin, ComboSyn, Inc. Paramus, NJ 2007 [www.combosyn.com]).

[0100] Also, further specific indicator values as for example the Combination Index (Cl, equation of Chou and Talalay (1984) Adv Enzyme Reg 22. 27-55) and the Dose-Reduction Index (DRI, Chou equation 1984) can be calculated easily using the beforementioned specific software. Both these values allow to determine if drug substances act supraadditive synergistically when administered contemporarily. The Cl is based on the principles of mass action law and is applicable for any kind of drug combination regardless of mechanism of action, dynamic order or the quantity units used for each drug in the combination. The Cl value defines synergism as supraadditive if Cl<1 and as additive if Cl =1. If Cl > 1 the effect is either subadditive or antagonistic. Hence, a Cl of 1 also refers to the “line of additivity”. In the classic isobologram as mentioned above. In a simplified approach it could be calculated as follows: Cl = A(t) / A(x) + B(t) / B(x), wherein A(t) respectively B(t) is the dose of drug A respectively B alone that inhibits x %, whereas A(x) and B(x) stand for the portion of the respective drug in the combination that also inhibits x %. The DRI is a measure of how many folds the dose of each drug in a synergistic combination may be reduced at a given effect level compared with the doses of each drug alone. Therefore, DRI = 1 indicates additivity, whereas DRI > 1 and < 1 indicate supraadditivity and subadditivity (or antagonism), respectively. For displaying purposes, the isobologram, i.e., the equi-effective curve at various concentrations or doses of two drugs as mentioned above, is a dose-oriented figure approach based on a special case of the Cl equation. A more convenient figure approach, however, is the effect oriented so-called FaCI plot, displaying the Combination Index (Cl) against the fractal effect (Fa), preferably for a specific dose combination.

[0101] An alternative approach is offered by the web-based program SynergyFinder. To evaluate the potential synergy of combined drugs, the observed drug combination responses are usually compared with expected combination responses calculated by means of synergy scoring models (so-called reference or null models). This model assumes a stochastic process whereby two small molecules develop their effects independently.

[0102] Consequently, based on deviation of observed and expected responses one can classify the drug combination either as synergistic (i.e., combination effect is higher than expected) or antagonistic (i.e., combination effect is lower than expected). One option to visualize the results is the so-called Bliss synergy score. The Bliss independence model assumes a stochastic process in which two drugs elicit their effects independently, and the expected combination effect can be calculated based on the probability of independent events. Thus, Bliss synergy score, SBHSS, is defined as: SBHSS = EA,B, ... ,N - 100(1 - (1 - EA / 100) (1 - EB / 100)...(1 - EN / 100)), Here, terms (1 - EA,B...N / 100)) represent probabilities that drugs A,B,...,N do not inhibit the target. Thus, multiplication of these terms results in the probability that none of the drugs inhibit the target. Subtracting this probability from 1 , 1 - (1 - EA / 100)(1 - EB / 100)...(1 - EN / 100)), gives the probability that at least one of the drugs inhibits the target, which is equivalent to the Bliss effect. A Bliss synergy score < -10 means that the two tested substances act antagonistically, a score between -10 and 10 represents additive activity, while a score > 10 indicates drug synergism (lanevski et al. (2017) Bioinformatics 33: 2413-2415; Liu et al. (2018) Stat Biopharm Res 10: 112-122); lanevski et al. (2020) Nucleic Acids Res 48: W488- W493; https: / / synergyfinder.fimm.fi, as of August 16th2023).

[0103] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction or inhibition of the replication of an RNA virus in a human.

[0104] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction of the viral load of an RNA virus in a human.

[0105] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an RNA virus infection in a human, wherein said human is asymptomatic.

[0106] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an RNA virus infection in a human, wherein said human shows mild RNA virus infection- related symptoms and is not in need of hospitalization.

[0107] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of an RNA virus infection in a human, wherein said human shows severe RNA virus infection-related symptoms and is in need of hospitalization. In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of an RNA virus infection in a human, wherein said human shows severe RNA virus infection-related symptoms and is undergoing acute lung injury.

[0108] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction or inhibition of the replication of a coronavirus in a human.

[0109] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction of the viral load of a coronavirus in a human.

[0110] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of COVID-19 in a human.

[0111] The severity of a COVID-19 is usually classified according to the WHO Clinical Progression Scale (published in: Lancet Infect Dis (2020) 20: e192-e197). In the scope of the present disclosure the following classifications follow this scale.

[0112] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of a coronaviral infection in a human, wherein said human is asymptomatic.

[0113] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of a coronaviral infection in a human, wherein said human shows mild coronavirus infection- related symptoms and is not in need of hospitalization.

[0114] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of a coronaviral infection in a human, wherein said human shows severe coronavirus infection-related symptoms and is in need of hospitalization.

[0115] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of a coronaviral infection in a human, wherein said human shows severe coronavirus infection-related symptoms and is undergoing acute lung injury. In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction or inhibition of the replication of an orthomyxovirus in a human.

[0116] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction of the viral load of an orthomyxovirus in a human.

[0117] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an orthomyxovirus infection in a human, wherein said human is asymptomatic.

[0118] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an orthomyxovirus infection in a human, wherein said human shows mild orthomyxovirus infection-related symptoms and is not in need of hospitalization.

[0119] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of an orthomyxovirus infection in a human, wherein said human shows severe orthomyxovirus infection-related symptoms and is in need of hospitalization.

[0120] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of an orthomyxovirus infection in a human, wherein said human shows severe orthomyxovirus infection-related symptoms and is undergoing acute lung injury.

[0121] Unless otherwise stated, any technical or scientific term used in the present invention has the meaning that a man skilled in the relevant technical art will attribute to them.

[0122] Dietary supplements are defined as nutrients provided to a consumer that otherwise may not be consumed in a sufficient quantity. It also refers to compounds that may generally improve a person’s health condition without being intended as a therapeutic means for a disease. Therefore, different regulatory requirements exist for dietary supplements and pharmaceutical drugs in almost all countries. According to the invention, these dietary supplements shall be used exclusively for nutritional purposes in substantially healthy persons. Any possible therapeutic use in a patient in need thereof or double use shall not be covered by the term dietary supplement.

[0123] The combination of elderberry extract and quinine according to the disclosure can be combined with a variety of dietary supplement additives, as laid out in the following: Suitable vitamins are for example vitamin C (L-ascorbic acid, sodium L-ascorbate, calcium L- ascorbate, potassium L-ascorbate, L-ascorbyl 6-palmitate), vitamin A (retinol, retinyl acetate, retinyl palmitate, beta-carotene), vitamin D (cholecalciferol, ergocalciferol), vitamin E (D-alpha- tocopherol, DL-alpha-tocopherol, D-alpha-tocopheryl acetate, DL-alpha-tocopheryl acetate, D- alpha-tocopheryl succinate), vitamin K (phylloquinone), vitamin B1 (thiamin hydrochloride, thiamin mononitrate), vitamin B2 (riboflavin, sodium riboflavin 5’-phosphate), niacin (nicotinic acid, nicotinamide), pantothenic acid (calcium D-pantothenate, sodium D-pantothenate, D- panthenol), vitamin B6 (pyridoxine hydrochloride, pyridoxine 5’-phosphate), folic acid (pteroyl monoglutaminic acid), vitamin B12 (cyanocobalamin, hydroxocobalamin), biotin (D-biotin).

[0124] Suitable minerals to be included are for example calcium (calcium carbonate, calcium chloride, citric acid calcium salt, calcium gluconate, calcium glycerophosphate, calcium lactate, orthophosphoric acid calcium salt, calcium hydroxide, calcium oxide), magnesium (magnesium acetate, magnesium carbonate, magnesium chloride, citric acid magnesium salt, magnesium gluconate, magnesium glycerophosphate, ortho-phosphoric acid magnesium salt, magnesium lactate, magnesium hydroxide, magnesium oxide, magnesium sulfate), iron (iron carbonate, iron citrate, iron ammonium citrate, iron gluconate, iron fumarate, iron sodium diphosphate, iron lactate, iron sulfate, iron diphosphate, ferric saccharate, elemental iron), copper (copper carbonate, copper citrate, copper gluconate, copper sulfate, copper lysine complex), iodine (sodium iodide, sodium iodate, potassium iodide, potassium iodate), zinc (zinc acetate, zinc chloride, zinc citrate, zinc gluconate, zinc lactate, zinc oxide, zinc carbonate, zinc sulfate), manganese (manganese carbonate, manganese chloride, manganese citrate, manganese gluconate, manganese glycerophosphate, manganese sulfate), sodium (sodium bicarbonate, sodium carbonate, sodium chloride, sodium citrate, sodium gluconate, sodium lactate, sodium hydroxide, ortho-phosphoric acid sodium salt), potassium (potassium bicarbonate, potassium carbonate, potassium chloride, potassium citrate, potassium gluconate, potassium glycerophosphate, potassium lactate, potassium hydroxide, ortho-phosphoric acid potassium salt), selenium (sodium selenite, sodium hydrogen selenite, sodium selenite), chrome (chrome-(lll)-chloride, chrome-(lll)-sulfate), molybdenum (ammonium molybdate (molybdenum (VI), sodium molybdate (molybdenum (VI)), fluorine (sodium fluoride, potassium fluoride), chlorine, phosphor.

[0125] Trace elements are dietary minerals that are needed by the organism in very small amounts for growth, development and physiology, for example as co-enzymes. Some of them are virtually always present in the organism in sufficient quantities, others have to be substituted in persons in need thereof. They can be selected from the group comprising, but not limited to, chrome, cobalt, iron, iodine, copper, manganese, molybdenum, selenium, zinc, fluoride, silicon, arsenic, nickel, rubidium, tin, vanadium. They can be substituted either as a pure element or in any of the mineral forms mentioned above.

[0126] The term “prophylaxis” in the scope of the present disclosure refers to an attention towards a person that might suffer from a definable disorder in the future. In the present case prophylaxis refers to the administration of a pharmaceutically active agent in the form of a drug for medical therapy or a dietary supplement to a person that might catch an RNA viral infection in the future, in particular to a person who runs an increased risk therefor. Herein the prophylaxis shall inhibit an RNA viral infection, lower the risk of such an infection and / or mitigate the disease course in case of such an infection.

[0127] The term “treatment” in the scope of the present disclosure refers to an attention towards a person that already suffers from a definable disorder. This attention can include the administration of a pharmaceutically active agent in the form of a drug for medical therapy or a dietary supplement. Herein the treatment shall cure an RNA viral infection, prevent or mitigate symptoms related to such an RNA viral infection and / or shorten the disease course.

[0128] RNA viruses are a very broad realm of viruses. They can affect humans, other animals and plants. By unfavorable mutations such RNA viruses from animals and plants can become pathogenic in humans in the future. It can be reasonably assumed that such mutations will not affect the underlying mechanism of action of the elderberry extract and / or quinine. Therefore, the present disclosure refers to all known RNA virus families. In particular, the present disclosure refers to a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts for use in the prophylaxis or treatment of an infection with an RNA virus, wherein said infection with an RNA virus is selected from an infection with a virus from a virus family consisting of coronaviridae, chrysoviridae, megabirnaviridae, quadriviridae, totiviridae, sedoreoviridae, spinareoviridae, cystoviridae, amalgaviridae, curvulaviridae, fusariviridae, hypoviridae, partitiviridae, picobirnaviridae, hadakaviridae, birnaviridae, permutotetraviridae, botybirnaviridae, polymycoviridae, alphatetraviridae, benyviridae, hepeviridae, matonaviridae, bromoviridae, closteroviridae, endornaviridae, kitaviridae, mayoviridae, togaviridae, virgaviridae, alphaflexiviridae, betaflexiviridae, gammaflexiviridae, deltaflexiviridae, tymoviridae, flaviviridae, noaviridae, sinhalaviridae, carmotetraviridae, tombusviridae, narnaviridae, mitoviridae, Atkinsviridae, Duinviridae, Fiersviridae, solspiviridae, Blumeviridae, Steitzviridae, botourmiaviridae, hypoviridae, abyssoviridae, arteriviridae, cremagaviridae, gresnaviridae, olifoviridae, medioniviridae, mesoniviridae, hexponiviridae, mononiviridae, Nanghoshaviridae, euroniviridae, roniviridae, tobaniviridae, caliciviridae, dicistroviridae, iflaviridae, marnaviridae, picornaviridae, polycipiviridae, secoviridae, solinviviridae, alvernaviridae, barnaviridae, solemoviridae, potyviridae, astroviridae, Yadokariviridae, Hadakaviridae, sarthroviridae, qinviridae, aspiviridae, aliusviridae, chuviridae, crepuscuviridae, myriaviridae, natareviridae, artoviridae, bornaviridae, filoviridae, lispiviridae, mymonaviridae, nyamiviridae, paramyxoviridae, pneumoviridae, rhabdoviridae, sunviridae, xinmoviridae, Yueviridae, arenaviridae, cruliviridae, discoviridae, hantaviridae, leishbuviridae, mypoviridae, nairovipridae, peribunyaviridae, phasmaviridae, phenuiviridae, tospoviridade, wupedeviridae, lincruviridae, amnoonviridae, orthomyxoviridae, kolmioviridae.

[0129] Particularly preferred is a use of a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts in the prophylaxis or treatment of an infection with an RNA virus, wherein the RNA virus is highly pathogenic in humans and / or is widely spread among mankind.

[0130] Such RNA viruses are in particular:

[0131] Coronaviridae (single-stranded positive-sense RNA viruses).

[0132] Coronaviral infections that can be treated with a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof are above all infections with the highly pathogenic SARS-CoV, MERS-CoV and SARS-CoV-2. But also infections with less pathogenic coronaviridae as listed in the following can be thus treated. The terms “coronavirus” or “coronaviral” refer mainly to the sub-family of orthocoronavirinae. They are subdivided into the genera of alphacoronaviruses, betacoronaviruses, gammacoronaviruses and deltacoronaviruses. Alphacoronaviruses comprise the subgenera of colacoviruses (species: bat coronavirus CDPHE15), decaviruses (bat coronavirus HKLI10, Rhinolophus ferrumequinum alphacoronavirus HuB-2013), duvinacoviruses (human coronavirus 229E), luchacoviruses (Lucheng Rn rat coronavirus), minacoviruses (Ferret coronavirus, Mink coronavirus 1), minunacoviruses (miniopterus bat coronavirus 1 , miniopterus bat coronavirus HKLI8), myotacoviruses (Myotis ricketti alphacoronavirus Sax-2011), nylactoviruses (Nyctalus velutinus alphacoronavirus SC-2013), pedacoviruses (porcine epidemic diarrhea virus, Scotophilus bat coronavirus 512), rhinacovi ruses (Rhinolophus bat coronavirus HKLI2), setracoviruses (human coronavirus NL63, NL63-related bat coronavirus strain BtKYNM63-9b) and tegacoviruses (Alphacoronavirus 1 - type species). Betacoronaviruses comprise the sub-genera of embecoviruses (Betacoronavirus 1 (subspecies: human coronavirus OC43), China Rattus coronavirus HKLI24, human coronavirus HKLI1 , murine coronavirus-type species), hibecoviruses (Bat Hp- betacoronavirus Zhejiang 2013), merbecoviruses (Hedgehog coronavirus 1 , MERS- CoV), Pipistrellus bat coronavirus HKLI5, Tylonycteris bat coronavirus HKLI4), nobecoviruses (Rousettus bat coronavirus GCCDC1 , Rousettus bat coronavirus HKLI9 and sarbecoviruses (severe acute respiratory syndrome-related coronavirus (subspecies: SARS-CoV, SARS-CoV-2). Gammacoronaviruses comprise the subgenera of cegacoviruses (Beluga whale coronavirus SW1) and igacoviruses (Avian coronavirus-type species). Deltacoronaviruses comprise the sub-genera of andecoviruses (Wigeon coronavirus HKLI20), buldecoviruses (Bulbul coronavirus HKU11-type species, Porcine coronavirus HKLI15, Munia coronavirus HKLI13, Whiteeye coronavirus HKLI16), herdecoviruses (Night heron coronavirus HKLI19) and Moordecoviruses (Common moorhen coronavirus HKLI21).

[0133] Coronaviruses pathogenic in humans are until now SARS-CoV, SARS-CoV-2, MERS-CoV and HCoV-HKLH , HCoV-NL-63, HCoV-OC43 and HCoV-229E. The last four cause only relatively mild symptoms (cf. Andersen et al.: The Proximal Origin of SARS-CoV-2, on virologica.org, as of February 17th, 2020).

[0134] Orthomyxoviridae (single-stranded negative-sense RNA viruses). This family includes i.a., influenza viruses.

[0135] Influenza virus families pathogenic in humans and other vertebrates are Alphainfluenzavirus (species: Influenzavirus A; IVA; IAV), Betainfluenzavirus (species: Influenzavirus B; IVB; IBV), Gammainfluenzavirus (species: Influenzavirus C; IVC; ICV), Deltainfluenzavirus (species: Influenzavirus D; IVD, IDV). For pathologic infections in humans IVA and IVB are of particular concern.

[0136] IAV subtypes are categorized according to their specific serotypes hemagglutinin (H) and neuraminidase (N). The most important hemagglutinin serotypes are H1 , H2, H3, H5, H7 and H9. The most important neuraminidase serotypes are N1 , N2 and N7. The most important IAV subtypes are thus H1 N1 (e.g., Spanish influenza, Russian influenza; Swine influenza), H2N2 (e.g., Asian influenza), H3N2 (e.g., Hong Kong influenza), H5N1 (e.g., Avian flu), H7N9 (e.g., Avian influenza). In the scope of the present disclosure the term IAV shall thus encompass all these subtypes.

[0137] The most important IBV subtypes are BA / ictoria lineage, B / Yamagata lineage, B / Yamaguchi lineage, B / Yokohama lineage, B / Yunnan lineage, B / Zhuhai lineage. In the scope of the present disclosure the term IBV shall thus encompass all these subtypes.

[0138] ICV subtypes differ only in an insignificantly. ICV affects mostly humans and porcines. In children ICV causes a rather harmless flu.

[0139] IDV is rather similar to ICV and infects mainly bovines and porcines. Two subtypes are known.

[0140] Picornaviridae (non-enveloped single-stranded positive sense RNA viruses). Notable examples are enterovirus (incl. rhinoviruses (RVs) and poliovirus (PV)), aphthovirus, cardiovirus and hepatovirus.

[0141] Pneumoviridae (single-stranded negative sense RNA viruses).

[0142] Notable examples are human respiratory syncytial virus (HRSV, also RSV), bovine respiratory syncytial virus (BRSV) and human metapneumovirus (HMPV).

[0143] HRSV has mainly two subtypes A and B and can infect the upper airways and cause a cold (rhinitis, cough, acute bronchitis, otitis media).

[0144] Hepeviridae (non-enveloped single-stranded positive sense RNA viruses).

[0145] Notable examples are hepevirus and hepatitis E virus (HEV).

[0146] Reoviridae (non-enveloped double-stranded RNA viruses).

[0147] A notable example is Rotavirus (RoV; species A, B, C, D, E, F, G, H, I, J) which causes diarrhea in infants and young children.

[0148] Togaviridae (enveloped single-stranded positive sense RNA viruses).

[0149] A notable example is Rubella virus (RuV) which causes rubella.

[0150] Flaviviridae (enveloped single-stranded positive sense RNA viruses).

[0151] Notable examples are tick-borne encephalitis virus (TBEV), Japanese encephalitis virus (JEV), Zika virus (ZIKV), Yellow fever virus (YFV), West Nile virus (WNV) and Dengue virus (DENV).

[0152] - Arenaviridae (enveloped single-stranded ambisense RNA viruses)

[0153] A notable example is Lassa mammarenavirus (LASV) which causes Lassa fever.

[0154] Filoviridae (enveloped single-stranded positive sense RNA viruses)

[0155] Notable examples are Zaire ebolavirus (EBOV) which causes Ebola and Marburg Marburgvirus (MARV) which causes hemorrhagic fever.

[0156] Hantaviridae (enveloped single-stranded mostly negative sense RNA viruses).

[0157] Notable examples are Hantaan virus (HTNV), Puumala virus (PUIIV), Dobrava-Belgrade virus (DOBV), Seoul virus (SEOV), Sin nombre virus (SNV) and Andes virus (ANDV).

[0158] Rhabdoviridae (enveloped single-stranded negative sense RNA viruses).

[0159] A notable example is Rabies lyssavirus (RABV) which causes rabies.

[0160] The rationale behind an efficacy of the combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts for use in the prophylaxis or treatment of an infection with an RNA virus is the innate antiviral defense in humans which recognizes the viral RNA and leads to an activation of RNAses that degrade the viral RNA and thus suppresses the replication of the RNA virus. Compounds that support, respectively stimulate this innate antiviral defense (see above) are therefore apt to combat virtually any RNA viral infection or at least mitigate their disease course. Therefore it is reasonable to assume that the shown efficacy of the combination of the disclosure against SARS-CoV-2 and IAV can be transferred to virtually any RNA viral infection.

[0161] In a preferred embodiment, the present disclosure refers to a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts for use in the prophylaxis or treatment of an infection with an RNA virus, wherein said infection with an RNA virus is selected from the group comprising the viral families of coronaviridae, orthomyxoviridae, picornaviridae, pneumoviridae, hepeviridae, reoviridae, togaviridae, flaviviridae, arenaviridae, filoviridae, hantaviridae and rhabdoviridae.

[0162] Thus, the present application relates particularly to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of a coronaviral infection, wherein said coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infections.

[0163] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction or inhibition of the replication of a coronavirus in a human, wherein said coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infections.

[0164] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction of the viral load of a coronavirus in a human, wherein said coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, H CoV-H KU 1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infections.

[0165] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of a coronaviral infection in a human, wherein said human is asymptomatic and said coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV- HKU1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infections.

[0166] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of a coronaviral infection in a human, wherein said human shows mild coronavirus infection- related symptoms, is not in need of hospitalization and said coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1 , HCoV-NL- 63, HCoV-OC43 and HCoV-229E infections.

[0167] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of a coronaviral infection in a human, wherein said human shows severe coronavirus infection-related symptoms, is in need of hospitalization and said coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infections.

[0168] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of a coronaviral infection in a human, wherein said human shows severe coronavirus infection-related symptoms, is undergoing acute lung injury and said coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-HKU1 , HCoV-NL-63, HCoV-OC43 and HCoV-229E infections.

[0169] Thus, the present application relates in particular to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of a coronaviral infection, wherein said coronaviral infection is a SARS-CoV-2 infection.

[0170] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction or inhibition of the replication of a coronavirus in a human, wherein said coronaviral infection is a SARS-CoV- 2 infection.

[0171] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction of the viral load of a coronavirus in a human, wherein said coronaviral infection is a SARS-CoV-2 infection.

[0172] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of a coronaviral infection in a human, wherein said human is asymptomatic and said coronaviral infection is a SARS-CoV-2 infection.

[0173] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of a coronaviral infection in a human, wherein said human shows mild coronavirus-infection related symptoms, is not in need of hospitalization and said coronaviral infection is a SARS- CoV-2 infection.

[0174] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of a coronaviral infection in a human, wherein said human shows severe coronavirus-infection related symptoms, is in need of hospitalization and said coronaviral infection is a SARS-CoV-2 infection.

[0175] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of a coronaviral infection in a human, wherein said human shows severe coronavirus-infection related symptoms, is undergoing acute lung injury and said coronaviral infection is a SARS-CoV-2 infection.

[0176] The other above-mentioned animal coronaviruses have not yet made a transfer to humans (zoonosis), but this may happen in the future with an unpredictable pathology. Thus, the scope of the present application relates also to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of these animal coronaviral infections in animals and humans.

[0177] The concept for treating coronaviral infections over all species is based on the structural similarity of coronaviruses. Thus, it can be assumed that treatment and / or prevention options can be transferred from one coronavirus to another. Coronavirus particles contain four main structural proteins: Spike (S), membrane (M), envelope (E) and nucleocapsid (N), all of which are encoded within the 3’ end of the viral genome.

[0178] Coronaviruses contain a non-segmented, positive-sense RNA genome of ~30 kb. The genome contains a 5' cap structure along with a 3' poly (A) tail, allowing it to act as an mRNA for translation of the replicase polyproteins. The replicase gene encoding the nonstructural proteins (nsps) occupies two-thirds of the genome, about 20 kb, as opposed to the structural and accessory proteins, which make up only about 10 kb of the viral genome. The organization of the coronavirus genome is 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. The accessory proteins are almost exclusively nonessential for replication in tissue culture; however, some have been shown to have important roles in viral pathogenesis (cf. Zhao et al. (2012) Cell Host Microbe 11 : 607-616).

[0179] The coronavirus life cycle starts with an initial attachment of the virion to the host cell by interactions between the S protein and its receptor. The sites of receptor binding domains (RBD) within the S1 region of a coronavirus S protein vary depending on the virus. The S- protein-receptor interaction is the primary determinant for a coronavirus to infect a host species and also governs the tissue tropism of the virus. Many coronaviruses utilize peptidases as their cellular receptor. It is unclear why peptidases are used, as entry occurs even in the absence of the enzymatic domain of these proteins. Many alphacoronaviruses utilize aminopeptidase N (APN) as their receptor, many betacoronaviruses such as SARS-CoV, SARS-CoV-2 and HCoV-NL63 use angiotensin-converting enzyme II (ACE2) receptors, MHV enters through CEACAM1 , and MERS-CoV binds to dipeptidyl-peptidase 4 (DPP4) to gain entry into human cells. Following receptor binding, the virus must next gain access to the host cell cytosol. This is generally accomplished by acid-dependent proteolytic cleavage of S protein by a cathepsin, TMPRRS2 or another protease, followed by fusion of the viral and cellular membranes, and ultimately the release of the viral genome into the cytoplasm.

[0180] Coronaviruses encode either two or three proteases that cleave the replicase polyproteins. They are the papain-like proteases (PLpro), encoded within nsp3, and a serine type protease, the main protease, or Mpro, encoded by nsp5. Most coronaviruses encode two PLpros within nsp3, except the gammacoronaviruses, SARS-CoV and MERS-CoV, which only express one PLpro (Mielech et al. (2014) Virus Res doi: 10.1016).

[0181] This papain-like protease (PLpro) was found in SARS-CoV to act the same way as a deubiquitinase within the human cellular ubiquitin proteasome system (UPS) (cf. Raaben et al. (2010) J Virol 84: 7869-7879). PLpro in SARS-CoV-2 has a very high homology with SARS- CoV (96.1 %, Nguyen et al. (2020) https: / / doi:org / 10:1101 / 2020.02.05.936013).

[0182] Influenza viruses have a different structure. The ribonucleoprotein of the virus particle (virion) of an influenza virus located within the lipid envelope has an approximately helical symmetry. The ribonucleoprotein is a complex of the virus genome, structural (M1 and NP) and replication-related proteins (PA, PB1 , PB2).

[0183] In the transmission electron microscope, Influenza viruses appear as spherical or spherical ellipsoidal, occasionally filamentous, enveloped virus particles with a diameter of 80 to 120 nm (Lamb and Choppin (1983) Ann Rev Biochem 52: 467-506). Two types of spike proteins are of particular serological interest in IAV and IBV: hemagglutinin (HA) and neuraminidase (NA), against which antibodies are generated after disease and, to a lesser extent, after vaccination with influenza A and B vaccine.

[0184] The genome of almost all influenza viruses consists of eight RNA segments of negative polarity. These eight RNA molecules contain the genetic information needed for replication and assembly of viral particles and occur preferentially in single-stranded form in the virion. Segmentation of the genome is also responsible for the substantial increase in genetic variability of influenza viruses via the ability to genetically reassort (also known as antigenic shift), as an exchange of segments can occur when a cell is superinfected (with a different strain of influenza). Due to the RNA-based single-stranded genome, point mutations occur frequently (so-called antigen drift), because the RNA polymerases of RNA viruses do not have an exonuclease function to correct copying errors. Both mechanisms result in escape mutations to evade the immune response, while the functions of the virion should be preserved.

[0185] Hemagglutinin (HA) is a lectin and causes agglutination of erythrocytes and mediates attachment of the virion to a host cell during infection. The attachment of hemagglutinin to a cell occurs through an attachment of a portion of the hemagglutinin molecule to sialic acids (SA) on host cell envelope proteins that function as receptors (SA receptors). These sialic acids are clustered a2,6-linked in mammals. Each hemagglutinin variant thereby fits a specific host cell receptor according to the lock-and-key principle, with each host having only a subset of all receptors used by influenza viruses. Following proteolytic activation of the hemagglutinin taken up by a cell by cellular serine peptidases and acidification of the endosome, the hemagglutinin in its second function as a fusogenic protein causes fusion with the endosomal membrane via its fusion domain to release the ribonucleoprotein into the cytosol.

[0186] Neuraminidase (NA) is responsible for hydrolyzing N-acetylneuraminic acid at cellular receptors. This results in the release of the newly formed viruses (daughter virions from the infected cells) due to replication and thus in the spread of infection both within the same organism and to other organisms.

[0187] Matrix protein 2 (M2) consists of approximately 97 amino acids, of which 24 amino acids protrude from the membrane. In influenza A viruses, the matrix protein M2 is a proton channel for acidification of the interior of the virion after endocytosis, so that the fusion domain of hemagglutinin is triggered and fusion of the viral and endosomal membranes can occur to release the ribonucleoprotein into the cytosol. In influenza B viruses, matrix protein 2 (BM2) is not an envelope protein but a soluble protein of circa 109 amino acids.

[0188] At least 219 host proteins are required for the influenza A virus replication cycle. In humans, influenza viruses are replicated in the respiratory tract of an infected individual. Human influenza viruses prefer cells of the ciliated epithelium. In contrast, avian influenza virus replicates primarily in intestinal epithelial cells. In their host, the viruses migrate through the mucus into the epithelial cells that serve as host cells. Neuraminidase, which liquefies the mucus, ensures that they do not stick to the mucus in the process. After binding of hemagglutinin to an N-acetyl-neuraminic acid on a cell surface, invagination of the virion occurs by endocytosis. In the endosome, cellular serine proteases cut the hemagglutinin into its activated form. In addition, the pH in the endosome decreases, causing acidification of the interior of the virion via the ion channel protein M2. The acidification triggers the fusion domain of hemagglutinin, causing the viral membrane to fuse with the endosomal membrane and releasing the ribonucleoprotein into the cytosol; at the same time, M1 detaches from the ribonucleoprotein, exposing the nuclear localization signals of the NP in the ribonucleoprotein.

[0189] The viral RNA is copied in IAV and IBV by the viral polymerase complex - a heterotrimer of three proteins PA, PB1 , and PB2 using the host ribonucleotides.

[0190] Through the NS2 protein, the amplified viral RNA is shuttled from the nucleus to the cytosol, where proteins are produced after RNA submission to the ribosome. The structural proteins bind to each other, the polymerase complex of PA, PB1 , and PB2 binds to the 5' and 3' ends of the viral RNA, and the NP protein binds to the remaining viral RNA, assembling the ribonucleoprotein. The viral coat proteins HA and NA assemble at the cell surface on the lipid rafts, but M2 does not. The ribonucleoprotein binds to the inside of the lipid rafts. The mechanism of budding is not yet clear. Influenza viruses are lytic viruses. They induce programmed cell death of the host cell.

[0191] In a single infected host cell, up to about 20,000 new influenza viruses can form before it then dies and subsequently the released viruses infect further neighboring cells.

[0192] Thus, the present application relates particularly to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an influenza virus infection, wherein said influenza virus infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0193] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction or inhibition of the replication of an influenza virus in a human, wherein said influenza virus infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0194] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction of the viral load of an influenza virus in a human, wherein said influenza virus infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0195] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an influenza virus infection in a human, wherein said human is asymptomatic and said influenza virus infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0196] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an influenza virus infection in a human, wherein said human shows mild influenza virus infection-related symptoms, is not in need of hospitalization and said influenza virus infection is selected from a group consisting of IAV, I BV, ICV and IDV infection.

[0197] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of an influenza virus infection in a human, wherein said human shows severe influenza virus infection-related symptoms, is in need of hospitalization and said influenza virus infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0198] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of a coronaviral infection in a human, wherein said human shows severe influenza virus infection-related symptoms, is undergoing acute lung injury and said influenza virus infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

[0199] Thus, the present application relates in particular to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an influenza virus infection, wherein said influenza virus infection is an IAV infection.

[0200] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction or inhibition of the replication of an influenza virus infection, wherein said influenza virus infection is an IAV infection.

[0201] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the reduction of the viral load of an influenza virus infection, wherein said influenza virus infection is an IAV infection.

[0202] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an influenza virus infection in a human, wherein said human is asymptomatic, wherein said influenza virus infection is an IAV infection.

[0203] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the prophylaxis or treatment of an influenza virus infection in a human, wherein said human shows mild influenza virusinfection related symptoms, is not in need of hospitalization and said influenza virus infection is an IAV infection.

[0204] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of an influenza virus infection in a human, wherein said human shows severe influenza virus-infection related symptoms, is in need of hospitalization and said influenza virus infection is an IAV infection.

[0205] In another aspect the present application refers to a combination of elderberry extract and quinine or a pharmaceutically acceptable salt thereof for use in the treatment of an influenza virus infection in a human, wherein said human shows severe influenza virus-infection related symptoms, is undergoing acute lung injury and said influenza virus infection is an IAV infection.

[0206] Suitable aromatic and flavoring substances can be added to the combination of elderberry extract and quinine. They comprise above all essential oils that can be used for this purpose. In general, this term refers to volatile extracts from plants or parts of plants with the respective characteristic smell. They can be extracted from plants or parts of plants by steam distillation.

[0207] Suitable examples are: Essential oils, respectively aromatic substances from achillea, sage, cedar, clove, chamomile, anise, aniseed, star anise, thyme, tea tree, peppermint, mint oil, menthol, cineol, borneol, zingerol, eucalyptus, mango, figs, lavender oil, chamomile blossoms, pine needle, cypress, orange, rose, rosewood, plum, currant, cherry, birch leaves, cinnamon, lime, grapefruit, tangerine, juniper, valerian, lemon, lemon balm, lemon grass, palmarosa, cranberry, pomegranate, rosemary, ginger, pineapple, guava, echinacea, ivy leave extract, blueberry, kaki, melon, alpha- or beta-pinene, alpha-pinene oxide, alpha-campholenic aldehyde, alpha-citronellol, alpha-isoamyl-cinnamic, alpha-cinnamic terpinene, alphaterpineol, alpha-terpinene, aldehyde C16, alpha-phellandrene, amyl cinnamic aldehyde, amyl salicylate, anisic aldehyde, basil, anethole, bay, benzyl acetate, benzyl alcohol, bergamot, bitter orange peel, black pepper, calamus, camphor, cananga oil, cardamom, carnation, carvacrol, carveol, cassia, castor, cedarwood, cinnamaldehyde, cinnamic alcohol, cis-pinane, citral, citronella, citronellal, citronellol dextro, citronellol, citronellyl acetate; citronellyl nitrile, citrus unshiu, clary sage, clove bud, coriander, corn, cotton seed, d-dihydrocarvone, decyl aldehyde, diethyl phthalate, dihydroanethole, dihydrocarveol, dihydrolinalool, di hydromyrcene, 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 globulus, eugenol, evening primrose, fenchol, fennel, ferniol, fish, florazon, galaxolide, geraniol, geranium, geranyl acetate, geranyl nitrile, guaiacol, guaiacwood, gurjun balsam, heliotropin, herbanate, hiba, hydroxycitronellal, i-carvone, i-methyl acetate, ionone, isobutyl quinoleine, isobornyl acetate, isobornyl methylether, isoeugenol, isolongifolene, jasmine, lavender, limonene, linallol oxide, linallol, linalool, linalyl acetate, linseed, litsea cubeba, I- methyl acetate, longifolene, mandarin, mentha, menthane hydroperoxide, menthol crystals, menthol laevo, menthone laevo, methyl anthranilate, methyl cedryl ketone, methyl chavicol, methyl hexyl ether, methyl ionone, methyl salicylate, mineral, mint, musk ambrette, musk ketone, musk xylol, myrcene, nerol, neryl acetate, nonyl aldehyde, nutmeg, orris root, paracymene, parahydroxy phenyl butanone crystals, patchouli, p-cymene, pennyroyal oil, pepper, perillaldehyde, petitgrain, phenyl ethyl alcohol, phenyl ethyl propionate, phenyl ethyl- 2methylbutyrate, pimento berry, pimento leaf, pinane hydroperoxide, pinanol, pine ester, pine, pinene, piperonal, piperonyl acetate, piperonyl alcohol, plinol, plinyl acetate, pseudo ionone, rhodinol, rhodinyl acetate, rosalin, ryu, sandalwood, sandenol, sassafras, sesame, soybean, spearmint, spice, spike lavender, spirantol, starflower, tea seed, terpenoid, terpineol, terpinolene, terpinyl acetate, tert-butylcyclohexyl acetate, tetrahydrolinalool, tetrahydrolinalyl acetate, tetrahydromyrcenol, thulasi, thymol, tomato, trans-2-hexenol, trans-anethole, turmeric, turpentine, vanillin, vetiver, vitalizair, white cedar, white grapefruit, Wintergreen etc. or mixtures thereof, as well as mixtures of menthol, peppermint and star anise oil or menthol and cherry flavor.

[0208] Preferred are natural aromatic and / or flavoring substances. In the scope of the present disclosure the term “natural aromatic and / or flavoring substances” refers to those aromatic and flavoring substances that are of natural i.e., herbal or animal origin.

[0209] In a final composition according to the disclosure aromatic and flavoring substances may be included in a weight range of 0.01 wt.-% to 2 wt.-%, preferred 0.03 wt.-% to 1 wt.-%, more preferred 0.05 wt.-% to 0.5 wt.-%, most preferred 0.07 wt.-% to 0.3 wt.-%.

[0210] For some embodiments it may be advantageous to add a sweetener to the combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts. Quinine is known to have a bitter taste which some people do not appreciate.

[0211] Suitable sweeteners can be selected from the group comprising mannitol, glycerol, acesulfame potassium, aspartame, cyclamate, isomalt, isomaltitol, saccharin and its sodium, potassium and calcium salts, sucralose, glucose, fructose, alitame, thaumatin, glycyrrhizin, neohesperidine dihydrochalcone, steviol glycosides, neotame, aspartame-acesulfame salt, maltitol, maltitol syrup, lactitol, xylitol, erythritol.

[0212] A preferred sweetener is invert sugar syrup. Invert sugar syrup consists of monomers of glucose and fructose and is produced by hydrolytic saccharification of sucrose. It is 1.3 x sweeter than sucrose and crystallizes less easily.

[0213] In a final composition according to the disclosure a sweetener may be included in a weight range of 1 wt.-% to 25 wt.-%, preferred 3 wt.-% to 20 wt.-%, more preferred 5 wt.-% to 18 wt- %, most preferred 10 wt.-% to 15 wt.-%. For some embodiments it may be advantageous for improving the taste to regulate the pH of the combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts.

[0214] Suitable pH-regulators for liquid dosage forms are e.g., sodium hydroxide, hydrochloric acid, buffer substances such as sodium dihydrogen phosphate or disodium hydrogenphosphate.

[0215] Suitable acidity regulators can be selected from the group comprising 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, sodium potassium tartrate, ortho-phosphoric acid, lecithin citrate, magnesium citrate, ammonium malate, sodium malate, sodium hydrogen malate, calcium malate, calcium hydrogen malate, adipic acid, sodium adipate, potassium adipate, ammonium adipate, succinic acid, sodium fumarate, potassium fumarate, calcium fumarate, ammonium fumarate, 1 ,4- heptonolactone, triammonium citrate, ammonium ferric 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.

[0216] A preferred acidity regulator is crystalline anhydric citric acid.

[0217] In a final composition according to the disclosure a pH regulator and / or acidity regulator may be included in a weight range of 0.01 wt.-% to 2 wt.-%, preferred 0.05 wt.-% to 1.5 wt.-%, more preferred 0.08 wt.-% to 1 wt.-%, most preferred 0.1 wt.-% to 0.5 wt.-%.

[0218] When the combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts is provided in an aqueous solution to the final consumer it may be advantageous to include a preservative in the aqueous solution for keeping a bacterial contamination low.

[0219] Suitable preservatives may be selected from the group comprising, but not limited to, sorbic acid, potassium sorbate, sodium sorbate, calcium sorbate, methyl paraben, ethyl paraben, methyl ethyl paraben, propyl paraben, benzoic acid, sodium benzoate, potassium benzoate, calcium benzoate, heptyl p-hydroxybenzoate, sodium methyl para-hydroxybenzoate, sodium ethyl para-hydroxybenzoate, sodium propyl para-hydroxybenzoate, benzyl alcohol, benzalkonium chloride, phenylethyl alcohols, cresols, cetylpyridinium chloride, chlorbutanol, thiomersal (sodium 2-(ethylmercurithio) benzoic acid), sulfur dioxide, sodium sulfite, sodium bisulfite, sodium metabisulfite, potassium metabisulfite, potassium sulfite, calcium sulfite, calcium hydrogen sulfite, potassium hydrogen sulfite, biphenyl, orthophenyl phenol, sodium orthophenyl phenol, thiabendazole, nisin, natamycin, formic acid, sodium formate, calcium formate, hexamine, formaldehyde, dimethyl dicarbonate, 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-(ll)- sulfate, chlorine, chlorine dioxide and other suitable substances or compositions known to the person skilled in the art.

[0220] The addition of a sufficient amount of antioxidants may be preferable for liquid products containing a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts is provided. Suitable examples for antioxidants include sodium metabisulfite, alpha-tocopherol, ascorbic acid, maleic acid, sodium ascorbate, ascorbyl palmitate, butylated hydroxyanisol, butylated hydroxytoluene, fumaric acid or propyl gallate. Preferred is the use of sodium metabisulfite, alpha-tocopherol and ascorbyl palmitate.

[0221] For adjusting the consistency, respectively the viscosity of a final product containing the combination of an elderberry extract and quinine it may be advantageous to add a thickening agent. Suitable examples for thickening agents are polyvinyl pyrrolidone, methyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, dextrins, polydextrose, modified starch, alkaline modified starch, bleached starch, oxidized starch, enzyme-treated starch, monostarch phosphate, distarch phosphate esterified with sodium trimetaphosphate or phosphorus oxychloride, phosphate 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, hydroxy propyl distarch glycerin, hydroxypropyl distarch phosphate, hydroxypropyl distarch glycerol, starch sodium octenyl succinate, acetylated oxidized starch, hydroxyethyl cellulose.

[0222] In another aspect the present disclosure refers thus to a final product that includes at least one of preservative, antioxidant or thickening agent in the total range of 0 wt.-% to 5 wt.-%, preferred 0.1 wt.-% to 3 wt.-%, more preferred 0.3 wt.-% to 2 wt.-%, most preferred 0.5 wt.-% to 1.5 wt.-%.

[0223] The present application discloses therefore a Composition containing the following ingredients: elderberry in a weight range of 0.01 wt.-% to 2 wt.-%; quinine or a water-soluble pharmaceutically acceptable salt thereof in a weight range of 0.01 wt.-% to 1 wt.-%; a pH regulator and / or acidity regulator in a weight range of 0.01 wt.-% to 2 wt.-%; a sweetener in a weight range of 1 wt.-% to 25 wt.-%; one or more aromatic and flavoring substances in a weight range of 0.01 wt.-% to 2 wt.-%; and water in a weight range of 1 .4 wt.-% to 68 wt.-%, wherein the relative weight ranges of all included components sum up to 100%.

[0224] The present disclosure refers further to a composition as described before, further comprising at least one of preservatives, antioxidants, or thickening agents in a total range of 0 wt.-% to 5 wt.-% of the composition, wherein the relative weight ranges of all included components sum up to 100%.

[0225] In another aspect the present disclosure relates to a composition for use in the prophylaxis or treatment of an RNA viral infection, wherein said composition contains a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts, a carrier and at least one pharmaceutically acceptable excipient.

[0226] In general, one or more pharmaceutically acceptable carriers are added to the at least one pharmaceutically active agent. Eligible are all carriers known in the art and combinations thereof. In solid dosage forms they can be for example plant and animal fats, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silica, talcum, zinc oxide. For liquid dosage forms and emulsions suitable carriers are for example solvents, solubilizing agents, emulsifiers such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol, cotton seed oil, peanut oil, olive oil, castor oil, sesame oil, glycerol fatty acid esters, polyethylglycols, fatty acid esters of sorbitan. Suspensions may use carriers known in the art such as diluents (e.g., water, ethanol or propylene glycol), ethoxylated isostearyl alcohols, polyoxyethylene and polyoxyethylene sorbitan esters, microcrystalline cellulose, bentonites, agar agar, tragacanth.

[0227] The term “excipient” is used in the present disclosure to describe any component of a pharmaceutical composition apart of the at least one pharmaceutically active principle. The selection of suitable excipients depends on a variety of factors, such as the dosage form, the dosage, the desired solubility and the stability of the composition.

[0228] The term “pharmaceutically acceptable excipient(s)” refers to natural or synthetic compounds that are added to a pharmaceutical formulation alongside the pharmaceutical active agent. They may help to bulk up the formulation, to enhance the desired pharmacokinetic properties or the stability of the formulation, as well as being beneficial in the manufacturing process. Advantageous classes of excipients according to the invention include binding agents, colorants, buffers, preservatives, antioxidants, coatings, sweeteners, thickening agents, pH- regulators, acidity regulators, acidifiers, solvents, isotonizing agents, disintegrants, glidants, lubricants, emulsifiers, solubilizing agents, stabilizers, diluents, anti-caking agents (antiadherents), sorbents, foaming agents, anti-foaming agents, opacifiers, fatliquors, consistency enhancers, hydrotropes, aromatic and flavoring substances. These compounds are known to an expert in the field.

[0229] The present disclosure refers further to a pharmaceutical composition as described before for use in medicine.

[0230] A juice containing the combination according to the disclosure of elderberry extract and quinine or one of its pharmaceutically acceptable salts can be obtained with the following method, comprising the steps of: a) Providing elderberries from Sambucus nigra L. and / or Sambucus canadiensis, b) Optionally, heating and / or thawing of said elderberries, c) Optionally, enzymatically treating said elderberries, d) Extracting anthocyanins from said elderberries by water extraction for affording an elderberry water extract, e) Optionally, pasteurizing and / or cooling said elderberry water extract, f) Ultrafiltrating said elderberry water extract by using an ultrafiltration membrane with a molecular weight cut-off of at least 400 Da, thereby enriching said anthocyanins in said elderberry water extract, g) Optionally, pasteurizing and / or cooling said ultrafiltrated elderberry water extract, h) Adding quinine or one of its pharmaceutically acceptable salts to said ultrafiltrated elderberry water extract, i) Optionally, adding at least one of aromatic and flavoring substances, sweeteners, pH- regulators, acidity regulators, preservatives, antioxidants, or thickening agents, and j) Obtaining a juice comprising ultrafiltrated elderberry water extract and quinine or one of its pharmaceutically acceptable salts.

[0231] In a preferred embodiment, said enzymatically treatment of said elderberries in step c) of the beforementioned method is carried out by a pectolytic enzyme.

[0232] In a preferred embodiment, said extraction of anthocyanins from said elderberries by water extraction in step d) of the beforementioned method is carried out by adding water and mechanically disrupting said elderberries by juicing, mashing, grinding, shredding, chopping, pressing, blending and / or pulping said elderberries.

[0233] In a preferred embodiment, said ultrafiltration membrane in step f) of the beforementioned method has a molecular weight cut-off of at least 500 Da, in a more preferred embodiment of at least 1 kDa. Pectolytic enzymes are a group of enzymes that break down pectin, a polysaccharide found in plant cell walls, through hydrolysis, transelimination and deesterification reactions. They can be selected from a group comprising pectolyase, pectozyme, polygalacturonase, pectinases PB1 , PB2, PB3, pr combinations thereof (cf. Sakai et al. (1993) Adv Appl Microbiol 39: 213- 294).

[0234] EXAMPLES

[0235] Liquid European black elderberry extract (provided by Iprona AG / SpA, Lana, Italy; charge EC3.2) and quinine sulfate (Sigma-Aldrich, Darmstadt, Germany) were prepared freshly before the experiments.

[0236] Example 1 : Analysis of the used European black elderberry extract charge EC3.2

[0237] The main components of the European black elderberry extract charge EC3.2were determined. To this aim, the anthocyanin and polyphenol content of the extract was analyzed by means of HPLC (High Performance Liquid Chromatography).

[0238] It showed that cyanidin-3-glucoside, cyanidin-3-sambubioside, cyanidin-3-sambubioside-5- glucoside, chlorogenic acid and rutin were by far the most frequent components.

[0239] Example 2: Combinations of European black elderberry extract charge EC3.2 and quinine sulfate inhibit the replication of SARS-CoV-2 in infected Calu-3 cells in a supraadditive manner in qRT-PCR qRT-PCR (quantitative real-time polymerase chain reaction) experiments were conducted. This method is a very sensitive method to determine quantitatively the number of SARS-CoV- 2 RNA copies in infective virions released from cells in the supernatant. The method was performed according to Corman et al. (2020) Euro Surveill 25: 2000045. Calu-3 cells (cf. Park et al. (2021) Biomol Ther 29: 273-281) were received from Prof. Jan Munch, Institute of Molecular Virology, University of Ulm, Germany. They were maintained in Minimal Essential Medium (MEM) containing 20% (v / v) inactivated FCS (fetal calf serum), 1 mM L-glutamine, 100 U / mL penicillin, and 100 pg / mL streptomycin and 1 mM sodium pyruvate.

[0240] Treatment scheme:

[0241] Day 0:

[0242] Seeding of Calu-3 cells in 24-well plates (1*105cells / well; use 500 pl medium per well)

[0243] Day 1 : Infection and treatment

[0244] Post-Treatment

[0245] - Infection of confluent monolayers of Calu-3 cells with SARS-CoV-2 for 1 hour in FCS- free DM EM with a 100-fold dilution of the field isolate SARS-COV-2PF (isolated from a 61 years old patient six days after the presumed date of infection and two days after start of mild COVID-19 symptoms)

[0246] - Discard medium

[0247] - Add new medium with substances (500 pl) for 3 days

[0248] Day 3: Preparation of virion containing supernatants for qRT-PCR

[0249] Take 200 pl supernatants in 1 .5 ml Eppendorf reaction tubes

[0250] Centrifuge at 6000 rpm for 5 minutes

[0251] Transfer the supernatant in a new 1.5 ml Eppendorf reaction tube

[0252] Heat for 10 min at 95°C qRT-PCR analysis with 5 pl

[0253] Per experiment, each treatment was run four times. At harvest, respective supernatants will be pooled, corresponding to n=1.

[0254] Samples containing the released virions were quantified by real-time PCR AgPath-ID One- Step RT-PCR Kit from Ambion (Cat: 4387424), allowing reverse transcription, cDNA synthesis and PCR amplification in a single step. The gene sequence amplified by qRT-PCR was from RdRp (viral RNA-dependent RNA polymerase), a prominent gene in RNA viruses.

[0255] The sequences of the used qRT-PCR primers were:

[0256] RdRp_fwd: 5 -GTG-ARA-TGG-TCA-TGT-GTG-GCG-G-3' ; “R” denoting a purine nucleotide, i.e. A or G

[0257] RdRp_rev: 5 -CAR-ATG-TTA-AAS-ACA-CTA-TTA-GCA-TA-C-3' “R” denoting a purine nucleotide, i.e. A or G

[0258] Probe: 5 --CAG-GTG-GAA- / ZEN / CCT-CAT-CAG-GAG-ATG-C -3' (Label: FAM / IBFQ Iowa Black FQ). “ZEN” denoting an internal quencher As positive control a specific target for E and RdRp gene of SARS-CoV-2 was used and made by Integrated DNA Technologies:

[0259] Control: 5'-TAA-TAC-GAC-TCA-CTA-TAG-GGT-ATT-GAG-TGA-AAT-GGT-CAT-GTG-TGG- CGG-TTC-ACT-ATA-TGT-TAA-ACC-AGG-TGG-AAC-CTC-ATC-AGG-AGA-TGC-CAC-AAC- TGC-TTA-TGC-TAA-TAG-TGT-TTT-TAA-CAT-TTG-GAA-GAG-ACA-GGT-ACG-TTA-ATA- GTT-AAT-AGC-GTA-CTT-CTT-TTT-CTT-GCT-TTC-GTG-GTA-TTC-TTG-CTA-GTT-ACA- CTA-GCC-ATC-CTT-ACT-GCG-CTT-CGA-TTG-TGT-GCG-TAC-TGC-TGC-AAT-ATT-GTT- 3'

[0260] Samples were analyzed by 7500 software v2.3 (Applied Bioscience). Plots were generated with Microsoft® Excel and Prism GraphPad-Software.

[0261] The following concentrations, respectively combinations were tested:

[0262] European black elderberry extract charge EC3.2 alone was dissolved in aqua bidest. in the following ratios (w / w): a) 1 :3200 b) 1 :1600 c) 1 :800

[0263] Quinine sulfate alone: a) 0.01 pM b) 0.1 pM c) 1 pM

[0264] All these concentrations were combined with each other. The results for the combinations of said three dissolutions of European black elderberry extract charge EC3.2with 0.01 pM quinine sulfate are depicted in Fig. 1A, for the combinations of said three dissolutions of European black elderberry extract charge EC3.2 with 0.1 pM quinine sulfate are depicted in Fig. 1 B, and for the combinations of said three dissolutions of European black elderberry extract charge EC3.2 with 1 pM quinine sulfate are depicted in Fig. 1C, all in relation to untreated cells which were set to 100%.

[0265] With this very sensitive method a dose-dependent reduction in the amount of SARS-Cov-2 RNA copies could be shown for European black elderberry extract charge EC3.2 and quinine sulfate as well as for all combinations thereof. Further, all combination showed a supraadditive effect, in comparison to the corresponding concentrations of European black elderberry extract charge EC3.2 and quinine sulfate as single substances.

[0266] These experiments show that not only the single components European black elderberry extract charge EC3.2and quinine sulfate inhibit dose-dependently the replication of SARS-CoV- 2 in Calu-3 cells but that also all tested combinations are able to do so, in a dose-dependent manner and with a supraadditive efficacy, n = 3; data are represented as mean + SEM. Example 3: Combinations of European black elderberry extract charge EC3.2 and quinine sulfate in effective concentrations are not cytotoxic in Calu-3 cell cultures

[0267] For addressing the question whether combinations of European black elderberry extract charge EC3.2 and quinine sulfate show cytotoxic effects in the abovementioned systems noninfected Calu-3 cells were treated in parallel with increasing ratios of combinations of European black elderberry extract charge EC3.2 (dissolution 1 :3200, 1 :1600, 1 :800) and quinine sulfate (0.01 pM, 0.1 pM, 1 pM). Toxicity is assessed with a WST (water-soluble tetrazolium salt 1) assay. Herein viable cells with an intact mitochondrial succinate-tetrazolium dehydrogenase system effect an enzymatic conversion of the feebly red tetrazolium salt WST-1 (4-[3-(4- iodophenyl)-2-(4-nitrophenyl)-2 / 7-5-tetrazolio]-1 ,3 benzene disulfonate) into dark red formazan. This color change can be measured photometrically in a spectrophotometer. Thus, the WST assay is a very sensitive method for measuring the toxicity of substances on the cell metabolism. The value for untreated cells was set to 100%.

[0268] It can be shown that combinations of European black elderberry extract charge EC3.2 and quinine sulfate do not display any significant toxic effect in antiviral ly effective concentrations in Calu-3 cells during an observation period of 3 days.

[0269] In Fig. 2 the percentage of viable cells is depicted in comparison to untreated cells. The value for untreated cells was set to 100%. 1 pM staurosporine (an indolocarbazole compound from Streptomyces staurosporeus, an apoptosis inducer) was used as positive control, n = 3; data are represented as mean + SEM.

[0270] Thus, it can be stated that the effects on SARS-CoV-2 replication of combinations of European black elderberry extract charge EC3.2 and quinine sulfate are not due to unspecific cytotoxic effects.

[0271] Example 4: The CompuSyn software for additive effects confirms the supraadditive effect on SARS-CoV-2 replication for all combinations of European black elderberry extract charge EC3.2 and quinine sulfate

[0272] As described before, the CompuSyn software model for additive effects (www.combosyn.com) is a computer modulation for evaluating additive effects of two substances in a biological system. By mathematical transformations the results of the respective pharmaceutical combinations of Example 2 were related to the results for both single substances.

[0273] Fa / Cl plots for all tested pharmaceutical combinations (non-constant ratio) The abscissa displays the fractal effect (Fa) i.e., a value between 0 and 1 for the relative inhibition of the respective pharmaceutical combination of European black elderberry extract charge EC3.2 and quinine sulfate on the replication of SARS-CoV-2 in Calu-3 cells, wherein 1 means a 100% inhibition and 0 means no inhibition. The ordinate displays the Combination Index (Cl) as calculated via CompuSyn Software, wherein the value 1 shows additivity, values smaller than 1 show supraadditivity and values above 1 show subadditivity, no effect or antagonism. The closer the values are to 0 the more prominent is the supraadditive effect.

[0274] This software generated the plot of Fig. 3. It shows that all combinations display a marked supraadditive effect. This computer evaluation corroborates the results of the evaluation of Example 2.

[0275] Example 5: The SynergyFinder software for additive effects confirms the supraadditive effect on SARS-CoV-2 replication for all combinations of European black elderberry extract charge EC3.2 and quinine sulfate

[0276] As described before, the SynergyFinder model for additive effects is a computer modulation for evaluating additive effects of two substances in a biological system. By mathematical transformations the results of the respective pharmaceutical combinations of Example 1 were related to the results for both single substances.

[0277] From the data of Example 2 a Bliss synergy score of 27.86 is calculated. This means that the combination of European black elderberry extract charge EC3.2 and quinine sulfate show a highly synergistic effect (> 10) on the replication of SARS-CoV-2.

[0278] Example 6: Combinations of European black elderberry extract charge EC3.2 and quinine sulfate inhibit the replication of influenza virus A PR-8 (Puerto Rico / 8 / 34 H1 N1) (MOI 0.01) in infected MDCKII (Madin-Darby canine kidney II) cells in a supraadditive manner in qRT-PCR

[0279] The Influenza A virus isolate (IAV) A / Puerto Rico / 8 / 34 (PR8) was received from Prof. Dr. Matthias Tenbusch (Institute of Clinical and Molecular Virology, Erlangen, Germany). Viral titers of each variant were determined by a plaque assay. For the generation of new virus stock, virus containing cell culture supernatant was harvested 48 hours post infection (hpi) and passed through a 0.45 pm pore-size filter. All virus stocks were stored at -80 °C until further usage. qRT-PCR (quantitative real-time polymerase chain reaction) experiments were conducted.

[0280] This method is a very sensitive method to determine quantitatively the number of IAV RNA copies in infective virions released from cells in the supernatant. The method was performed as described in Example 2.

[0281] MDCKII cells were maintained in Dulbecco’s Minimal Essential Medium (DMEM) containing 10% (v / v) inactivated FCS (fetal calf serum), 1 mM L-glutamine, 100 U / rnL penicillin and 100 pg / mL streptomycin.

[0282] For infection experiments, MDCKII cells were inoculated with IAV PR8 (multiplicity of infection (MOI): 0.01) for 30 minutes, washed and further treated with the indicated interventions. 48 hpi, virus-containing cell culture supernatants were incubated for 10 min at 95°C and finally used for qRT-PCR analysis. For titer determination of IAV PR8 virus stocks, MDCKII cells were infected with serial dilutions of the virus stock for 3 h. Afterwards cells were overlayed with 1% purified agar (Oxoid, Wesel, Germany) and incubated at 37°C for 4 days. Endpoint of virus infection was analyzed by counting plaque forming units (PFU) visualized following incubation of the cells with MTT (3-(4,5-dimethylthiazol-2-yl) for 3 hours. Viral titer was calculated by the method of Reed and Muench (1938) Am J Hyg 27 493-497).

[0283] Per experiment, each treatment was run four times. At harvest, respective supernatants will be pooled, corresponding to n=1.

[0284] Samples containing the released virions were quantified by real-time PCR GoTaq® probe qPCR one step Kit (Promega, Madison, Wisconsin, USA), allowing reverse transcription, cDNA synthesis and PCR amplification in a single step. The gene sequence amplified by qRT-PCR was from Integrated DNA Technologies, Coralville, Iowa, USA.

[0285] Sequences from the IAV matrix gene used as qRT-PCR primers were: 5’lnf-A-M: 5’-AGA TGA GTC TTC TAA CCG AGG TCG-3’

[0286] 3'lnf-A-M: 5’-TGC AAA AAC ATC TTC AAG TCT CTG-3’

[0287] 3'lnf-A-SW-M: 5’-TGC AAA GAC ATC TTC CAG TCT CTG-3’.

[0288] Samples were analyzed by 7500 software v2.3 (Applied Bioscience, Waltham, Massachusetts, USA). Plots were generated with Microsoft® Excel and Prism GraphPad-Software.

[0289] The following concentrations, respectively combinations were tested:

[0290] European black elderberry extract charge EC3.2 alone was dissolved in aqua bidest. in the following ratios (w / w): a) 1 :800 b) 1 :400 c) 1 :200

[0291] Quinine sulfate alone: a) 0.1 pM b) 1 pM The selected concentrations were combined with each other. The results for the combinations of said three dissolutions of European black elderberry extract charge EC3.2 with 0.1 pM quinine sulfate are depicted in Fig. 4A, for the combinations of said three dissolutions of European black elderberry extract charge EC3.2 with 1 pM quinine sulfate are depicted in Fig. 4B, all in relation to untreated cells which were set to 100%.

[0292] With this very sensitive method a dose-dependent reduction in the amount of IAV RNA copies could be shown for European black elderberry extract charge EC3.2 and quinine sulfate as well as for all combinations thereof. Further, all combination showed a supraadditive effect, in comparison to the corresponding concentrations of European black elderberry extract charge EC3.2 and quinine sulfate as single substances.

[0293] These experiments show that not only the single components European black elderberry extract charge EC3.2and quinine sulfate inhibit dose-dependently the replication of SARS-CoV- 2 in MDCKII cells but that also all tested combinations are able to do so, in a dose-dependent manner and with a supraadditive efficacy, n = 3; data represented as mean + SEM.

[0294] Example 7: Combinations of European black elderberry extract charge EC3.2 and quinine sulfate in effective concentrations are not cytotoxic in MDCKII cell cultures

[0295] For addressing the question whether combinations of European black elderberry extract charge EC3.2 and quinine sulfate show cytotoxic effects in the abovementioned systems noninfected MDCKII cells were treated in parallel with increasing ratios of combinations of European black elderberry extract charge EC3.2 (dissolution in water 1 :800, 1 :400 and 1 :200 wt.-%) and quinine sulfate (0.1 pM, 1 pM). Toxicity is assessed with a WST (water-soluble tetrazolium salt 1) assay. Herein viable cells with an intact mitochondrial succinate-tetrazolium dehydrogenase system effect an enzymatic conversion of the feebly red tetrazolium salt WST- 1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2 / 7-5-tetrazolio]-1 ,3 benzene disulfonate) into dark red formazan. This color change can be measured photometrically in a spectrophotometer. Thus, the WST assay is a very sensitive method for measuring the toxicity of substances on the cell metabolism. The value for untreated cells was set to 100%.

[0296] It can be shown that combinations of European black elderberry extract charge EC3.2 and quinine sulfate do not display any significant toxic effect in antiviral ly effective concentrations in MDCKII cells during an observation period of 2 days.

[0297] In Fig. 5 the percentage of viable cells is depicted in comparison to untreated cells. The value for untreated cells was set to 100%. 1 pM staurosporine (an indolocarbazole compound from Streptomyces staurosporeus, an apoptosis inducer) was used as positive control, n = 3; data are represented as mean + SEM.

[0298] Thus, it can be stated that the effects on IAV replication of combinations of European black elderberry extract charge EC3.2 and quinine sulfate are not due to unspecific cytotoxic effects.

[0299] Example 8: The CompuSyn software for additive effects confirms the supraadditive effect on IAV replication for all combinations of European black elderberry extract charge EC3.2 and quinine sulfate

[0300] The data of Example 6 were evaluated with the CompuSyn software as described in Example 4. This software generated the plot of Fig. 6. It shows that all combinations display a marked supraadditive effect. This computer evaluation corroborates the results of the evaluation of Example 6.

[0301] Example 9: The SynergyFinder software for additive effects confirms the supraadditive effect on IAV replication for all combinations of European black elderberry extract charge EC3.2 and quinine sulfate

[0302] The data of Example 6 were evaluated with the SynergyFinder software as described in Example 5. From the data a Bliss synergy score of 14.736 is calculated. This means that the combination of European black elderberry extract charge EC3.2 and quinine sulfate shows a highly synergistic effect (> 10) on the replication of IAV.

[0303] Example 10: Production of a final product containing a combination of elderberry extract and quinine

[0304] The following components are mixed in any order under stirring at room temperature and atmospheric pressure: The result is a liquid dietary supplement containing a combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts.

[0305] Eldercraft® 14% is an elderberry extract according to the disclosure which is provided by Iprona AG / SpA, Lana, Italy.

[0306] Example 11 : Experiments using anthocyanin compounds, on their own, on cells being SARS- CoV-2 infected

[0307] Anthocyanin compounds (cyanidine-3-O-sambubioside chloride, cyanidine-3 glucoside and cyanidine-3-O-sambubioside-5-O-glucoside) were used on their own to treat SARS-CoV-2- infected Calu-3 cells.

[0308] Treatment was as described in Example 2, with each of cyanidine-3-O-sambubioside chloride, cyanidine-3 glucoside and cyanidine-3-O-sambubioside-5-O-glucoside being used at concentrations of 0.01 pM, 0.1 pM, 1 pM and 10 pM, respectively. As a control, EC 3.2 (1 :100 diluted in cell culture medium) was used.

[0309] The tested compounds, when used on their own, exhibit good antiviral activity against SARS- CoV-2, with the IC50-values of cyanidine-3-O-sambubioside chloride and cyanidine-3 glucoside being ~ 2 log levels lower than for cyanidine-3-O-sambubioside-5-O-glucoside. Results are shown in Figure 7.

[0310] Example 12: Experiments using phenolic compounds, on their own, on cells being SARS- CoV-2 infected

[0311] Phenolic compounds (rutin, isoquercitrin, chlorogenic acid and neochlorogenic acid) were used on their own to treat SARS-CoV-2-infected Calu-3 cells.

[0312] Treatment was as described in Example 2, with each of rutin, isoquercitrin, chlorogenic acid and neochlorogenic acid being used at concentrations of 0.01 pM, 0.1 pM, 1 pM and 10 pM, respectively. As a control, EC 3.2 (1 :100 diluted in cell culture medium) was used.

[0313] The tested compounds, when used on their own, exhibit good antiviral activity against SARS- CoV-2, especially rutin and isoquercitrin. Results are shown in Figure 8A for rutin and isoquercitrin, and in Figure 8B for chlorogenic acid and neochlorogenic acid. A summary of IC50-values for both Examples 11 and 12 can be seen in the following table, with results for some of the compounds not being shown in any figure, because these compounds did not show any pronounced antiviral activity against SARS-CoV-2:

[0314] Example 13: Anthocyanin compounds and phenolic compounds, when used on their own, in effective concentrations are not cytotoxic in Calu-3 cell cultures

[0315] For addressing the question whether the compounds identified in Examples 11 and 12 as being antivirally active show cytotoxic effects in the abovementioned systems, non-infected Calu-3 cells were treated in parallel with increasing concentrations of these compounds using the methodology of Example 3. 10pM Staurosporine was used as a positive control.

[0316] The tested compounds are those shown in Figures 7 and 8. Results are shown in Figure 9. It can be seen that none of the compounds with antiviral activity, when used on their own, had an effect on the cell viability of non-infected Calu-3 cells. Moreover, in view of the compounds’ respective IC50-values there is a therapeutic window of at least 3 log stages that is available for the highly effective compounds. Example 14: Elderberry extract monotherapy and quinine monotherapy inhibits the replication of influenza virus A PR-8 (Puerto Rico / 8 / 34 H1 N1) (MOI 0.01) in infected MDCKII (Madin-Darby canine kidney II) cells

[0317] MDCKII cells were infected with the I AV isolate A / Puerto Rico / 8 / 34 (PR8) at a MOI of 0.01 . 30 minutes after infection and removal of input virus, cells were treated with indicated dilution steps of EC 3.2 (A) or concentrations of quinine-sulfate (B). DMSO as a solvent control, was tested at the same concentration as used in the highest concentration of quinine (10 pM). Cell culture supernatants were harvested at 2 days post infection (dpi). The virions were purified and analyzed by qRT-PCR. Bars show mean values of three independent experiments ± standard deviation. Statistical analysis was performed using a multiple comparison Kruskal- Wallis test (Anova) followed by Dunn's post hoc test (*p <0.00312; **** p <0.0001 and ns = not significant versus the untreated control). Results are shown in Figure 10. Treatment with EC 3.2 or quinine led to a dose dependent reduction of virus replication. Thereby, the production of progeny virions was blocked by 80 % at a dilution of 1 :200 EC 3.2 and by 90 % with a concentration 10 pM quinine. DMSO, a solvent control for quinine, was added at the same amount as used at the highest concentration of quinine (10 pM) had no influence on the replication of IAV (Fig. 10 B). The IC50 were - 1 :400 for EC 3.2 and -250 nM for quinine.

[0318] It becomes clear that both elderberry extract and quinine, when used on their own, have a pronounced inhibitory effect on the replication of IAV.

[0319] Example 15: Elderberry extract monotherapy and quinine monotherapy in effective concentrations are not cytotoxic in MDCKII cell cultures

[0320] For addressing the question whether Elderberry extract monotherapy and quinine monotherapy show cytotoxic effects in the abovementioned systems non-infected MDCKII cells were treated in parallel with increasing concentrations of these compounds using the methodology of Example 7. Results are shown in Figure 11. Following treatment with different dilutions of elderberry extract or concentrations of quinine (indicated at the x-axis) for two days, the influence on cell viability was measured by water-soluble tetrazolium salt (WST)-1 assay, as described in Example 7. Bars represent means of 3 independent experiments ±SD. Staurosporine (StS, 1 pM) was used as a positive control. Thereby, it was demonstrated that treatment with elderberry extract at dilutions up to 1 :100 and quinine up to 100 pM and thus in a concentration range where the replication of IAV PR8 was blocked, had no impact on cell viability (Fig. 11). The TD50 values for EC 3.2 were -1 :30 in and -500 pM for quinine. There is a 3 log therapeutic window for quinine and a 2 log therapeutic window for elderberry extract. Moreover, reference is made to the figures which show the following:

[0321] FIGURES

[0322] Fig. 1

[0323] Combination experiments with a SARS-CoV-2 infection (Example 2)

[0324] Q: quinine sulfate

[0325] E: elderberry extract charge EC3.2

[0326] %: percentage of copies of qRT-PCR-amplified RdRp (viral RNA-dependent RNA polymerase after, in comparison to untreated cells

[0327] A: bars for untreated cells (set to 100 %)

[0328] 0.01 pM quinine sulfate elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest. elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest. elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0329] 0.01 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest.

[0330] 0.01 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest.

[0331] 0.01 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0332] B: bars for untreated cells (set to 100 %)

[0333] 0.1 pM quinine sulfate elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest. elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest. elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0334] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest.

[0335] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest.

[0336] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest. C: bars for untreated cells (set to 100 %)

[0337] 1 pM quinine sulfate elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest. elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest. elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0338] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest.

[0339] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest.

[0340] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0341] Fig. 2 Cytotoxic experiments in Calu-3 cells (Example 3)

[0342] Q: quinine sulfate

[0343] E: elderberry extract charge EC3.2

[0344] S: staurosporine

[0345] %: percentage of surviving cells in the WST assay after 3 days bars (n = 3; mean ± SEM) for untreated cells (set to 100 %)

[0346] 0.01 pM quinine sulfate

[0347] 0.1 pM quinine sulfate

[0348] 1 pM quinine sulfate elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest. elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest. elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0349] 0.01 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest.

[0350] 0.01 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest.

[0351] 0.01 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest. 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest. 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest. 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest. 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :3200 in aq. bidest.

[0352] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :1600 in aq. bidest.

[0353] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0354] 1 pM staurosporine Fig. 3

[0355] Fa I Cl diagram of the results of Example 1 , generated with CompuSyn software

[0356] Fa: Fractal effect

[0357] Cl: Combination Index

[0358] Fig. 4

[0359] Combination experiments with an IAV infection (Example 6)

[0360] Q: quinine sulfate

[0361] E: elderberry extract charge EC3.2

[0362] %: percentage of copies of qRT-PCR-amplified RdRp (viral RNA-dependent RNA polymerase after, in comparison to untreated cells)

[0363] A: bars for untreated cells (set to 100 %)

[0364] 0.1 pM quinine sulfate elderberry extract charge EC3.2 dissolution 1:800 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:400 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:200 in aq. bidest.

[0365] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0366] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :400 in aq. bidest.

[0367] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :200 in aq. bidest.

[0368] B: bars for untreated cells (set to 100 %)

[0369] 1 pM quinine sulfate elderberry extract charge EC3.2 dissolution 1:800 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:400 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:200 in aq. bidest.

[0370] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0371] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :400 in aq. bidest.

[0372] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :200 in aq. bidest.

[0373] Fig. 5 cytotoxic experiments MDCKII cells (Example 7) Q: quinine sulfate

[0374] E: elderberry extract charge EC3.2

[0375] S: staurosporine

[0376] %: percentage of surviving cells in the WST assay after 2 days bars (n = 3; mean ± SEM) for control (untreated cells, set to 100 %)

[0377] 0.1 pM quinine sulfate

[0378] 1 pM quinine sulfate elderberry extract charge EC3.2 dissolution 1:800 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:400 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:200 in aq. bidest.

[0379] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0380] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :400 in aq. bidest.

[0381] 0.1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :200 in aq. bidest.

[0382] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :800 in aq. bidest.

[0383] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :400 in aq. bidest.

[0384] 1 pM quinine sulfate + elderberry extract charge EC3.2 dissolution 1 :200 in aq. bidest.

[0385] 1 pM staurosporine

[0386] Fig.6

[0387] Fa I Cl diagram of the results of Example 1 , generated with CompuSyn software

[0388] Fa: Fractal effect

[0389] Cl: Combination Index

[0390] Fig. 7

[0391] Experiments using single anthocyanin compounds on their own with a SARS-CoV-2 infection (Example 11)

[0392] E: elderberry extract charge EC3.2

[0393] %: percentage of copies of qRT-PCR-amplified RdRp (viral RNA-dependent RNA polymerase after, in comparison to untreated cells bars for untreated cells (set to 100 %)

[0394] 0.01 pM anthocyanin compound 0.1 pM anthocyanin compound

[0395] 1 pM anthocyanin compound

[0396] 10 pM anthocyanin compound elderberry extract charge EC3.2 dissolution 1:100 in aq. bidest.

[0397] The anthocyanin compound used was either cyanidine-3-O-sambubioside-5-O- glucoside, cyanidine-3-O-sambubioside chloride, or cyanidine-3-glucoside, as indicated in the figure.

[0398] Fig. 8 A

[0399] Experiments using single phenolic compounds on their own with a SARS-CoV-2 infection (Example 12)

[0400] E: elderberry extract charge EC3.2

[0401] %: percentage of copies of qRT-PCR-amplified RdRp (viral RNA-dependent RNA polymerase after, in comparison to untreated cells

[0402] A: bars for untreated cells (set to 100 %)

[0403] 0.01 pM phenolic compound

[0404] 0.1 pM phenolic compound

[0405] 1 pM phenolic compound

[0406] 10 pM phenolic compound elderberry extract charge EC3.2 dissolution 1:100 in aq. bidest.

[0407] The phenolic compound used was either rutin or isoquercitrin, as indicated in the figure.

[0408] Fig. 8 B

[0409] Experiments using single phenolic compounds on their own with a SARS-CoV-2 infection (Example 12)

[0410] E: elderberry extract charge EC3.2

[0411] %: percentage of copies of qRT-PCR-amplified RdRp (viral RNA-dependent RNA polymerase after, in comparison to untreated cells

[0412] B: bars for untreated cells (set to 100 %) 0.01 pM phenolic compound

[0413] 0.1 pM phenolic compound

[0414] 1 pM phenolic compound

[0415] 10 pM phenolic compound elderberry extract charge EC3.2 dissolution 1 :100 in aq. bidest.

[0416] The phenolic compound used was either chlorogenic acid or neochlorogenic acid, as indicated in the figure.

[0417] Fig. 9 Cell viability experiments in Calu-3 cells (Example 13)

[0418] S: staurosporine

[0419] %: percentage of surviving cells in the WST assay after 3 days bars (n = 3; mean ± SEM) for untreated cells (set to 100 %)

[0420] 0.1 pM anthocyanin compound or phenolic compound

[0421] 1 pM anthocyanin compound or phenolic compound

[0422] 10 pM anthocyanin compound or phenolic compound

[0423] 100 pM anthocyanin compound or phenolic compound

[0424] 10 pM staurosporine

[0425] The anthocyanin compound used was either cyanidine-3-O-sambubioside-5-O- glucoside, cyanidine-3-O-sambubioside chloride, or cyanidine-3-glucoside, as indicated in the figure.

[0426] The phenolic compound used was either rutin, isoquercitrin, chlorogenic acid or neochlorogenic acid, as indicated in the figure.

[0427] Fig. 10 Experiments using elderberry extract monotherapy or quinine monotherapy with an IAV infection (Example 14)

[0428] Q: quinine sulfate

[0429] E: elderberry extract charge EC3.2

[0430] %: percentage of copies of qRT-PCR-amplified RdRp (viral RNA-dependent RNA polymerase after, in comparison to untreated cells) A: bars for untreated cells (set to 100 %) elderberry extract charge EC3.2 dissolution 1:800 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:400 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:200 in aq. bidest.

[0431] B: bars for untreated cells (set to 100 %)

[0432] 0.01 pM quinine sulfate

[0433] 0.1 pM quinine sulfate

[0434] 1 pM quinine sulfate

[0435] 10 pM quinine sulfate

[0436] DM SO as solvent control

[0437] Fig. 11 cytotoxic experiments MDCKII cells (Example 15)

[0438] Q: quinine sulfate

[0439] E: elderberry extract charge EC3.2

[0440] StS: staurosporine

[0441] %: percentage of surviving cells in the WST assay after 2 days

[0442] A: bars (n = 3; mean ± SEM) for control (untreated cells, set to 100 %) elderberry extract charge EC3.2 dissolution 1:800 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:400 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:200 in aq. bidest. elderberry extract charge EC3.2 dissolution 1:50 in aq. Bidest StS = 1pM staurosporine.

[0443] B: bars (n = 3; mean ± SEM) for untreated cells (set to 100 %)

[0444] 0.01 pM quinine sulfate

[0445] 0.1 pM quinine sulfate

[0446] 1 pM quinine sulfate

[0447] 10 pM quinine sulfate

[0448] 100 pM quinine sulfate

[0449] StS = 1pM staurosporine.

Claims

Claims1. Combination of elderberry extract and quinine or one of its pharmaceutically acceptable salts.

2. Combination according to claim 1 for use as a dietary supplement.

3. Combination according to claim 1 for use in the prophylaxis or treatment of an RNA viral infection.

4. Combination for use according to claim 3, wherein the RNA viral infection is a coronaviral infection.

5. Combination for use according to claim 4, wherein the coronaviral infection is selected from a group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV and HCoV-HKLH, HCoV-NL-63, HCoV-OC43 and HCoV-229E infection.

6. Combination for use according to claim 3, wherein the RNA viral infection is an influenza viral infection.

7. Combination for use according to claim 6, wherein the influenza viral infection is selected from a group consisting of IAV, IBV, ICV and IDV infection.

8. Combination for use according to claim 3, wherein the RNA viral infection is selected from a group comprising infections with a virus from the families of orthomyxoviridae, picornaviridae, pneumoviridae, hepeviridae, reoviridae, togaviridae, flaviviridae, arenaviridae, filoviridae, hantaviridae and rhabdoviridae.

9. Combination for use according to claim 8, wherein said viral infection is selected from a group comprising RVs, PV, HRSV, HMPV, HEV, RoV, RuV, TBEV, JEV, ZIKV, YFV, WNV, DENV, LASV, EBOV, MARV, HTNV, PUUV, DOBV, SEOV, SNV, ANDV and RABV infection.

10. Combination for use according to any one of claims 1 to 9, further comprising at least one of aromatic and flavoring substances, sweeteners, pH-regulators, acidity regulators, preservatives, antioxidants, or thickening agents.

11. Composition containing the following ingredients: elderberry in a weight range of 0.01 wt.-% to 2 wt.-%; quinine or a water-soluble pharmaceutically acceptable salt thereof in a weight range of 0.01 wt.-% to 1 wt.-%; a pH regulator and / or acidity regulator in a weight range of 0.01 wt.-% to 2 wt.-%; a sweetener in a weight range of 1 wt.-% to 25 wt.-%; one or more aromatic and flavoring substances in a weight range of 0.01 wt.-% to 2 wt.-%; and water in a weight range of 1.4 wt.-% to 68 wt.-%, wherein the relative weight ranges of all included components sum up to 100%.

12. Composition according to claim 11 , further comprising at least one of preservatives, antioxidants, or thickening agents in a total range of 0 wt.-% to 5 wt.-% of the composition, wherein the relative weight ranges of all included components sum up to 100%.

13. Composition according to claim 11 containing the following ingredients: elderberry extract (0.06 wt.-%) a water-soluble pharmaceutically acceptable quinine salt (0.0104 wt.-%) anhydric citric acid (0.25 wt.-%) invert sugar syrup (13 wt.-%) flavoring agents (0.1 wt.-%), filled up to 100 wt.-% with water.

14. Pharmaceutical composition for use in the prophylaxis or treatment of an RNA viral infection, wherein said composition contains a combination as defined in claim 1, a carrier and at least one pharmaceutically acceptable excipient.

15. Pharmaceutical composition according to claim 14 for use in medicine.

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