EP4125918A1

EP4125918A1 - Virustatic agent

Info

Publication number: EP4125918A1
Application number: EP21713967.4A
Authority: EP
Inventors: Johannes Reinmüller
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-03-27
Filing date: 2021-03-25
Publication date: 2023-02-08
Also published as: WO2021191342A1; US20230122776A1

Abstract

The invention relates to the use of hydroxymethyl group donors, particularly hyaluronic acid containing hydroxymethyl groups, for treating and preventing infection with enveloped viruses such as coronaviruses, paramyxoviruses such as RS viruses, or orthomyxoviruses such as influenza viruses, and/or for treating or preventing inflammatory disease of the respiratory tract such as COPD, ARDS or cystic fibrosis, particularly inflammatory disease of the respiratory tract associated with viral infection.

Description

Antiviral agent

description

The present invention relates to the use of hydroxymethyl group donors, in particular hyaluronic acid containing hydroxymethyl groups, for the treatment and prevention of infection with enveloped viruses such as coronaviruses, paramyxoviruses, for example RS viruses, or orthomyxoviruses, for example influenza viruses, and / or for treatment or prevention an inflammatory disease of the respiratory tract, for example COPD, ARDS or cystic fibrosis, in particular also an inflammatory disease of the respiratory tract associated with a viral infection.

A large number of diseases of the respiratory tract in humans and in higher vertebrates are caused by enveloped viruses, such as influenza viruses (family Orthomyxoviridae), coronaviruses (family Coronaviridae), or RS viruses (respiratory syncytial viruses, family Paramyxoviridae). Half of all so-called colds or diseases of the human respiratory tract are caused by rhinoviruses (family Picornaviridae). In contrast to the viruses mentioned above, they do not have a lipid envelope and consist of RNA with a surrounding capsid. (They can be detected in the laboratory by PCR testing and thus differentiated from other viral pathogens.)

These viruses are mainly spread through droplet infection (aerosols) and through contact with contaminated surfaces (smear infections). Infection of the infected organism requires the pathogen to penetrate the specific host cells (entry) or to dock on their surface and the activity of viral and / or cellular enzymes for smuggling. The prevention of the viral invasion into the host cells can thus prevent the onset of the disease.

The course of an infection of the respiratory tract depends on individual factors and ranges from symptom-free to life-threatening conditions and lethal death. So far, no specific drug-antiviral treatments are known. In practice, treatment is symptomatic. In the event of survival, the various viral diseases leave behind highly specific immunities. Therefore, protection from such diseases can currently only be achieved by vaccination - provided that experience with the pathogen and a vaccine is available. A vaccine usually takes several years to develop.

However, the aforementioned virus families are subject to constant genomic change, so that in the course of successive epidemics, new pathogen variants arise time and again, for which no vaccine is available in advance. It is therefore of particular interest to develop a treatment method that either less specifically or unspecifically prevents the uptake of viruses or the multiplication of ingested viruses in the organism and thus prevents the outbreak or a severe course of the disease.

This can be achieved by modifying proteins of the virus capsid and / or the virus envelope and / or the receptor of the target cell, which are interactively essential for the virus to dock and penetrate the target cell. This prevents the virus from penetrating the target cell on the lung surface and preventing it from multiplying.

The virus capsid of the families mentioned regularly consists of proteins and proteoglycans. The cellular receptors are often proteins and proteoglycans with enzyme activity (peptidases, metallocarboxypeptidases, proteases) or sialic acids. Blocking the enzymatically active domains by transferring hydroxymethyl groups can thus prevent the viral pathogen from penetrating the cytoplasm of the target cell and thus preventing it from multiplying and spreading.

Secondary amines (arginine residues or histidine residues of serine proteases or catalytic triads) as well as amino and / or hydroxyl groups of glycosaminoglycans (including sialic acids) or proteoglycans come into consideration as acceptors of the hydroxymethyl groups. Amino groups often form the active centers of peptidases / proteases (catalytic triads), which act as cellular receptors or as virus ligands in most known coronavirus infections. They therefore play a key role in infection. The inhibition of aminopeptidases on the cell surface with alpha-ketoamides and the blocking of virus-cell contact at the ACE2 receptor by an analog are known. So far, no approved active ingredient (virustatic) for the treatment of humans has been derived from this.

In WO2012 / 168462 it is described that glycosaminoglycan derivatives which have been modified with hydroxymethyl groups have an excellent anti-infective effect against different types of pathogens. This anti-infective effect is due to the presence of hydroxymethyl groups on the glycosaminoglycan.

Surprisingly, it was found that hydroxymethyl donors against enveloped viruses, in particular coronaviruses, such as the novel coronavirus SARS-CoV 2, paramyxoviruses such as RS viruses, or orthomyxoviruses such as influenza viruses, or non-enveloped viruses, in particular picornaviruses such as rhinoviruses, have a virustatic and / or virucidal effect.

The inhibition of functional proteins of the cell surface and / or of the virus capsid including the "spikes" is achieved by transferring hydroxymethyl groups, which are brought to the location of the current or threatened virus infection - here surfaces of the upper and deep respiratory tract - by means of a suitable carrier molecule will. It is currently assumed that the mechanism of action of a hydroxymethyl group donor is based on the transfer of hydroxymethyl groups to functional groups, i.e. acceptors such as serine proteases on the cell surface of the target cell and / or on acceptors of the virus capsid, e.g. arginine clusters of "spikes" or glycosaminoglycans, is based.

This means that there is a wide range of treatments for various viral pathogens and their mutants, provided they use similar infection routes. In this way, the penetration of these pathogens, e.g. coronaviruses, into the target cell is prevented and / or slowed down. Also of benefit is an anti-inflammatory effect of hydroxymethyl donors, which is found when administered to the surfaces of the respiratory tract. This anti-inflammatory effect is based on the transfer according to the invention of hydroxymethyl groups to the serine proteases of the inflammation cascade, which is thereby inhibited. An object of the invention is thus the use of a

Hydroxymethyl group donor as an active ingredient for treating and preventing infection with viruses, in particular enveloped viruses such as coronaviruses, paramyxoviruses, e.g. RS viruses, or orthomyxoviruses, e.g. influenza viruses, or of non-enveloped viruses such as picornaviruses, e.g. rhinoviruses.

Another object of the invention is the use of a

Hydroxymethyl group donors as an active ingredient for the treatment and prevention of inflammatory diseases of the respiratory tract, for example COPD, ARDS or cystic fibrosis, in particular also of inflammatory diseases of the respiratory tract associated with viral infections. The viral infections include respiratory infections with enveloped viruses as described above, but also infections with non-enveloped viruses such as rhinoviruses.

The principle on which the present invention is based is consistent with the findings of Hoffmann et al. (Cell 181 (April 2020), 1-10). According to this, SARS-CoV-2 viruses penetrate into a target cell (epithelium) in the event of an infection, whereby the spike protein of the virus is cleaved by the cell-based TMPRSS2 protease. This protease is a serine protease. Serine proteases are significantly involved in many physiological processes, e.g. blood coagulation and inflammation, and have identical active centers (serine-histidine-aspartic acid, or so-called catalytic triad). The blockade of TMPRSS2 is currently considered to be the preferred therapeutic approach.

In the event of a SARS CoV-2 infection, the transfer of hydroxymethyl groups can inhibit the serine protease TMPRSS2 of the target cell and / or change the arginine-rich domain of the viral S protein (“spike”), possibly with the consequence a general blockade for cleavage by other proteases.

S. Buonvino, and S. Melino in "New Consensus pattem in Spike CoV-2: potential implications in coagulation process and cell-cell fusion." Cell death discovery 6.1 (November 2020): 1-5, describe the occurrence of a consensus motif in the spike protein of corona viruses. It is characterized by 13 amino acids such as cysteine and serine, which have a high affinity for hydroxymethyl groups due to their -SH or -OH end groups. The therapeutic principle pursued here uses a hydroxymethyl donor, eg taurolidine and / or modified hyaluronic acid, and is based on known, approved pharmaceutical preparations - similar to the serine protease inhibitor Camostat Mesilate. The substances mentioned act as serine protease inhibitors and thus virustatic in the sense of entry inhibitors.

In a preferred embodiment, the invention relates to the use of a hydroxymethyl group donor for the prophylaxis or treatment of an infection with coronaviruses, e.g. SARS-CoV-2.

It is assumed that the hydroxymethyl donors according to the invention, in particular taurolidine, can also inactivate the spike protein of coronaviruses by binding to the consensus motif and thus inhibit the entry of the viruses into target cells, e.g. into cells of the pulmonary alveoli.

From J. Reinmüller, “Influence of physiological and pathological coagulation by taurolidine and implications for use”, Zentralblatt Chirurgie, 124 (1999) Suppl. 4: 13-18, it is also known that taurolidine inhibits the activity of blood coagulation factor X. Factor X is a protein that is involved in the blood coagulation cascade and also contains the consensus motif.

It has recently become known that the production of the spike protein in human cells during the outbreak of Covid-19 disease and when vaccinated with a genetic vaccine (e.g. an mRNA or vector vaccine) can trigger increased blood clotting, which is caused by the formation of thrombi in the blood vessels is marked. The consensus motif can also be found on PEAR1 (Platelet endothelial aggregation receptor 1), which causes platelets to adhere to the vessel wall.

As a side effect of a corona vaccination with the AstraZeneca vaccine, thromboses in the brain are currently described, which are supposed to be caused by the aggregation of platelets in cerebral veins. Since this complication occurs shortly after vaccination, it is plausible that the consensus motif occurs in excess in the organism, eg in the blood, through the formation of spike protein as a result of the AstraZeneca vaccination, and directly or indirectly supports the PEAR1 effect. This can be an adhesion of platelets to the wall of veins and thus the development of a thrombosis.

Hydroxymethyl donors, in particular taurolidine, can have an anti-coagulatory effect by inhibiting the spike protein itself and / or the body's own proteins, e.g. factor X and / or PEAR1, and thereby prevent and / or at least reduce the formation of thrombi.

This results in a new antiviral preparation with a hydroxymethyl group donor as an active ingredient. Preferred active ingredients are biopolymers containing hydroxymethyl groups, e.g. glycosaminoglycans, especially hyaluronic acid, proteoglycans or carbohydrates, or low molecular weight substances such as taurolidine. The composition is preferably in the form of an aerosol or rinsing solution for use against the spread of the new coronavirus in the respiratory tract and for prophylaxis.

The antiviral composition can expediently be administered, e.g. inhaled as an aerosol, as soon as the first symptoms or in the case of a positive test of a patient based on a throat swab - i.e. before the onset of the disease or before the occurrence of serious symptoms - in order to spread the infection into deeper sections to inhibit the respiratory tract.

To produce the active ingredient according to the invention, a carrier substance, for example a glycosaminoglycan such as hyaluronic acid, can be reacted with a hydroxymethyl donor, for example formaldehyde, with hydroxymethyl groups being transferred to the glycosaminoglycan, for example hyaluronic acid. Suitable examples of hydroxymethyl donors are aqueous and / or alcoholic formalin solutions and / or paraformaldehyde dispersions. This means that the carrier substance, for example hyaluronic acid itself, becomes a hydroxymethyl donor in vivo. Other compounds capable of transferring hydroxymethyl groups, such as taurolidine, N-methylol-caprolactam, N-methylol-pyrrolidone, N-methylolated ureas or thioureas, methylolated, are also possible as hydroxymethyl donors, e.g. with regard to the conversion of hyaluronic acid Dicyandiamide, methylolated melamine, etc. A preferred hydroxymethyl donor is taurolidine, which is itself known to be virustatic and / or virucidal. Taurolidine comes as a dilute solution in a concentration e.g. from 0.01 to 1.0% (w / v), in particular from 0.01 to 0.5% (w / v) or from 0.1 to 0.2% (w / v) as an alternative to wetting surfaces of the respiratory tract. Due to the rapid disintegration of the molecule in biological tissue, the half-life is shorter than that of hyaluronic acid containing hydroxymethyl groups. Because of the potential for the transfer of hydroxymethyl groups to functional structures of the virus and the cell wall, taurolidine acts according to the invention and is therefore included in the invention for the treatment of viral diseases of the respiratory tract by the virus families mentioned.

Instead of hyaluronic acid, other carrier molecules can be used that contain amino and / or hydroxyl groups, such as cellulose and hydroxyethyl starch, or other glycosaminoglycans, proteoglycans, long-chain carbohydrates such as glycogen, starch, polyalcohols such as ethylene glycol, polyethylene glycols or glycerol and derivatives thereof, including chitin (e.g. chitosamine), mono-, di- or oligosaccharides, amino alcohols and suitable peptides of different composition and chain length.

A quantitative detection of removable hydroxymethyl groups in the carrier molecule can be carried out by known methods, e.g. by the chromotropic acid reaction or by the Schiff's sample.

In a preferred embodiment of the invention, the hydroxymethyl group donor is a modified glycosaminoglycan, i.e. a polysaccharide built up linearly from repeating modified disaccharides, in particular hyaluronic acid. The individual disaccharide units consist of a uronic acid which is 1 to 3 glycosidically linked to an amino sugar such as N-acetyl-glucosamine. The disaccharide units of the chains themselves are linked 1-> 4 glycosidically.

Preferred examples of glycosaminoglycans used according to the invention are glycosaminoglycans substituted on amino and / or hydroxyl groups with hydroxymethyl groups. In addition to hyaluronic acid, further examples of suitable glycosaminoglycans are heparin, chondroitin sulfate, dermatan sulfate and keratan sulfate. According to the invention, chitosamine and poly-N-acetyl-glucosamine are also understood as glycosaminoglycans. The glycosaminoglycan is preferably a hyaluronic acid. Glycosaminoglycans are usually obtained from protein-containing biological tissues. For example, the isolation of hyaluronic acid from cockscomb or from streptococci is known. Natural heparins are extracted from pig small intestinal mucosa, among other things. Chondroitin sulfate is largely obtained from the cartilage tissue of cattle, pigs and sharks. In addition, glycosaminoglycans can be produced from genetically modified host organisms such as bacterial cells.

In medicine, hyaluronic acid and its derivatives are used for viscoelastic supplementation of joints in osteoarthritis, for filling tissues, especially the dermis, as so-called “dermal filiers” and for treating inflammatory diseases of the skin and mucous membranes. For example, WO 2005/067944 describes the treatment and prophylaxis of diseases of the skin and mucous membranes caused by herpes and papilloma viruses.

Surprisingly, it has now been found that glycosaminoglycan derivatives in which one or more amino groups are substituted by hydroxymethyl groups have an effect against enveloped viruses such as coronaviruses, RS viruses or influenza viruses.

The disaccharide units, from which glycosaminoglycans are built, consist, as mentioned above, of a uronic acid and an amino sugar. In the glycosaminoglycan derivatives according to the invention which are substituted by hydroxymethyl groups, a hydroxymethyl group is attached to one or more suitable reactive groups, for example nitrogen atoms of amino groups and / or hydroxyl groups. The glycosaminoglycans containing hydroxymethyl groups according to the invention thus have characteristic substituents such as -N (R) -CH ₂ OH, where R can be any radical, in particular H or acetyl, or -O-CH 2 OH. These are preferably bound to the aminosugars of the disaccharide units of the glycosaminoglycan. In the case of glycosaminoglycans which contain the amino sugar N-acetylglucosamine, the N-acetyl group is preferably substituted with a hydroxymethyl group. Characteristic of these hydroxymethyl groups-containing glycosaminoglycans within the meaning of the invention are -N (acetyl) -CH ₂ 0H- Groups. These compounds surprisingly show a particularly high anti-infective effect.

In the glycosaminoglycan derivatives according to the invention, one or more amino groups are substituted with hydroxymethyl groups. The degree of hydroxymethylation is preferably in the range from 200: 1 (0.5%) to 1: 1 (100%), preferably 100: 1 (1%) to 2: 1 (50%), particularly preferably 20: 1 ( 5%) to 10: 1 (10%) each on a molar basis. With increasing hydroxymethylation of the glycosaminoglycans, the virustatic and / or virucidal effect of glycosaminoglycans increases.

The glycosaminoglycans containing hydroxymethyl groups have excellent tissue compatibility. They remain in their place of action for a long time, and the length of stay can be controlled through the choice of the molecular weight of the glycosaminoglycan used and its degree of crosslinking. For example, depending on its molecular weight and degree of crosslinking, hydroxymethyl-hyaluronic acid remains at the site of action from about 30 minutes to six months.

Hyaluronic acid is basically biodegradable. It can have the same or different chain lengths or molecular weights, it can be short-chain (less than 100 repetitive units) or long-chain (over 100) with an influence on the duration of action. Cross-linking enables a further extension of the duration of action through reduced biodegradation. Hyaluronic acid is also a component of the bronchial secretion and is actively secreted by the surface cells of the lungs.

For the treatment of infectious diseases, glycosaminoglycans containing hydroxymethyl groups are suitable for the purposes of the invention both in uncrosslinked and in crosslinked form or as mixtures. Uncrosslinked or crosslinked hyaluronic acid or mixtures thereof are particularly preferably used.

Uncrosslinked glycosaminoglycans are preferably selected from (i) long-chain glycosaminoglycans with an average molecular weight (weight average) of at least 200 kD and (ii) short-chain glycosaminoglycans with an average molecular weight (weight average) of up to 50 kD or mixtures thereof. Crosslinked glycosaminoglycans can, for example, be covalently or non-covalently crosslinked. The crosslinked glycosaminoglycans can be prepared in a known manner. The covalent crosslinking generally takes place by crosslinking with bifunctional reactive agents, such as, for example, diepoxyoctane, BDDE, divinyl sulfone, glutaraldehyde or carbodiimide, via bifunctional amino acids, for example lysine, protamine or albumin. However, it is also possible, for example, to produce crosslinks via an amide, ester or ether bond. Further suitable reagents for the covalent crosslinking of glycosaminoglycans are ethylene glycol or 1-4-butanediol diglycidether, divinyl sulfone, photocrosslinking reagents such as ethyl eosin, hydrazides such as bishydrazide, trishydrazide and polyvalent hydrazide compounds. Furthermore, glycosaminoglycans which are intra- and / or intermolecularly esterified or crosslinked with hexamethylenediamine can also be used. Autocatalytic processes or non-covalent cross-linking using polyvalent metal ions such as iron, copper, zinc, calcium, magnesium, manganese, barium and other chelating metal ions are particularly preferred.

In use, the molecular weight is important, and in the case of crosslinked glycosaminoglycans, the degree of crosslinking, which is, for example, in the range from 0.1% to 10% on a molar basis, without being limited thereto. In general, it can be stated that with long-chain glycosaminoglycans a lower degree of crosslinking is sufficient to obtain a gel-like matrix, while with short-chain glycosaminoglycans a higher degree of crosslinking is required in order to obtain comparable properties.

The administration of the hydroxymethyl donor, e.g. the glycosaminoglycan containing hydroxymethyl groups, can in principle take place in any manner, provided that it is suitable for the treatment of the respective disease. Systemic or local administration is conceivable. In many cases, local administration takes place in the area of the diseased part of the body.

A hydroxymethyl group donor, for example a containing glycosaminoglycan, may be in the form of a pharmaceutical composition for use in the treatment of viral infections in which one or more hydroxymethyl group donors, for example containing hydroxymethyl groups Glycosaminoglycans are contained in an amount of preferably 0.01 to 20 percent by weight, based on the total pharmaceutical composition, in particular in an amount of 0.01 to 5 percent by weight and particularly preferably in an amount of 0.01 to 1 percent by weight.

The pharmaceutical compositions according to the invention can contain as pharmaceutical auxiliaries, e.g. agents for pH adjustment, stabilizers, antioxidants, solubilizers, penetration-promoting agents, preservatives and / and gelling agents, as are usually used in such compositions. They are used in the amounts customary in such preparations.

The combination of the active ingredient according to the invention with other active ingredients (e.g. corticoids, sympathomimetics, parasympathicolytics and / or leukotriene receptor antagonists) is a special embodiment and aims at the co-treatment of simultaneously existing non-viral respiratory diseases, such as bronchial asthma. A combination with sodium citrate can also be used to treat coagulopathy.

Also of importance are additives such as divalent or trivalent metal ions, which can have a crosslinking and stabilizing effect through chelation and which, on the other hand, can also accelerate the breakdown of the active substances.

In the tissue, glycosaminoglycans are naturally degraded by a large number of different enzymes or by oxygen radicals. Hyaluronic acid is broken down by hyaluronidases or oxygen radicals. Therefore, additives that inhibit enzymes such as hyaluronidase (heparin, indomethacin and / or salicylates) and those that prevent oxidative degradation in the tissue as so-called radical scavengers (vitamins A, E and / or C) are also of importance.

In a particularly preferred embodiment of the invention, a mixture of long-chain glycosaminoglycans (> 200 kD) with short-chain glycosaminoglycans (for example dimers, trimers, tetramers, pentamers or hexamers of the repetitive disaccharide units or larger units up to 50 kD) or mixtures of the aforementioned with crosslinked glycosaminoglycans are also used. Another particularly preferred one Embodiments are mixtures of cross-linked and non-cross-linked glycosaminoglycans.

The preparations according to the invention are preferably applied to the surfaces of the respiratory tract, e.g. nose, mouth, throat, windpipe, bronchi and / or lungs. If the mucous membranes of the respiratory tract are affected by an infectious disease, treatment using an aerosol as an inhalation solution can take place. The aerosols can be present in different droplet sizes from e.g. 0.5 micrometers to 100 micrometers, preferably from 5 to 30 micrometers. Methods for generating suitable aerosols for use in the respiratory tract are known. The propellant gas can be oxygen for the treatment of simultaneous shortness of breath. Suitable carrier media for the active ingredient according to the invention are physiological saline solutions (NaCl, Ringer's solution, acetate solutions), polyalcohols (glycerol), lipid emulsions.

Other application preparations in the form of a spray such as nasal sprays, gels that can be introduced into the mouth, nose and throat area, possibly also chewing gum, lozenges, bite capsules that release the active ingredient in the oral cavity, rinsing or drip solutions, e.g. eye drops, are also conceivable. Intravenous, systemic use is also possible.

By administering the preparations according to the invention it is possible to prevent or slow down the establishment of viruses, e.g. coronaviruses, in the airways, e.g. in the upper airways including the mouth and throat. Even with existing infections, the spread of the virus into the deeper airways or the lungs can be prevented or slowed down. In particular, by inactivating the viruses in the respiratory tract of an infected person, the transmission of contagious pathogens to healthy people can be prevented. In addition, virions which have already penetrated the organism and are inactivated by contact with the preparation according to the invention - in the sense of a vaccination with attenuated pathogens - can trigger an immune reaction which leads to the formation of specific antibodies.

The present invention preferably relates to the administration of the composition to human patients who already have a viral infection, for example a coronavirus infection. It is particularly preferably administered to Patients in whom the infection is at an early stage, especially at a stage in which the throat and possibly the upper airways, but not the lungs, are affected.

In the case of a coronavirus infection, the upper respiratory tract is first affected. There, the virus is already replicating in the epithelial cells. This stage lasts about 1 week. When testing, the result of the throat swab is positive. The infection then migrates into the deep respiratory tract and creates what is actually life-threatening viral pneumonia. If the agent according to the invention succeeds in initially containing the multiplication of the coronavirus, the exacerbation of the disease can be prevented, or the time window for developing immunity can be increased. Infection and contamination of healthy people can also be prevented.

It is also known that coronaviruses can also affect the gastrointestinal tract of humans. Consequently, the treatment principle described or the preparations described can be administered orally with the aid of a suitable carrier. A special form of preparation is a hydroxymethyl-modified, highly cross-linked hyaluronic acid gel or a hydroxymethyl-modified polysaccharide such as starch and cellulose.

The treatment principles mentioned are also used in veterinary medicine.

A composition according to the invention can be produced in a generally known manner which is customary per se for the production of such compositions. The order in which the individual components are mixed is generally not critical.

The type, dose and frequency of administration of the composition according to the invention and the nature (eg viscosity, degree of crosslinking, active ingredient content, etc.) depend in particular on the type and severity of the disease and on the age of the patient and the location and type of application. The type of treatment and the frequency of application depends in particular on the individual response of the person to be treated. Solutions are preferably applied one to several times a day.

The invention also includes mixtures of a glycosaminoglycan containing hydroxymethyl groups with other glycosaminoglycans in crosslinked and / and non-crosslinked form. Mixtures of hyaluronic acid containing hydroxymethyl groups and heparin are preferred. Mixtures of hyaluronic acid containing hydroxymethyl groups and positively charged glycosaminoglycans such as chitosamine are also preferred.

A glycosaminoglycan containing hydroxymethyl groups is produced as described in WO 2012/168462 and preferably comprises the steps:

(i) providing glycosaminoglycan and

(ii) Substituting one or more amino groups of the glycosaminoglycan with

Hydroxymethyl groups.

Hyaluronic acid is preferably used as the glycosaminoglycan starting product in step (i).

In step (ii) the glycosaminoglycan is then substituted with hydroxymethyl groups by chemical treatment. Step (ii) can be carried out together with step (i) or afterwards. Step (ii) can comprise, for example, the reaction of the glycosaminoglycan with formaldehyde or an agent which releases formaldehyde under the reaction conditions, such as taurolidine.

The glycosaminoglycan modified with hydroxymethyl groups can then be purified in a further step (iii). Excess formaldehyde or residues of formaldehyde-releasing reagents from step (ii) are removed. The purification can take place, for example, by precipitation with, for example, alcohols or salts, by chromatography processes, dialysis processes, vacuum extraction and / or freeze-drying.

A further step (iv) is then optionally carried out, in which the glycosaminoglycan substituted with hydroxymethyl groups is crosslinked. In principle, the crosslinking can also be carried out first and then the introduction of hydroxymethyl groups. The crosslinking can, as described above, take place according to methods known in the prior art.

In a further preferred embodiment of the invention, a glycosaminoglycan containing hydroxymethyl groups is then combined with one or more further active ingredients and / or additives. Examples of such further active ingredients and additives are explained above.

Another aspect of the invention relates to a combination of glycosaminoglycan containing hydroxymethyl groups with taurolidine, for example with a taurolidine solution, for example a 0.01 to 1.0% (w / v), in particular a 0.01 to 0.5% ( w / v) or a 0.1 to 0.2% (w / v) taurolidine solution. Hyaluronic acid is preferably used as the glycosaminoglycan, the molecular weight of the hyaluronic acid being, for example, between 100,000 and 10,000,000 Daltons.

The active ingredient according to the invention should be stored in a closed container, since hydroxymethyl residues can escape as formaldehyde via an equilibrium reaction in aqueous solution and the effectiveness is thus lost.

A preparation of cross-linked hyaluronic acid, which is isolated from the Hahnenkamm by means of formaldehyde (trade name Lubravisc; Fa. Bohus, Sweden), is approved in the EU for the treatment of degenerative joint diseases in humans and animals. Although the production with formaldehyde inevitably leads to a modification of the hyaluronic acid and the agent thus has an anti-inflammatory, and therefore medicinal, effect by blocking serine proteases in the joint, it is registered as a medical product.

Hyaluronic acid is isolated from streptococci without a formaldehyde step. The end product is free of hydroxymethyl groups, so an additional hydroxymethylation step must be carried out in order to achieve an antiviral effect.

Should the approval authorities for the present invention make an analogous decision, an accelerated approval process could be expected will. This would be desirable to contain the impending corona pandemic.

Another aspect of the invention relates to a pharmaceutical preparation for oral or nasal administration, in particular for administration as an aerosol or nasal spray, which contains a hydroxymethyl group donor as an active ingredient, optionally in a pharmaceutically acceptable carrier, advantageously in a powdery carrier or a liquid, e.g. an aqueous carrier such as physiological saline solution. This preparation is intended in particular for use in the respiratory tract and can in particular be used for one of the previously described medical indications including infection with SARS-CoV-2 or for the prophylaxis of infection with SARS-CoV-2.

In a preferred embodiment, the pharmaceutical preparation contains taurolidine as active ingredient, in particular in a concentration of 0.01 to 1.0% (w / v), in particular of 0.01 to 0.5% (w / v) or of 0, 1 to 0.2% (w / v) in a suitable, for example aqueous, carrier.

In a further preferred embodiment, the pharmaceutical preparation contains hydroxymethyl-modified hyaluronic acid, in particular in a concentration of 0.01 to 1.0% (w / v), in particular 0.01 to 0.5% (w / v) or of 0.1 to 0.2% (w / v) in a suitable vehicle, for example an aqueous vehicle.

Another aspect of the invention relates to a pharmaceutical preparation for oral administration, in particular for administration as a gel, tablet or capsule, which contains a hydroxymethyl group donor as an active ingredient, optionally in a pharmaceutically acceptable carrier, advantageously in a solid carrier or in a liquid carrier, for example a aqueous carrier such as physiological saline. This preparation is intended in particular for use in the digestive tract and can in particular be used for one of the medical indications described above including an infection with SARS-CoV-2 or for the prophylaxis of an infection with SARS-CoV-2. This preparation is optionally provided with a coating, for example a gastric acid-resistant coating, so that the active ingredient can be released in the intestine, for example in the duodenum, jejunum and / or colon. In a preferred embodiment, the pharmaceutical preparation contains a hydroxymethyl-modified hyaluronic acid, for example a hydroxymethyl-modified crosslinked hyaluronic acid gel or a modified polysaccharide such as starch and cellulose in a suitable carrier, for example a solid or a liquid carrier.

To improve compatibility or to reduce undesirable effects and to control the pharmacological effect of the active substance, the aqueous carriers can have a physiological osmolality and buffered pH values, preferably in the physiological range, for example between 6.0 and 8.0.

Another object of the invention relates to the use of a hydroxymethyl group donor or a pharmaceutical preparation as described above for reducing and / or avoiding side effects of a vaccination against a viral infection, in particular an infection with coronaviruses such as SARS-Cov-2. The vaccination is preferably a vaccination with a genetic vaccine, i.e. a vaccine that contains a nucleic acid coding for a viral antigen as an active ingredient, particularly preferably with a genetic vaccine against a coronavirus such as SARS-CoV-2. The genetic vaccine is particularly preferably an mRNA or a vector active substance, in particular a vaccine which contains the genetic information for a coronavirus spike protein.

For this purpose, the hydroxymethyl group donor or a pharmaceutical preparation which contains the hydroxymethyl group donor as an active ingredient is administered to a person to be vaccinated or vaccinated. It can be administered prophylactically together with the vaccination, preferably at an interval of about 24 hours or less, in particular about 12 hours or less, from the vaccination, or, alternatively, therapeutically after the onset of symptoms. Administration can be carried out by any of the above-mentioned methods, with inhalation and intraperitoneal, e.g. intravenous, administration being preferred. Taurolidine is preferably used as a hydroxymethyl group donor.

Another object of the invention relates to an in vitro method for inactivating viruses, in particular enveloped viruses such as coronaviruses, e.g. SARS-CoV-2 viruses, paramyxoviruses, e.g. RS viruses, or orthomyxoviruses, e.g. influenza viruses, comprising the treatment of a preparation of enveloped viruses with a Hydroxymethyl group donor under conditions which lead to a killing and / or attenuation of the viruses in the treated preparation.

The treatment preferably reduces the cytopathic effect (CPE) of the viruses in the treated preparation by a factor of 10, 10 ² , 10 ³ , 10 ⁴ , 10 ⁵ , 10 ⁶ or more. Suitable standard tests are available to the person skilled in the art for determining the CPE of viruses. The CPE of SARS-CoV-2 can be determined, for example, with a test as described by K. Gorshkov et al. "The SARS-CoV-2 Cytopathic Effect is Blocked by Lysosome Alkalizing Small Molecules", ACS Infect Dis. (December 2020): acsinfectdis.0c00349. The inactivation step according to the invention can optionally be combined with further inactivation steps such as heating, acid treatment and / or irradiation, for example with UV or ionizing rays.

The inactivation step is usually carried out by treating the virus preparation in an aqueous medium to which the flydroxymethyl group donor is added. The medium can optionally also contain organic, water-miscible solvents such as DMSO. The final concentration of the hydroxymethyl group donor in the preparation can be selected, for example, as described above. If necessary, however, the hydroxymethyl group donor can also be used in a higher concentration. When using taurolidine, for example, concentrations of 0.01-10% (w / v) or 0.1-3% (w / v) are suitable. The treatment is advantageously carried out at a temperature in the range from about 5 ° C to about 50 ° C. The duration of the treatment is usually at least 5 minutes, preferably at least 10 minutes.

The virus preparation treated with a hydroxymethyl group donor can optionally be used as a vaccine, which can be administered in a suitable manner, for example by injection and / or by inhalation in the form of an aerosol.

Yet another object of the invention is an inactivated preparation of viruses, in particular enveloped viruses such as coronaviruses, e.g. SARS-CoV-2 viruses, paramxyoviruses, e.g. RS viruses, or orthomyxoviruses, e.g. influenza viruses, which are labeled with a hydroxymethyl group Donor has been modified. The cytopathic activity of the treated virus preparation is favorably reduced by at least a factor of 10, 10 ² , 10 ³ , 10 ⁴ , 10 ⁵ or 10 ⁶ compared to an untreated virus preparation.

The inactivated virus preparation according to the invention can be used, for example, as a vaccine for human and / or veterinary medicine.

Pilot study

The clinical efficacy and the virustatic effect of taurolidine and hydroxymethyl-modified hyaluronic acid were checked on the basis of a pilot study with volunteer participants in terms of individual therapeutic treatment.

The study was carried out on a total of 19 patients with symptoms of a respiratory illness or a cold.

Of these, 6 tested positive and 4 negative for SARS-CoV-2 (point of care test, POCT). For the remaining participants, the clinical symptoms were the indication for initiation of therapy.

The taurolidine treatment was carried out by inhaling 1 - 2 ml aerosol with 0.5% taurolidine using an ultrasonic nebuliser or compressed air nebuliser. (According to the manufacturer, these devices generate aerosols with a predominant particle size of 5 micrometers). It was applied 2 to 4 times a day, depending on the symptoms. Taurolidine was taken from a commercial 2% stock solution (TauroNova from Tauropharm) and diluted with physiological saline solution or with a buffered saline infusion solution (pH 7.2).

Alternatively, a 0.5% solution with hydroxymethyl-modified hyaluronic acid was used as an aerosol for inhalation or as a nasal spray. The approved commercial product Lubravisc with a hyaluronic acid content of 2% was diluted to 0.5% with physiological saline solution. The hyaluronic acid modification was achieved by isolating it from the cockscomb with the help of formaldehyde. There is a transfer of hydroxymethyl groups to the hyaluronic acid.

The time interval from the start of treatment until the symptoms subsided was measured in each case. There was no hospitalization for the participants. example 1

A 64-year-old male participant received a positive SARS-CoV-2 antigen test. Ten days later he fell ill with a fever of over 38 ° C, muscle pain and a feeling of weakness. He then inhaled 0.5% taurolidine twice a day. 4 days later, the body temperature and well-being normalized.

Example 2

A 32-year-old participant had close family contact with a person who was proven to have Covid-19. When symptoms of illness such as a rise in body temperature, a feeling of weakness and pain in the back muscles arose shortly afterwards, the POCT became positive. Accordingly, he began inhaling taurolidine 0.5% 3 times a day. The complaints were resolved within 48 hours by the time the patient was rehabilitated.

Example 3

A 52-year-old participant developed a cough, burning eyes, fever and lung pain. She tested positive for Covid-19 with POCT. She then inhaled taurolidine 0.5% 3 times a day. On day 4 after starting therapy, she was symptom-free.

Example 4

Two participants, a couple, male and female, 72 and 69 years old, respectively, became infected with a close 96-year-old relative who tested positive for SARS-CoV-2 and were taken into quarantine after the test results were also positive. During this time they both developed dry, tickly coughs and started inhaling taurolidine 0.5% 3 times a day. The cough stopped in both participants after 3 days. There were no other symptoms of the disease.

Example 5

A 13-year-old participant lived in close domestic contact with the sick mother who tested positive for SARS-Cov-2. He developed a sore throat and tested positive with POCT as a result. He then inhaled taurolidine 0.5% twice a day. On the 4th day he was symptom-free. Example 6

2 female and 2 male participants between the ages of 27 and 64, members of a family, who had close contact with two relatives who had tested positive and who suffered from Covid-19, inhaled taurolidine 0.5% 3 times as a prophylactic measure, if their relatives were aware of their disease daily or twice a day for 7 days, simultaneously with the infected relatives. The 4 participants remained symptom-free and the POCT for SARS-CoV-2 remained negative in all.

Example 7

A 29-year-old participant developed a sore throat with an unspecific malaise. He inhaled modified hyaluronic acid twice a day. From the 3rd day he was symptom-free. There was no testing for SARS-CoV-2.

Example 8

One participant developed a runny nose and a sore throat. She used modified hyaluronic acid as a nasal spray twice a day. The symptoms subsided on the 3rd day after the start of therapy.

Example 9

2 participants with cold symptoms such as runny nose, cough, sore throat inhaled taurolidine 0.5% 3 times a day immediately after the symptoms appeared. They were symptom-free 3 to 5 days after the start of therapy. A SARS-CoV-2 test was not carried out.

Example 10

A 28-year-old participant suffered from cold symptoms such as cough, sore throat and subfebrile body temperatures for 4 days. She inhaled taurolidine 0.5% once on the evening of the 4th day and was symptom-free the next day. A test for SARS-CoV-2 was not carried out.

Claims

Expectations

1. Hydroxymethyl group donor for use in the treatment or prevention of coronavirus, paramyxovirus or orthomyxovirus infection.

2. Hydroxymethyl group donor according to claim 1, characterized in that it is selected from glycosaminoglycans containing hydroxymethyl groups, in which one or more amino groups and / or hydroxyl groups are substituted with hydroxymethyl, or is selected from taurolidine

3. Hydroxymethyl group donor according to claim 2, characterized in that the glycosaminoglycan is selected from hyaluronic acid, heparin, chondroitin sulfate, chitosamine and poly-N-acetyl-glucosamine, preferably hyaluronic acid in which one or more amino groups are substituted with hydroxymethyl.

4. hydroxymethyl group donor according to claim 2 or 3, characterized in that the glycosaminoglycan is selected from hyaluronic acid.

5. Hydroxymethyl group donor according to one of claims 2 to 4, characterized in that the degree of hydroxymethylation in the range from 200: 1 (0.5%) to 1: 1 (100%), preferably 100: 1 (1%) up to 10: 1 (10%).

6. hydroxymethyl donor according to any one of claims 2 to 5, characterized in that the hydroxymethyl group-containing

Glycosaminoglycan is present in uncrosslinked form, the

Glycosaminoglycan in particular is selected from

(i) long chain glycosaminoglycans with an average

Molecular weight (weight average) of at least 200 kD and

(ii) short chain glycosaminoglycans with an average

Weight average molecular weight up to 50 kD or mixtures thereof.

7. Hydroxymethyl group donor according to one of claims 2 to 5, characterized in that the hydroxymethyl group-containing glycosaminoglycan is in crosslinked form, the degree of crosslinking being in particular in the range from 0.1% to 10%.

8. hydroxymethyl group donor according to any one of claims 2 to 7, characterized in that the hydroxymethyl group-containing glycosaminoglycan is present as a mixture of uncrosslinked and crosslinked glycosaminoglycans.

9. hydroxymethyl donor according to claim 1 or 2, characterized in that it is taurolidine.

10. Hydroxymethyl group donor according to one of the preceding claims, characterized in that it is present in combination with at least one inhibitor of glycosaminoglycan degradation, the inhibitor of glycosaminoglycan degradation being selected in particular from heparin, Indomethacin, salicylates, free radical scavengers such as vitamin A, C or E, and mixtures thereof.

11. Hydroxymethyl group donor according to one of the preceding claims for use in human medicine or veterinary medicine.

12. hydroxymethyl group donor according to any one of the preceding claims, for local administration.

13. Hydroxymethyl group donor according to one of the preceding claims for administration to surfaces of the upper respiratory tract.

14. Hydroxymethyl group donor according to one of the preceding claims, characterized in that it is present as an inhalable preparation, in particular as an aerosol, or as a nasal spray.

15. Hydroxymethyl group donor according to one of the preceding claims for administration to patients with an infection of the upper and / or deep respiratory tract.

16. Hydroxymethyl group donor according to one of the preceding claims for the treatment of an infection with coronaviruses, in particular for the treatment of patients with an infection with SARS-CoV 2.

17. A method for the treatment or prevention of an infection with coronaviruses, paramyxoviruses or influenza viruses, characterized in that a subject to be treated, in particular a human subject, is given Administered preparation which contains a hydroxymethyl group donor as defined in any one of claims 1 to 10, in an amount sufficient to treat the disease.

18. Hydroxymethyl group donor for use as an active ingredient for the treatment and prevention of an inflammatory disease of the respiratory tract, for example COPD, ARDS or cystic fibrosis, in particular of an inflammatory disease of the respiratory tract associated with a viral infection.

19. A method for the treatment or prevention of an inflammatory disease of the respiratory tract, for example COPD, ARDS or cystic fibrosis, in particular of an inflammatory disease of the respiratory tract associated with a viral infection, characterized in that a subject to be treated, in particular a human subject, is given Administered preparation which contains a hydroxymethyl group donor as defined in any one of claims 1 to 10, in an amount sufficient to treat the disease.

20. Pharmaceutical preparation for oral or nasal administration, in particular for administration as an aerosol or nasal spray, characterized in that it contains a hydroxymethyl group donor as an active ingredient in a pharmaceutically acceptable carrier.

21. Pharmaceutical preparation according to claim 20, characterized in that it contains taurolidine, in particular in a concentration from 0.01 to 1.0% (w / v), in particular from 0.01 to 0.5% (w / v) or from 0 , 1 to 0.2% (w / v) in an aqueous vehicle.

22. Pharmaceutical preparation according to claim 20, characterized in that it contains hydroxymethyl-modified hyaluronic acid, in particular in a concentration of 0.01 to 1.0% (w / v), in particular from 0.01 to 0.5% (w / v) or from 0.1 to 0.2% (w / v) in an aqueous carrier.

23. Pharmaceutical preparation according to one of claims 20-22 for use in the respiratory tract.

24. Pharmaceutical preparation for oral administration, in particular for administration as a gel, tablet or capsule, characterized in that it contains a hydroxymethyl group donor as an active ingredient in a pharmaceutically acceptable carrier.

25. Pharmaceutical preparation according to claim 24, characterized in that it is provided with a coating, for example a gastric acid-resistant coating.

26. Pharmaceutical preparation according to claim 24 or 25, characterized in that it contains a hydroxymethyl-modified hyaluronic acid, e.g. a hydroxymethyl-modified crosslinked hyaluronic acid gel or a modified polysaccharide such as starch and cellulose in a solid or a liquid carrier.

27. Pharmaceutical preparation according to one of claims 24-26 for use in the digestive tract.

28. Hydroxymethyl group donor or pharmaceutical preparation which contains a hydroxymethyl group donor as an active ingredient in a method for reducing and / or avoiding side effects of a vaccination against a viral infection.

29. Hydroxymethyl group donor or pharmaceutical preparation which contains a hydroxymethyl group donor as active ingredient, for use according to claim 28, in a vaccination against coronaviruses, in particular with SARS-CoV-2.

30. Hydroxymethyl group donor or pharmaceutical preparation which contains a hydroxymethyl group donor as active ingredient, for use according to claim 28 or 29, in a vaccination with a genetic vaccine, in particular with a genetic vaccine against a coronavirus such as SARS-CoV-2.

31. Hydroxymethyl group donor or pharmaceutical preparation which contains a hydroxymethyl group donor as active ingredient, for use according to one of claims 28-30 in a vaccination with an mRNA or a vector active ingredient.

32. Hydroxymethyl group donor or pharmaceutical preparation which contains a hydroxymethyl group donor as active ingredient, for use according to one of claims 28-31 in a vaccination with a vaccine which contains the genetic information for a coronavirus spike protein.

33. Hydroxymethyl group donor or pharmaceutical preparation which contains a hydroxymethyl group donor as an active ingredient, for use according to one of claims 28-32 for intraperitoneal, for example intravenous, or inhalative administration.

34. In vitro method for inactivating viruses, in particular enveloped viruses such as coronaviruses, e.g. SARS-CoV-2 viruses, paramyxoviruses, e.g. RS viruses, or orthomyxoviruses, e.g. influenza viruses, comprising treating a preparation of viruses, in particular enveloped ones Viruses with a hydroxymethyl group donor under conditions which lead to a killing and / or attenuation of the viruses in the treated preparation.

35. Virus preparation, in particular of enveloped viruses such as coronaviruses, for example SARS-CoV-2 viruses, paramyxoviruses, for example RS viruses, or orthomyxoviruses, for example influenza viruses, characterized in that they contain viruses killed and / or attenuated with a hydroxymethyl group donor .

36. Use of a virus preparation produced according to claim 34 or a virus preparation according to claim 35 as a vaccine.