PROCEDURE AND EQUIPMENT FOR NO INSTILLATION

Description

Subject of the invention

[0001] The present invention relates to a multi-stage dilution process for producing a solution containing nitric oxide. The invention further relates to a device for carrying out the dilution process according to the invention and to an instillation unit comprising this device. The invention also relates to the device and the instillation unit in their use for treating diseases, in particular diabetic-related circulatory disorders and chronic wounds. Background of the invention

[0002] Numerous processes and devices for the production of nitrogen monoxide (NO) are known from the state of the art.

[0003] According to EP 1 903 003 A1, NO can be produced by photolysis of a photolabile NO precursor. The reaction proceeds in the presence of radical scavengers and antioxidants to form highly pure NO. When applied to NO production within liquids, this process generally only results in a slow increase in the NO concentration.

[0004] According to WO2013 / 063354, an NO-releasing foot bath can be produced by adding a polysiloxane polymer derivatized with diazeniumdiolate groups to the bath solution. This polymer then reacts with water to form NO. Since NO generation occurs through spontaneous decomposition of the polymer side groups, the release kinetics can only be inadequately controlled. Furthermore, this process requires a considerable amount of time to build up a therapeutically relevant NO level.

[0005] US 2016 / 051579 A1 relates to a method for treating bovine respiratory diseases with nitric oxide and teaches, in Example 2 (

[0280] and

[0281] ), the acidification of a sodium nitrite solution with citrate as a solid to produce an NO-releasing solution. 900 µl of the resulting NO-releasing solution are mixed with 100 µl of a bacterial preparation, and the resulting solution is then serially diluted.

[0006] US 2013 / 0028942 A1 concerns the production of aerosols containing NO and nitrite, based on a solution that is pre-acidified with citric acid (Example 1,

[0381] ).

[0007] US 2015 / 0157657 A1 concerns the production and provision of solutions with sustained NO release. In US 657, an aqueous solution of NaNO2 is introduced, acidified with solid citric acid, and rapidly mixed.

[0008] There is therefore still a need for new processes for the production of NO-containing solutions in which NO can be produced quickly and yet in a controlled manner with high purity.

[0009] The object of the invention is therefore to provide a process for the production of therapeutically usable NO solutions which is improved with respect to at least one of the above-mentioned disadvantages. Summary of the invention

[0010] This object is achieved according to the invention by providing a process for producing a nitrogen monoxide (NO)-containing solution, which comprises the following steps: (a) Providing a first reaction component comprising a pH-labile NO donor; (b) Providing a second reaction component which is an aqueous solution having a pH between 2.5 and 7.0; (c) Mixing the first and second reaction components to form an aqueous solution having a pH between 3.0 and 5.5 and generating NO to form an NO-containing aqueous solution; (d) Diluting the NO-containing aqueous solution from step (c) by mixing with an aqueous dilution solution to result in a dilution factor of 5 to 20, and the aqueous dilution solution is a buffered solution or distilled water, so that the pH of the mixture from step (c) is increased by at least one pH level.

[0011] Specific embodiments of the invention are the subject of further dependent and independent claims.

[0012] The method according to the invention combines several decisive advantages over the methods known from the prior art.

[0013] Surprisingly, this method was found to meet the complementary requirements of NO release kinetics. Thus, a suitable, concentrated stock solution of NO can be rapidly built up in the aqueous solution in an acidic environment, which then undergoes a pH increase through mixing with an aqueous diluent. This allows the NO concentration to be stabilized over a longer period of time and thus maintained in a controlled manner.

[0014] By storing the first and second reaction components separately, the NO-containing stock solution can be provided in a precisely controlled and demand-based manner.

[0015] The short half-life of NO typically hampers its therapeutic use. With the method according to the invention, despite the short half-life, the NO level can be maintained for a sufficient period of time by stabilizing the NO in the weakly acidic, neutral, or basic carrier medium, while simultaneously achieving a therapeutically relevant concentration.

[0016] Due to the presence of antioxidants, the process allows the production of NO in a purity required for therapeutic or cosmetic applications.

[0017] Numerous pH-labile NO donors are known from the state of the art, such as nitrite salts, NONOates or nitrosothiols, which the person skilled in the art can use here.

[0018] Due to the highly controlled release, the process can be used in devices that release only very small amounts of NO. This is a crucial advantage, especially for NO as a highly potent bioactive molecule. Furthermore, it allows the development of a corresponding device (such as a wound dressing or a foot bath) as a medical device (e.g., a so-called Class III medical device), provided that in these specific cases the device's effect is primarily determined by its mechanical or physical properties.

[0019] By simply adapting the procedure with regard to NO donors and acids, it can be specifically adapted to the treatment requirements.

[0020] The process according to the invention also makes it possible to dispense with the external supply of NO.

[0021] The method according to the invention is a simple method using mostly known substances, so that it can be carried out quickly and inexpensively, as well as being easy to use in therapy with a low susceptibility to errors.

[0022] The multi-stage dilution process is particularly well suited for automating the manufacturing process due to the separate provision of reaction components, which are stable and non-toxic in this form.

[0023] Due to the aforementioned advantages, the method is particularly suitable for use in negative pressure wound therapy, so that the wound can be rinsed with an NO-containing rinsing solution and then, after suctioning off the solution, exposed to a therapeutically advantageous negative pressure.

[0024] The devices used in the manufacturing process open up further scope for freedom with regard to the characteristic parameters and the choice of materials.

[0025] In summary, the NO production process according to the invention enables NO-based therapies in which the highly reactive and correspondingly unstable gas NO can be administered in a controlled manner in a cost-effective, reliable, safe and user-customizable manner. The invention in detail

[0026] The invention thus includes a multi-stage process in which NO generation is first induced in an acidic environment by mixing the first and second reaction components and, after a selected period of time, the pH is then increased by diluting this mixture with an aqueous dilution solution in order to stop or reduce the pH-dependent NO synthesis and to provide an NO-containing aqueous solution with a therapeutically applicable NO concentration.

[0027] By increasing the pH to the preferably slightly acidic, neutral, or basic range, the regeneration of toxic NO2 radicals is prevented. The presence of at least one antioxidant according to the invention eliminates NO2 radicals and other radicals produced during NO generation, so that the aqueous liquid is enriched with highly pure NO.

[0028] The starting point of the process is the first reaction component, which comprises a pH-labile donor and preferably also contains an antioxidant.

[0029] NO generation is initiated by mixing this first reaction component with the second reaction component. The second reaction component is an aqueous solution with a pH between 2.5 and 7.0. By adding the first reaction component, which comprises the pH-labile NO donor, NO generation is initiated by providing or creating an acidic environment. It is noteworthy that the second reaction component can also have a weakly acidic or neutral pH, which as such is not suitable for inducing NO production. However, in addition to the pH-labile donor, the first reaction component can also contain an acidic substance (such as citric acid), which, when mixed with the aqueous second reaction component, results in a correspondingly acidic pH.

[0030] For cosmetic or medical applications, the mixture must also contain at least one antioxidant at the point at which an acidic pH value allows NO generation. For this purpose, the at least one antioxidant can already be present in the first reaction component with the pH-labile NO donor and added with it to the second reaction component. In the following, NO donor and antioxidant are also referred to as substance classes.

[0031] In one embodiment, the first reaction component is a mixture comprising a pH-labile NO donor and at least one antioxidant. In a preferred embodiment, the pH-labile donor and the at least one antioxidant are present in separate compartments in the first reaction component, so that they are only present in one compartment after being mixed together with the second reaction component. In a particularly preferred embodiment, the nitrite as the pH-labile NO donor and the ascorbic acid or a salt thereof (i.e., an ascorbate) are present in separate compartments in the first reaction component.

[0032] The separate compartments can be formed by a vessel or area with a dividing wall or membrane. Alternatively, one or even both substance classes, i.e., NO donor and antioxidant, can be present in encapsulated form (e.g., as a microcapsule), so that the capsule wall acts as a separation barrier. Even if the two encapsulated substance classes, or the encapsulated and non-encapsulated substance classes, were mixed, they would still be present in separate compartments.

[0033] Alternatively, the at least one antioxidant can also be present in the aqueous, acidic solution of the second reaction component. This has the advantage that the components contained in the carrier medium are protected from unwanted oxidation during production and / or storage by the presence of at least one antioxidant. This can be particularly advantageous for devices according to the process, such as wound dressings or plasters, since the addition of additional substances is difficult here and these must have sufficient storage stability.

[0034] According to the invention, the first reaction component comprises, as an essential component, a pH-labile NO donor, i.e., a substance capable of generating NO in an acidic aqueous environment. This first reaction component is preferably in the form of a solid, foam, gel, cream, or liquid. Particularly preferably, the first reaction component is in the form of a solid, which is particularly preferably a powdered or granular solid.

[0035] According to the invention, an aqueous solution having a pH of 2.5 to 7.0 is used as the second reaction component. Mixing with the first reaction component produces an aqueous solution having a pH in the range of 3.0 to 5.5, preferably in the range of 3.5 to 5.3, and particularly preferably in the range of 4.0 to 5.0.

[0036] According to the invention, an aqueous dilution solution is used to dilute the aqueous NO-containing solution. Any aqueous solution that is physiologically safe and in which the NO is soluble at the therapeutically relevant concentration can be used. According to the invention, the dilution solution is a buffered solution or distilled water.

[0037] According to the invention, a pH-labile NO precursor (also called NO donor (NOD)) is used for NO generation. pH-labile NO donors are known in the art and familiar to those skilled in the art.

[0038] In a preferred embodiment of the invention, the pH-labile NO donors are selected from the group comprising organic nitrates, inorganic nitrates, inorganic nitrites, organic nitrite esters such as alkyl nitrites, sulfur, nitrogen or oxygen nitroso compounds, NO-metal compounds and NO-chelating substances.

[0039] Examples of pH-labile NODs include inorganic nitrites, alkyl nitrites such as isopentyl nitrite, diazeniumdiolate derivatives, trans[RuCl(

[15] aneN4)NO] 2+< , 6-nitrobenzo[a]pyrole, S-nitroso-glutathione, S-nitroso-thiol, S-nitroso-N-acetyl-D-penicillamine (SNAP), nitroaniline derivatives, 2-methyl-2-nitrosopropane, imidazoyl derivatives, nitrate esters, hydroxylnitrosamine, hydroxylamine, hydroxyurea and sodium nitroprusside.

[0040] Preferably, the pH-labile NO donor is an inorganic nitrite salt, which is advantageously a pharmacologically acceptable substance. Examples of such nitrites include alkali or alkaline earth metal nitrites. Examples include: LiNO2, NaNO2, KNO2, RbNO2, CsNO2, FrNO2, Be(NO2)2, Mg(NO2)2, Ca(NO2)2, Sr(NO2)2, Ba(NO2)2, or Ra(NO2)2, and combinations thereof.

[0041] Particularly preferred NOD is NaNO 2 , which is further preferably used together with a combination of ascorbic acid and / or Trolox as antioxidants in the first reaction component or in the reaction mixture.

[0042] The concentration of the nitrite salt or nitrite salts in the aqueous solution as a mixture of first and second reaction components is preferably between 1 and 100 mM, particularly preferably between 5 and 70 mM and in particular between 10 and 50 mM.

[0043] In an alternative embodiment, a nitrate salt can also be used, which can be enzymatically converted into the corresponding nitrite salt. Nitrates of alkali or alkaline earth metals are preferred. Examples include: LiNO 3 , NaNO 3 , KNO 3 , RbNO 3 , CsNO 3 , FrNO 3 , Be(NO 2 ) 3 , Mg(NO 2 ) 3 , Ca(NO 2 ) 3 , Sr(NO 2 ) 3 , Ba(NO 2 ) 3 , or Ra(NO 2 ) 3 .

[0044] In order to remove the multiply oxidized nitrogen oxides, oxygen radical anions, or hydroxyl radicals that occur during NO generation, it is necessary for cosmetic or medical applications that at least one antioxidant is present during the reaction. Therefore, in the process according to the invention, the first reaction component and / or the second reaction component and / or the dilution solution contains at least one antioxidant.

[0045] Preferably, only the first reaction component contains the at least one antioxidant. This at least one antioxidant is then dissolved together with the pH-labile NO donor in the aqueous solution as the second reaction component and can thus exert its antioxidant effect directly at the beginning of NO generation.

[0046] Depending on the type of chemical mechanism of action, antioxidants are divided into radical scavengers or reducing agents.

[0047] In oxidation reactions between organic compounds, chain-like radical transfers often occur. Substances with sterically hindered phenol groups are involved, forming inert, stable radicals during these transfers that do not react further, thus terminating the reaction cascade (radical scavengers). These include natural substances such as tocopherols and synthetic ones such as butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), and gallates.

[0048] Furthermore, reducing agents with a very low standard redox potential of less than +0.4 V (at pH 7.0 and 25°C) can also be used. Typical examples include ascorbic acid (-0.04 V at pH 7 and 25°C), salts of sulfurous acid (+0.12 V at pH 7 and 25°C), and certain organic sulfur-containing compounds (e.g., glutathione, cysteine, thiolactic acid).

[0049] In a preferred embodiment, the at least one antioxidant is capable of reducing the HNO 2 present as an NO donor in an acidic environment to NO. For this purpose, the antioxidant as a reducing agent must have a standard redox potential of less than +1.0362 volts, preferably less than +0.5 volts, particularly preferably less than +0.2 volts, and especially preferably less than 0 volts.

[0050] The at least one antioxidant is advantageously capable of reducing the harmful NO 2 radical to the NO 2 anion. For effective elimination of the NO 2 radical, the at least one antioxidant should preferably have a bimolecular reaction constant k that is greater than 1.0 x 10 6 < M -1 < s -1 < and preferably greater than 1.0 x 10 7 < M -1 < s -1 <. Antioxidants suitable according to the invention with the associated reaction constants are disclosed in Kirsch et al., 2002 (Biol. Chem 383; 389-399, see Table 1). Examples include: captopril thiolate, caffeic acid, sinapic acid, ferulic acid, lycopene, zeaxanthin, lutein, astaxanthin, canthaxanthin, arachidonate, Gly-Tyr dipeptide, tyrosine, purines and pyrimidines such as the nucleobases adenine, guanine, cytosine, thymine, uracil and the corresponding derivatives and analogues thereof, including the nucleosides and nucleotides containing them.

[0051] In a further embodiment, the aqueous solution used according to the invention comprises, in addition to the antioxidant, an antioxidant synergist. Synergists support the effect of antioxidants, for example, by regenerating depleted antioxidants (so-called "redox cycling"). By complexing trace metals (sodium EDTA) or creating an oxidation-inhibiting pH, synergists can enhance the antioxidant effect of a radical scavenger or reducing agent. Typical examples of antioxidant synergists are EDTA, 1-hydroxyethane-1,1-diphosphonic acid, citric acid, fumaric acid, uric acid, and 2-(hydroxymethyl)-1,4-benzyldiol.

[0052] In the production process according to the invention, ascorbate or ascorbic acid is particularly preferably used as an antioxidant.

[0053] Particularly suitable for the aqueous solutions according to the invention are water-soluble vitamin E derivatives such as Trolox or alpha-AMG, organic sulfur-containing compounds such as glutathione, cysteine, or thiolactic acid or also organic acids such as ascorbic acid, alpha-lipoic acid, hydroxycinnamic acids such as p-coumaric acid, ferulic acid, sinapic acid or caffeic acid, or hydroxybenzoic acids such as gallic acid, procatechuic acid, syringic acid or vanillic acid.

[0054] Other preferred antioxidants include polyphenolic compounds such as anthocyanins, flavonoids and phytoestrogens.

[0055] For the aqueous reaction solution, water-soluble representatives of the aforementioned groups can be conveniently combined, for example ascorbate and (RS)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), ascorbate and cysteine, or preferably ascorbate and N-acetylcysteine.

[0056] Advantageously, the at least one antioxidant is present in a molar excess relative to the NO donor.

[0057] In one embodiment of the invention, the second reaction component in its configuration as an aqueous liquid additionally contains one or more of the following substances: catalysts, detergents, buffer substances, chromophores, substances that stabilize the prodrug such as dimethyl sulfoxide or ethanol, substances that increase the half-life of NO, as disclosed, for example, in US 2003 / 0039697, NOD stabilizers, antioxidants, dyes, pH indicators, care substances, fragrances, pharmacologically active substances.

[0058] The skilled person will select suitable substances or mixtures of substances based on their general technical knowledge and the respective intended use. In doing so, they will particularly consider that physiologically compatible and / or dermatologically compatible substances and mixtures are used when using the reaction component for topical application. Acid activation in step (c)

[0059] To cleave the pH-labile NO donor, the first reaction component is mixed with the second reaction component to produce a sufficiently acidic pH. According to the invention, this pH is low enough to induce the cleavage of the pH-labile NO donor to form NO. The specific pH depends on the pH instability of the NO donor and the desired time span for NO generation. The lower the pH, the faster NO will be generated in the reaction mixture. After NO generation, this reaction mixture serves as the NO-containing stock solution for the subsequent dilution step (step d).

[0060] According to the invention, the pH in step (c) is between 3.0 and 5.5, preferably between 3.5 and 5.2, particularly preferably between 4.0 and 5.0, and in particular 5.0. As already explained above, the optimal pH depends on the particular NO donor used and the intended reaction rate and can be adjusted accordingly by a person skilled in the art.

[0061] Preferably, the process step of NO generation, measured from the mixing of the first and second reaction components, has a duration of between one minute and 60 minutes, preferably between 15 minutes and 45 minutes and particularly preferably from 20 to 30 minutes.

[0062] The NO-containing aqueous stock solution formed by the reaction is diluted with an aqueous dilution solution by a factor between 5 and 20, preferably between 8 and 12 and particularly preferably by a factor of 10, so that the pH of the NO-containing stock solution is increased by at least one pH level.

[0063] Advantageously, the solution resulting from the dilution step (d) has a NO concentration of between 10 µM and 1000 µM, preferably between 20 µM and 500 µM and particularly preferably between 75 µM and 200 µM. Pharmacologically active substances

[0064] In one embodiment of the invention, one of the reaction components additionally contains one or more pharmacologically active substances. These can support the pharmacological action of NO or act independently of NO in a manner therapeutically relevant for the respective disease.

[0065] Examples of pharmacologically active substances include: anti-inflammatory drugs such as nonsteroidal anti-inflammatory drugs (NSAIDs) or corticoids, immunosuppressants, antibiotics, anticoagulants, antithrombotics, antiviral agents, antifungals, local anesthetics and analgesics. device

[0066] It is a further object of the present invention to provide an improved device for producing a therapeutically usable NO-containing aqueous solution.

[0067] In a further aspect, the invention provides an apparatus for carrying out the method according to the invention, which comprises at least three containers as follows: (a) a first container for holding the first reaction component, which is connected to a second container via an opening or conduit, (b) a second container for holding the liquid second reaction component, which container is further connected to a third container via a further opening or conduit for the passage of liquid; and (c) a third container for holding the dilution liquid, wherein the line or opening between the individual containers is closed by a shut-off element that can be opened or removed to establish communication between the individual containers, and wherein the first container is a closure cap pre-filled with the first reaction component, which closes the second container and has a trigger mechanism which, when actuated, allows mixing of the first reaction component in the closure cap and the second reaction component in the second container.

[0068] In a preferred embodiment, the trigger mechanism is a push button or the lid is designed as a rotatable lid with a thread. With the rotatable lid, a piercing element (e.g. a spike-shaped or blade-shaped element) attached to the underside of the lid can pierce the separation membrane upon rotation and the downward movement of the closure lid resulting from the thread, thus enabling mixing of the two reaction components. Accordingly, when pressed, the push button can pierce the separation membrane by a piercing element attached to the underside of the lid, thus leading to mixing. Advantageously, the first container, which is preferably a pre-filled closure lid, as well as the partition wall, are made of a chemically inert material.

[0069] In a particularly preferred form, the closure cap comprises one or more compartments in which individual substance classes such as pH-labile NO donor and antioxidant can be accommodated separately from one another.

[0070] In a further embodiment, the device, and in this case the second container, is provided with a mixing device that ensures rapid and complete mixing of the reaction components with the aqueous solution. In a preferred embodiment, this mixing device operates within the closed system of the device, without requiring direct intervention in the device and, in turn, in the containers; for example, no stirring rod should have to be inserted from the outside. This can be achieved, for example, by a magnetic stirring element. A magnetic stirring bar (so-called "stirring fish") can be used, which is driven by a magnetic stirring device located adjacent to, but outside, the device, which generates a rotating magnetic field.

[0071] Another example is a holder for containers 1 and 2, which, for example, transmits vibrations to them in order to achieve a release of the reaction components.

[0072] In a preferred embodiment, this mixing device is therefore controlled and / or driven in a controlled manner by a stirring device located outside the device, which is preferably part of the installation unit.

[0073] In a preferred embodiment, the second container and / or the third container is designed as a plastically deformable plastic bag. This enables further strategies for simple and non-invasive mixing of the reaction components in the aqueous solution.

[0074] Numerous plastic materials are available to the person skilled in the art for this purpose, these being preferably selected from the group comprising polypropylene, polyethylene, ethylene vinyl acetate (EVA), polyvinyl chloride (PVC), polyethylene terephthalate (PET), oriented polypropylene (OPP), biaxially oriented polypropylene (BOPP), oriented polyamide (OPA) and biaxially oriented polyamide (BOPA).

[0075] In another embodiment, the bag consists of a multilayer polyolefin-based film. Examples include: polyethylene / ethyl vinyl alcohol copolymer / polyethylene (PE / EVOH / PE), PP / EVOH / PP, PP / EVOH / PE, polyamide / polyethylene (PA / PE), PE / PA / PE, PP / PA / PE, PA / EVOH / PA / PE and PP / PA / EVOH / PA / PE, EVOH / OPP, EVOH / BOPP, EVOH / OPA, EVOH / BOPA, and PVDC / PET.

[0076] Thus, the unit receiving the bag, preferably the instillation unit, can comprise a support plate onto which the bag is placed, wherein the support plate is movable and is designed such that it can perform a horizontal rotation, a rocking, a vibration or a shaking movement.

[0077] In an alternative embodiment, the unit accommodating the bag comprises a pressure device that can exert pressure on the bag. This pressure is preferably applied only to a portion of the bag, so that the liquid is forced into adjacent areas of the bag. Mixing occurs after the pressure is released or removed by redistributing the liquid.

[0078] The expert has numerous options for designing such a printing device. Examples include: The pressure device can be designed as a stamp that laterally compresses the preferably flat bag and mixes the bag contents by alternately raising and lowering the stamp. The pressure device can be a roller that partially compresses the bag and mixes the bag contents by a reciprocating movement. The pressure device can be an eccentric element that periodically exerts pressure on the bag via an asymmetrically arranged rotation axis.

[0079] In a further embodiment, the device and / or the unit accommodating the device comprises a transport device for transporting the liquid from the second container to the third container. Examples of the following transport devices are mentioned here, which can be used individually or in combination.

[0080] In a first embodiment, the third container is under negative pressure, so that when the valve located between the second and third containers is opened, the liquid is drawn into the third container. This can be easily achieved by a third container that is under a form tension, for example, a plastic container made of a duromer that has been emptied beyond its fill volume, resulting in a "dent."

[0081] In a second embodiment, an excess pressure is built up in the second container, which forces the liquid into the third container. This can occur, for example, endogenously through a chemical reaction (e.g., by adding a carbonate salt to the first reaction component, which leads to the formation of CO2 in the acidic environment of the liquid in the second container). Alternatively, the second container can also have a gas connection through which gas is introduced into the second container from the outside, thereby forcing the liquid into the third container. The position of the connection and the position of the containers are the same as stated above for the first embodiment.

[0082] In a third embodiment, the mixing device according to the invention also functions as a transport device. For example, the roller acting on the bag could transfer the liquid into the third bag when the valve between these containers is opened and a movement is made toward the third container, squeezing the bag out.

[0083] In a fourth embodiment, the opening or line provided between the second and third containers has a connection option for a pumping device. Thus, with a hose line, the liquid can be pumped from the second container to the third container by a hose pump, which is preferably designed as a peristaltic pump.

[0084] In a preferred fifth embodiment, the first container has a vent connection at its upper end, ie at the end opposite the connection point to the third container, so that when this connection is opened and the second bag is stored vertically above the third bag, air can penetrate into the second bag and the liquid can flow downwards into the third bag.

[0085] It is a further object of the present invention to provide an improved instillation unit for delivering a therapeutically usable NO-containing aqueous solution.

[0086] In a further aspect, the invention provides an instillation unit which comprises the device according to the invention and is selected from the group comprising wound dressing, wound plaster, spray, inhaler, bathing device and shower device.

[0087] In one embodiment, the instillation unit is constructed to include the three containers of the device as an integral, i.e., permanently mounted, component.

[0088] In an alternative embodiment, the instillation unit is designed to allow reversible coupling or receptacle to one or more containers.

[0089] The instillation unit may, for example, have a cavity that accommodates the three containers and connects the third container to the instillation unit for dispensing the finished NO-containing solution.

[0090] Preferably, the instillation unit comprises one or more of the following components: (a) a control system for opening and closing the shut-off elements between the device containers; (b) a monitoring system for controlling the opening status of the shut-off elements; (c) a pumping device for transporting or circulating the aqueous solutions; (d) a stirring system for mixing the liquids in one or more of the containers of the device; (e) a temperature control unit for controlling the temperature of the reaction components and / or the dilution solution and / or the diluted NO-containing solution; (f) a computing and storage unit for controlling the components of the instillation unit. Instillation unit with shower device

[0091] In one embodiment, the instillation unit comprises a shower device.

[0092] Such a shower device reduces the risk of (re-)contamination of wounds by microbes from, for example, neighboring skin areas, since the contaminated aqueous solution flows directly away from the skin area and is replaced by new, non-contaminated aqueous solution.

[0093] Unlike an immersion bath, the skin is not overly softened. Furthermore, since there's no need to fill an immersion container, this method of application is also faster, and treatment can begin immediately after generating the active ingredient-containing bath solution.

[0094] For unstable active ingredients, such as NO, shower application allows for the easier preparation of bath solutions with a constant NO concentration.

[0095] A shower device allows for more flexible application, allowing the treatment to be focused on the necessary areas of the body.

[0096] In one embodiment of the invention, the shower device comprises a plurality of spaced-apart shower heads, which are preferably connected to one another via a common liquid line.

[0097] In a further embodiment, a switching device is provided on the shower head, by means of which different types of jets can be generated with the shower head, for example a normal water jet and a shower jet.

[0098] In a special embodiment, the shower device generates a pulsating water jet which, as a massage jet, additionally supports the therapeutic effect of vasodilating agents, such as NO.

[0099] In a special embodiment, the shower head utilizes the Venturi nozzle principle and enables the mixing of a bathing solution containing no active ingredients with a bathing solution containing active ingredients. Preferably, the container for the bathing solution containing active ingredients, which is integrated into the shower head, is connected to the water supply. Through the connection to the Venturi nozzle located on the container, the bathing solution containing active ingredients is transported from the container and passed on in this mixture to the shower head holes. Mixing occurs, for example, by activating a switch that opens the connection between the Venturi nozzle and the bathing solution container containing active ingredients.

[0100] The shower device is advantageously designed to prevent the release of the active ingredient, particularly gaseous active ingredients such as NO, from the bathing solution into the air. For this purpose, the shower head can, for example, have an air intake at the edge so that the NO escaping from the water jets is immediately extracted and either returned to the bathing solution in the shower head or removed from the system (e.g., by filtration, adsorption, or degradation).

[0101] This can also be achieved with a shower head with two different outlet areas: a first, inner area of ​​the shower head is intended for the active ingredient-containing bathing solution, and a second, annular outlet area, surrounding the inner area, is intended for a non-active ingredient bathing solution. This second area forms a "shell" of non-active ingredient bathing solution and ensures that the active ingredient escaping from the bathing solution in the first area is dissolved within it and does not enter the environment.

[0102] In a further embodiment of the invention, the shower device is not designed as a shower head, but as a hose or pipe with outlet openings, wherein the hose or pipe is preferably shaped as a ring or spiral. In a preferred embodiment, the ring or spiral is attached to the inner wall of a shower chamber of the shower device, with the outlet holes facing inward.

[0103] In a preferred embodiment, the shower device is designed such that it can be placed on or attached to the body part to be treated, thereby preferably building up a film of water that runs off the body part. This design has the advantage that it requires particularly small amounts of bathing solution containing active ingredients, and the film formation (unlike a spray device) particularly effectively prevents the release of potentially toxic active ingredients into the environment. For this design, the hose or the (half) ring can be guided partially or completely around the body part to be treated and locked in place, for example, by a slight clamping effect. In an alternative design, the shower device can also be designed as a bracket whose shape is adapted to the body part to be treated.

[0104] In a special embodiment, a shower curtain is attached to the aforementioned ring, hose, or bracket. When these shower devices are attached to the body, this close-fitting shower curtain further prevents the release of the active ingredient.

[0105] In a further embodiment, the shower device is designed as a drainage stocking, bandage or glove and thus allows a targeted release of the active ingredient to the body.

[0106] Furthermore, the shower device can be combined with one or more body covers, so that only the area to be treated is accessible for the shower application. In a preferred embodiment, this cover can have one or more recesses for the body area to be treated.

[0107] In a further embodiment, the outlet opening of the shower device is designed in a slit shape, allowing the shower device to function as a waterfall shower. Compared to a shower head with many individual water jets, a waterfall shower results in a smaller release of the active ingredient into the environment.

[0108] The shower device is conveniently equipped with a switch that regulates the water supply.

[0109] In addition, the shower device can also have a pressure regulator that regulates the water pressure and thus the amount of water coming out. Instillation unit in combination with the NPWT wound dressing

[0110] In a preferred embodiment, the instillation unit is used in conjunction with a wound dressing. For this purpose, the installation unit can be coupled to a wound dressing in such a way that the wound can be rinsed with the NO-containing solution. The NO-containing solution produced by the device according to the method of the invention is transported through a line to the wound dressing, where it is introduced into the space between the wound and the wound dressing, and then rinses the wound. The use of a peristaltic pump is preferred here, as it allows a pumping process without contaminating the pump.

[0111] The wound dressing is conveniently equipped with a suction device so that the solution can be sucked out again after the wound has been rinsed.

[0112] Preferably, this suction device is designed to create a negative pressure in the wound space beneath the wound dressing, which is particularly preferably between -60 and -200 mm Hg. In particular, the suction device is capable of creating a negative pressure of -60, -70, -80, -85, -90, -100, -110, -120, -130, -140, -150, -160, -170, -180, -190, or -200 mm Hg. computer program product

[0113] In one aspect, the application discloses a computer program product which can be loaded directly into a storage unit and comprises software sections with which the method according to one of the claims can be carried out when the computer program product is executed on an installation unit. Therapeutic or cosmetic use

[0114] In a particular aspect, the invention thus provides a device for carrying out the method according to the invention, which is suitable for use in the treatment or prevention of diseases, wherein the patient is exposed to the NO released from the device.

[0115] This treatment is preferably performed externally or topically. For example, by applying an NO-releasing patch or wound dressing to a specific area of ​​skin requiring treatment, that area can be targeted.

[0116] Alternatively, in a bathing device, the affected body part can be treated by immersion in the NO-containing liquid or by spraying, pouring or pouring the NO-containing liquid over it.

[0117] The device according to the invention can be used in particular to stimulate the metabolism of tissues through external application, in the field of dermatology for the treatment of surgical or accident-related wounds, chronic, non-healing or poorly healing and / or bacterial or fungal wounds, as well as for the treatment of dermatological diseases from the group of inflammatory, immunologically controlled or autoimmune diseases.

[0118] In a preferred embodiment, the disease treated with the device according to the invention is selected from the group comprising neuropathic pain, varicose veins, ischemia and thrombopathic diseases, allergies, skin infections, skin inflammations, atopic dermatitis, in particular neurodermatitis, dermatomyositis and pemphigus vulgaris; wound defects, such as chronic diabetic-neuropathic ulcers, leg ulcers, decubitus wounds;primary healing wounds, secondary healing infected wounds, burns, hidradenitis supparativa (acne inversa), warts, diaper rash, razor burn, complications of skin grafting, erectile dysfunction, angina pectoris, heart failure, left heart failure, coronary heart disease, anginal symptoms after myocardial infarction, anal fissure, spasms of the smooth muscles of the esophagus, menstrual problems, Reynaud's syndrome, Buerger's syndrome, peripheral arterial disease (PAD), peripheral arterial occlusive disease (PAD), inflammatory and autoimmune diseases of the skin (psoriasis, dermatitis, neurodermatitis), fungal diseases of the skin, bacterial, mycotic and parasitic diseases of the skin (e.g. leishmaniasis), tinea cruris and tinea inguinalis.;

[0119] Solutions prepared using the method according to the invention can preferably be used in the form of an inhalation spray for the treatment of obstructive lung diseases. Furthermore, they can be used to induce local vasodilation of constricted or occluded blood vessels. In this case, it is preferred to administer the solution directly into the heart, for example, by an endoscopic procedure.

[0120] In one embodiment, the device according to the invention can be used to treat local circulatory diseases in animals, such as laminitis in horses, and generally veterinary diseases that correspond to or are similar to the human diseases listed here.

[0121] The device according to the invention can also be used to treat muscular dystrophy (MD). Treatable forms of MD include: Duchenne MD, Becker-Kiener MD, Emery-Dreifuss MD type 1, scapulopereonal MD, reducing body myopathy (RBM), limb-girdle dystrophies, congenital muscular dystrophies, distal muscular dystrophies, vocal cord and pharyngeal weakness with distal myopathy (VCPDM), myofibrillar myopathies, and myotonic dystrophies.

[0122] An inflammation treatable with the device according to the invention can be a bacterial, viral, mycotic, or parasitic infection. The bacterial infection can be caused, for example, by a bacterium selected from the group consisting of S. aureus, B. circulans, B. cereus, E. coli, P. vulgaris, P. acnes, S. pyogenes, S. enterica, V. anguillarum, K. pneumoniae, P. piscicida, P. aeruginosa, A. tumefaciens, M tuberculosis, and M ulcerans. The fungal infection can be caused by a fungus selected from the group containing T. equinum, C. albicans, F. oxysporum, R. solani, B. cinerea, and A. ylavus.The fungal infection to be treated may involve the skin or nails, as in onychomycosis. Viral infections may be caused by one of the following virus families: Poxviridae, Rotavirus, Papillomavirus, Parvovirus, and Varicellavirus. The NO-releasing device can preferably be used to treat skin infections in which the virus Molluscum contagiosum The parasitic infection can be caused, for example, by a parasite of the following genera: Plasmodium, Leishmania, Schistosoma, Austrobilharzia, Heterobilharzia, Ornithobilharzia, or Cryptosporidium. Of particular note here is the pathogen Plasmodium falciparum.

[0123] In one embodiment, the device according to the invention can be used to treat the paroxysmal circulatory disorders (sickle cell crises) that occur in sickle cell anemia. The active ingredient used in such cases, hydroxyurea, is believed to inhibit the formation of the deoxygenated T variant in erythrocytes, thus preventing the transformation to the sickle cell phenotype. By binding the released NO to hemoglobin, however, the non-sickle cell-forming R variant is formed, which can be associated with improved blood flow and even the prevention of sickle cell crises.

[0124] In a further embodiment, the device according to the invention can be used to treat hair loss, and in particular androgenetic alopecia. The treatment here includes both slowing or stopping hair loss and even regrowth of hair. Other forms of hair loss that can be treated according to the invention include alopecia praematura, alopecia areata, alopecia areata atrophicans, alopecia totalis, alopecia universalis, diffuse alopecia, alopecia actinica, alopecia mechanis such as alopecia liminaris, alopecia marginalis frontalis traumatica, alopecia seborrhoica, alopecia muciosa, and alopecia parvimaculata. Analogous to the mode of action of the drug minoxidil, the NO should result in an increased supply of blood, oxygen, and nutrients to the hair follicles through increased blood flow to the scalp.

[0125] According to the invention, the device can be used, for example, as follows: 1.) on open wounds, since it has surprisingly been found that the application according to the invention does not cause skin irritation; 2.) for MRSA prophylaxis in high-risk patients; or 3.) as a synergistic application with conventional antibiotics, since it has surprisingly been found that, as a result of NO action, the conventional antibiotics can effectively combat the remaining inflammation.

[0126] In a preferred embodiment, the device according to the invention is used for the treatment of chronic wounds of the lower extremities of diabetics. Furthermore, the prophylactic treatment can reduce the risk of developing chronic wounds and the number of medical amputations. Thus, the reduction of neuropathic leg pain and the creation of an improved wound environment are associated with a noticeably improved quality of life for the patients. Furthermore, a significant reduction in treatment costs can be expected due to the shortened wound care time.

[0127] In addition, it may be possible to address systemic diseases such as high blood pressure (hypertension) and related hemodynamic disorders by treating larger areas of the body.

[0128] In one embodiment of the invention, the device according to the invention is used to treat poorly healing wounds. Impaired arterial circulation and / or impaired venous return are significant causes in the development and chronicity of wounds of the lower extremities. NO-induced arterial vasodilation improves circulation to the affected tissue, and the antithrombogenic effect of NO significantly promotes or facilitates venous return of blood. The NO-dependent improvement in both hemodynamic parameters represents the crucial therapy-relevant aspect of a local and systemic effect that significantly reduces the risk of wound formation and significantly accelerates their healing. The NO delivered to the body part to be treated using the device according to the invention can therefore be successfully used to treat poorly healing wounds.

[0129] In a particular embodiment, the device according to the invention is used to treat diabetic pain of the lower extremities, i.e., the foot and / or leg. Diabetic pain is a very common occurrence in the course of diabetes. Diabetic foot / leg pain is a result of long-term elevated blood glucose concentrations, which is the root cause of the nerve and vascular damage observed during diabetes. NO-induced arterial vasodilation improves blood flow to the affected tissue and helps influence pain transmission, thereby reducing pain. The NO delivered externally to the foot and / or leg using the device according to the invention can therefore be successfully used to treat diabetic foot / leg pain.

[0130] In a specific embodiment of the invention, the device according to the invention is used to treat patients with (skin) transplants, and in particular to treat poorly perfused flaps. The two previously mentioned hemodynamic variables, arterial blood flow and venous return, are also essential parameters for the success of surgical flap surgery. Flap surgery refers to surgical plastic surgical techniques that transfer skin and / or tissue from a (dispensable) part of the same individual to a new desired location. As a rule, these are pure skin flaps, but any tissue with or without skin can be transplanted, either with a pedicle (i.e. with its associated blood-supplying vessels and nerves) or freely (i.e. with the blood vessels connected to the blood supply of the new area).The functional acceptance of the transplanted tissue depends exclusively on the arterial blood supply and regulated venous outflow. NO-induced arterial vasodilation improves blood flow and thus the necessary supply to the flap, while the antithrombogenic effect of NO promotes and facilitates venous outflow and return of blood. Externally administered NO preparations can therefore ensure or promote the success of a flap-based treatment option.

[0131] In a further embodiment, the invention also provides a cosmetic method in which the NO produced by the method or device according to the invention acts on the skin of a human. DEFINITIONS

[0132] For the purposes of the invention, a "reaction component" is defined as a single substance or a mixture of substances which require mixing with another reaction component in order to react.

[0133] According to the invention, the term "treatment" is understood to mean any application of the device according to the invention to an individual which serves to alleviate the disease symptomatically or causally or even to suppress it completely or to stop, delay or postpone the progression of the disease.

[0134] According to the invention, "instillation" is understood to mean the administration of liquid pharmaceutical or other medicinal compositions onto body surfaces or into the organism for therapeutic, diagnostic, or preventive purposes. Accordingly, an instillation unit is a device that administers the liquid pharmaceutical composition.

[0135] In contrast to infusion, where the medication is distributed throughout the body via the circulatory system (systemic therapy), with instillation the medication remains at the site of application and acts locally.

[0136] In the context of the present invention, "prevention" refers to the avoidance of the occurrence of diseases, particularly vascular or metabolic diseases, and thus the reduction of their prevalence and the mitigation of their impact on morbidity and mortality in the population. The central strategy is to suppress or completely eliminate the trigger factors of diseases.

[0137] Prevention includes primordial prevention, primary prevention, secondary prevention, tertiary prevention and quaternary prevention.

[0138] Primary prevention begins before the onset of the disease and aims to prevent a recurrence of the disease. Primary prevention is aimed at at-risk groups, healthy individuals, and those without symptoms.

[0139] Primary prevention can be distinguished from primordial prevention, which begins even earlier. It aims to prevent the occurrence of risk factors.

[0140] Secondary prevention begins in the early stages of a disease. It serves the purpose of early detection and containment of disease progression or chronicity. Often, the pathogenetic process has already begun at this point, without any noticeable symptoms. The target group are individuals who participate in the preventive measure while healthy or asymptomatic, but who become patients as a result of the diagnostic procedure.

[0141] Tertiary prevention takes place after acute treatment or the onset of a disease. It aims to prevent subsequent complications and relapses. It is aimed at patients with chronic impairments and those undergoing rehabilitation. One example is the prevention of recurrences in cancer.

[0142] There is also quaternary prevention, which aims to prevent unnecessary medicine or overdoses and takes into account the principle of “primum non nocere” as a cornerstone of all medicine.

[0143] Terms used in the claims such as "comprise," "have," "include," "contain," and the like do not exclude further elements or steps. The use of the indefinite article does not exclude a plurality. A single device may perform the functions of multiple units or devices recited in the claims. Reference numerals indicated in the claims are not to be construed as limitations on the means and steps employed.

[0144] In accordance with the foregoing description, the following embodiments are disclosed, which alone or in any combination with the aforementioned embodiments are part of the invention. EXAMPLES Example 1. pH-induced NO production process 1.1 Materials:

[0145] Device for the quantification of NO: Eco physics CLD 822 Reaction chamber: quartz glass, approx. 100x100x10mm (approx. 100ml volume) Buffer solution: 150 mM acetic acid, 150 mM NaOH in distilled water Base: 1M NaOH Sodium L-ascorbate 1M NaNO 2 1.2 Experimental procedure

[0146] 0.56 g of sodium L-ascorbate was dissolved in 98.6 ml of buffer solution, transferred to the reaction chamber, and 1.4 ml of NaNO2 (1M) was added. The sodium nitrite concentration was 14 mM, and the ascorbate concentration was 28.3 mM. The final solution measured a pH of 5.0.

[0147] Over a period of 60 min, a 200 µl sample was taken at intervals of 2-3 minutes and the NO content was quantified using the CLD system. 1.3 Results

[0148] The results of the NO measurements as a function of the reaction time are shown in Figure 1A continuous increase in the NO concentration is observed, reaching a value corresponding to a concentration of 1.11 mM in the liquid after 60 minutes. FIGURE LEGENDS

[0149] The invention is explained in more detail below with reference to the figures, without limiting the invention to them. They show: Fig. 1: At pH 5.0 and a suitable buffer system, an NO concentration of 1 mM is reached in the concentrate (A) after approximately 30 minutes. Dilution with distilled water (e.g., 1:10) leads to an increased pH (e.g., 6.0) with a stable NO concentration (e.g., 100 µM). Fig. 2: A schematic representation of a device 1 for carrying out the dilution process with a first container 10 containing the first reaction component 15 in solid form (preferably sodium nitrite and sodium ascorbate). This first reaction component 15 can be mixed with the second reaction component 25, which is held in the second container 20, by pressing (black arrow) the push button 11 and piercing the separation membrane 13 with the piercing element 12. The second container 20 is fluid-connected to the third container 40 via a valve 30.After opening the valve 30, the acidic NO-containing solution in the container 20 can be diluted by the dilution solution as the third reaction component 45 and simultaneously raised in pH. Fig. 3: A schematic representation of a device 1 for carrying out the dilution process according to . Figure 2 , where the result of the push-button operation is shown. The pressed push-button 11 has pierced the separation membrane 13 with the piercing element 12 and thus connected the first container 10 and the second container 20 to one another in such a way that a mixture 26 of the first and second reaction components has now been achieved in them. Fig. 4: A schematic representation of a device 1 for carrying out the dilution process according to Figure 2, wherein nitrite particles 16 as pH-labile NO donor and ascorbate particles 17 as antioxidant are present in the first container 10 in two separate compartments. Fig. 5: A schematic representation of a device 1 for carrying out the dilution process according to Figure 4, wherein the result of the push-button operation is shown. The pressed-in push-button 11, together with the piercing element 12, has pierced the separating membrane 13 and thus connected the first container 10 and the second container 20 to one another in such a way that a mixture 26 of the nitrite particles and the ascorbate particles with the liquid second reaction component has now been achieved in them. Fig. 6 in A to D shows four different mixing devices for the second container 20, designed as a bag, with the pre-filled push-button 10, 11 as the first container, with a support plate 100 designed as a rocker plate 60 and rotation axis 61 in A; with an eccentric device 70 and a rotation axis 71 in B; with a laterally movable roller 80 in C and with an up-and-down movable plunger in D. REFERENCE SYMBOL

[0150] 1Device for carrying out the dilution process 10First container 11Push button 12Puncture element 13Separation membrane 15First reaction component 16Nitrite particles as the first component of the first reaction component 17Ascorbate particles as the second component of the first reaction component 20Second container 25Second reaction component 26Mixture of first and second reaction components 30Valve 40Third container 45Third reaction component 50Connection for withdrawing liquid from the third container 60Rocking plate (see arrows for rocking movement) 61Rotation axis for support plate 70Eccentric element 71Rotation axis for eccentric element 80Laterally movable roller (dashed rollers = extreme positions) 90Pushpin 100Support plate for bag

Claims

1. Method for preparing an aqueous solution containing nitrogen monoxide (NO), comprising the following steps: (a) providing a first reaction component comprising a pH-labile NO donor; (b) providing a second reaction component which is an aqueous solution having a pH between 2.5 and 7.0; (c) mixing the first and the second reaction component to form an aqueous solution having a pH between 3.0 and 5.5 and generating NO to form an aqueous solution containing NO; (d) diluting the aqueous solution containing NO from step (c) by mixing with an aqueous dilution solution to result in a dilution factor of 5 to 20, and the aqueous dilution solution is a buffered solution or distilled water, so that the pH of the mixture from step (c) is increased by at least one pH step.

2. Method according to claim 1, characterized in that the first reaction component is in the form of a solid, foam, cream, gel or liquid and preferably as a powdered or granular solid.

3. Method according to claim 1 or 2, characterized in that the pH-labile NO donor is selected from the group comprising inorganic nitrite salt, alkyl nitrites such as isopentyl nitrite, diazeniumdiolate derivatives, trans[RuCl([15]aneN4)NO]2+, 6-nitrobenzo[a]pyrrole, S-nitrosoglutathione, S-nitrosothiol, S-nitroso-N-acetyl-D-penicillamine (SNAP), nitroaniline derivatives, 2-methyl-2-nitrosopropane, imidazoyl derivatives, nitrate esters, hydroxyl-nitrosamine, hydroxylamine, hydroxyurea, sodium nitroprusside, and is preferably an inorganic nitrite salt.

4. Method according to claims 1 to 3, characterized in that the pH-labile NO donor is selected from the group LiNO2, NaNO2, KNO2, RbNO2, CsNO2, FrNO2, Be(NO2)2, Mg(NO2)2, Ca(NO2)2, Sr(NO2)2, Ba(NO2)2, or Ra(NO2)2 and combinations thereof and is preferably NaNO2.

5. Method according to any of the preceding claims, characterized in that that the first reaction component and / or the second reaction component and / or the dilution solution contains at least one antioxidant, and preferably only the first reaction component contains the at least one antioxidant.

6. Method according to any of the preceding claims, characterized in that the at least one antioxidant is selected from the group comprising ascorbate and derivatives thereof, tocopherol, tocotrienol, tocomonoenol and derivatives thereof, butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), glutathione, cysteine, thiolactic acid, alpha-lipoic acid, p-coumaric acid, ferulic acid, sinapic acid, caffeic acid, gallic acid, procatechuic acid, syringic acid, vanillic acid, polyphenolic compounds from the group of anthocyanins, flavonoids or phytoestrogens, and is preferably a mixture of an ascorbic acid derivative and a tocopherol derivative.

7. Method according to any of the preceding claims, characterized in that method step (c) has a duration of between 1 minute and 60 minutes, preferably between 15 minutes and 45 minutes and particularly preferably between 20 and 30 minutes.

8. Method according to any of the preceding claims, characterized in that the aqueous solution resulting from the dilution in step (d) has a NO concentration of between 10 µM and 1000 µM, preferably between 20 µM and 500 µM and particularly preferably between 75 and 200 µM.

9. Device for carrying out a method according to any of claims 1 to 8, comprising at least three containers: (a) a first container for receiving the first reaction component, which is connected to a second container via an opening or line, (b) a second container for receiving the liquid second reaction component, wherein said container is further connected to a third container by a further opening or line for the passage of liquid; (c) a third container for receiving the dilution liquid; wherein the line or opening between the individual containers is closed by a shut-off element that can be opened or removed to establish connection between the individual containers, and wherein the first container is a closure cap pre-filled with the first reaction component, which closes off the second container and has a trigger mechanism which, when actuated, allows mixing of the first reaction component in the closure cap and the second reaction component in the second container.

10. Instillation unit comprising a device according to claim 9, characterized in that it is selected from the group comprising wound dressing, wound plaster, spray, inhaler, bathing device and shower device, wherein the instillation unit is preferably used for negative pressure therapy.

11. Instillation unit according to claim 10, characterized in that it also comprises: (a) control system for opening and closing the shut-off elements between the device containers; (b) optionally a monitoring system for monitoring the opening status of the shut-off elements; (c) pumping device for transporting or circulating the aqueous solutions; (d) optionally a stirring system for mixing the liquids in one or more of the containers of the device; (e) (optionally) a temperature control unit for controlling the temperature of the reaction components and / or the dilution solution and / or the diluted solution containing NO; (f) a computing and storage unit for controlling the components of the instillation unit.

12. Device according to claim 9 or instillation unit according to claim 10 or 11 for use in the treatment or prevention of diseases, characterized in that the patient's body surface or hollow organs are exposed to the liquid containing NO released from the device or instillation unit.

13. Cosmetic procedure comprising the action of liquid containing NO on the skin of a human, characterized in that a method according to claims 1 to 8, a device according to claim 9 or an instillation unit according to claim 10 or 11 is used, the device or the installation unit being used for producing the liquid containing NO used in the cosmetic procedure.