HK1107014A - Diaminooxidase-containing pharmaceutical compositions - Google Patents

Description

pharmaceutical composition containing diaminooxidase

The present invention relates to compositions for treating or preventing histamine-induced diseases and disorders.

Histamine (1H-imidazole-4-ethylamine) is formed by enzymatic decarboxylation of histidine and is therefore a basic biogenic amine with a molecular weight of 111 Da.

Histamine is virtually ubiquitous in the organism. Produced by the human body itself and stored in a passivated form in metachromatic granules of mast cells and basophils where they can be released immediately. The highest histamine concentration was measured in the lungs. After release, histamine is the most potent mediator of a large number of physiological and pathological processes, often also by interaction with cytokines.

Furthermore, histamine can also reach the body from the outside, on the one hand by inhalation or in the oral route, for example by ingestion of histamine-containing foods such as cheese, wine, canned fish and kimchi.

The most important functions and roles of histamine in the human or animal body are:

1) dilate capillary, improve capillary permeability and reduce blood pressure.

2) Contracting smooth muscles, especially the bronchial muscles in the lungs.

3) Inducing higher gastric acid secretion.

4) Improving the heart rhythm.

5) Histamine is a mediator of allergic emergency type reactions and is the most important mediator of allergic diseases such as allergic rhinitis (Heuschnupfen) and bronchial asthma.

6) Furthermore, histamine is a typical cause of urticaria (nestselausschlag) and plays an important role in drug allergy or incompatibility.

High concentrations of free circulating histamine cause undesirable effects such as headache, nasal difficulty or discharge, airway obstruction, tachycardia and extra systole, as well as gastric and intestinal discomfort, which can lead to soft stools up to diarrhea, and hypotension. Eyelid swelling is also often described, and occasionally urticaria. In addition, redness of the skin, a drop in blood pressure and bronchospasm may also occur. The following table shows the symptoms associated with histamine concentrations in blood.

Histamine (ng/ml)	Body reaction
		0-1	Is free of
1-2	Enhancement of gastric secretion
		3-5	Tachycardia and skin irritation
6-8	Drop in blood pressure
		7-12	Bronchospasm
About 100	Cardiac arrest

In mammalian organisms, histamine is decomposed by two enzymes: diamino oxidase (DAO, EC 1.4.3.6) and histamine-N-methyltransferase (NMT, EC 2.1.1.8) (Mizuguchi et al, 1994). DAO catalyzes the oxidative deamination of histamine to imidazaldehyde; NMT catalyzes N-methylation to form N-methyl histamine.

Two decomposition pathways are essential for the organism: DAO removes histamine, for example, ingested by food into the gastrointestinal tract, and NMT controls histamine signaling in the nervous system (Kitanaka et al, 2002).

The main task of DAO is to prevent histamine ingested from the intestine by food from reaching the blood circulation. This protective mechanism can also be allergic shock in extreme cases (Taylor 1986, Nilsson et al 1996). DAO is a secreted protein and therefore works extracellularly, whereas N-methyltransferases are only active in the cytosol (Kufner et al, 2001).

Natural DAO, in addition to histamine, can also decompose other biogenic amines such as putrescine, spermidine and cadaverine and is obtained, for example, from pig kidney. Native DAO is a copper-containing homodimeric glycoprotein in which subunits are linked by disulfide bridges. DAO has a molecular weight of about 182kDa (Kluetz and Schmidt 1997, Rinaldi et al, 1982) and a carbohydrate content of about 11% (Shahand Ali 1988). This enzyme belongs to the group of copper-containing aminooxidases which catalyze the oxidative deamination of primary amines to form aldehydes, ammonia and hydrogen peroxide according to the general reaction scheme below (Bachrach 1985): RCH₂NH₂+H₂O+O₂＝>RCHO+NH₃+H₂O₂Wherein the radical R contains an amino group.

Copper-containing aminooxidases are characterized by Topachinon in the active center, which is formed by post-translational modification of conserved tyrosine residues (James et al, 1990, James et al, 1992, Mu et al, 1992).

DAO is found primarily in the small intestine, liver, kidney and white blood cells in the blood. For pregnant women, DAO also forms in the placenta. Pregnant women have blood DAO levels about 500 to 1000 times higher than non-pregnant women. DAO is continuously produced and excreted into the intestinal lumen. Thus, in healthy persons, histamine-rich foods already release histamine to a large extent in the intestine. The retained histamine is decomposed by the DAO present therein when passing through the intestinal mucosa. Histamine is decomposed into imidazole acetaldehyde and further imidazole-acetoacetate. The cofactor of DAO is 6-hydroxydopa and possibly pyridoxal phosphate, vitamin B6. DAO is a sensitive enzyme that can be hindered by a variety of substances such as other biogenic amines, alcohols and their breakdown products acetaldehyde, and a variety of different drugs. DAO activity has not been demonstrated in neural tissue at present.

As mentioned above, exogenous histamine absorbed through food as well as endogenous histamine can also cause a wide variety of pathologies due to allergic reactions. In terms of clinical value, based on reduced DAO activity, it is highlighted by the intolerability of at least three forms of histamine:

a minority of people have an innate deficiency of DAO and are not lost either.

During infection of the intestinal mucosa, a temporary deficiency of DAO occurs. DAO activity was also normalized after the infection was cured.

In the context of the addition of a plurality of different substances which interfere with the activity, the DAO activity can be reduced exogenously. The substance preferably comprises an alcohol and acetaldehyde, a decomposition product thereof. Certain amine-rich foods and various drugs.

In all cases, the symptoms mentioned at the outset are more or less pronounced and, in most cases, cannot be easily classified. The rapid elucidation of the functional activity of the enzyme allows for rapid and simple treatment and establishment of a corresponding dietary regimen.

A common reason for the appearance of histamine intolerance is the sensitivity of the enzyme to a large number of chemicals. The substances are mostly present in a number of different drugs. The most important DAO inhibitors are acridine yellow, diazepam, N-methyl-N-formylhydrazine, B-aminopropionitrile, Dimaprit, O-methylhydroxylamine, Agmantine, ethanol (10%), propineb, methyldopa, furanilic acid, Phenamil, amiloride, guanabenz, phenelzine, aminoguanidine, guanfacine, pheneformin, amitriptyline, guanidine, Phenylrazine, amodiaquine, haloperidol, promethazine, anserine, benzylethylamine chloride, naphthylisopropamide, aziridinylalkylamine, hydroxychloroquine, B1 pyrimidine, hydroxylamine, quinine, cimetidine, semicarbazide, cadaverine, thiamine, carnosine, isoniazid, thioridazine, chlorothiazide, procarbazine, chlorpromazine, isoniazid, trimethoprim, thiamine, tryptophalmidine, tryptophalmine, clonidine, amitriptonidine, amitriptyline, clozapine, clozaprinine, clo, Metronidazole, tyramine, cyanide, nazlin (an alkaloid), a diamine (also called histamine), and Nt-methyl histamine.

In WO 02/43745 it is disclosed that DAO of plant origin is used systemically for the treatment of histamine mediated diseases. Application of DAO or enzymes (isolated directly from the plant) is generally problematic due to the often presence of allergens in the plant, especially in view of the fact that the pods disclosed in WO 02/43745 have a strong allergenic potential.

The object of the present invention was to provide compositions for the treatment and prevention of histamine-induced diseases and disorders which do not have the side effects of the products existing on the market (mainly comprising cortisone) and which eliminate the disadvantages of the prior art described above. It is a further object of the invention to increase the concentration of active diaminooxidase in the body, in particular in the intestine, of an individual in order thereby to promote or enable the breakdown of histamine which is added, in particular exogenously (for example by food).

The invention therefore relates to a pharmaceutical composition comprising diaminooxidase for the treatment of histamine-induced diseases, said composition being administered in the form of a transdermal, oral, peroral or sublingual administration, as a hydrogel, as a gastro-resistant pill, as drops, in particular eye drops and nasal drops, or as a tablet, a food supplement-composition or a nutraceutical and cosmetic composition comprising diaminooxidase. Diaminooxidases are present in the compositions of the invention predominantly in their active form, i.e. enzymes which, for example, do not have copper in their prosthetic groups and do not exhibit at least wild-type activity are unsuitable for the use according to the invention.

The pharmaceutical composition of the invention is administered in a form that enables administration in a manner selected from the group consisting of: transdermal, oral, peroral and sublingual. Epicutaneous application of DAO is particularly advantageous in the case of histamine-induced diseases or disorders on the skin surface or outermost layer of the skin, so that, for example, allergic reactions caused by skin contact with allergens can be successfully treated. Inflammation by histamine release in the case of various diseases such as urticaria, atopic eczema and the like can also be suppressed. By oral and peroral administration of the composition of the invention, DAO can be made to enter the gastrointestinal tract of an individual and there successfully inhibit or treat histamine-induced diseases by decomposing histamine. The scope of action of DAO is mainly limited to the intestinal tract, since high acid content in the stomach has a negative effect on DAO activity. Thus, oral or peroral administration of DAO requires protection from gastric acid until the intestinal tract is reached. Conversely, if DAO is administered sublingually, the enzyme is rapidly absorbed by the oral mucosa in the mouth and enters the blood vessels. It is thus possible to achieve rapid and simple transfer of DAO into the blood vessels without having to deliver the enzyme intravenously or intact via the stomach to the intestine, which delays the rapid onset of enzymatic action.

The pharmaceutical composition is provided in an administration form selected from the group consisting of: hydrogels, gastro-resistant pills, drops, especially eye and nose drops, tablets and capsules. By the present invention, DAO can be converted accordingly by processing methods known in the art into pharmaceutical administration forms. Thus, the pharmaceutical compositions comprising DAO also contain other inclusions, with which the enzyme is stabilized on the one hand and the enzyme is brought into the corresponding galenic form on the other hand. DAO can naturally also be administered in a single administration form together with other pharmaceutically active substances or separately, as long as the enzyme is not inhibited by one of these active substances to such an extent that the activity of the enzyme does not exert the desired effect.

Hydrogel-compositions of the invention comprising DAO at a shear rate of 41 seconds^-1It preferably has a viscosity of 0.5 to 5cp (centipoise), still more preferably 1 to 2cp, especially 1.1 to 1.6cp, wherein the measurement can be performed with a viscometer known in the art. It has been shown that, in particular in the viscosity range mentioned, DAO-containing hydrogel compositions have advantageous properties (e.g.local dispersibility, good handling). At a lower viscosity of 0.5cp, the hydrogel composition proved to be too thinSo thin to be flowable that the user does not handle the formulation well. As gelling agents, preference is given to polyacrylates (e.g. Carbopol), cellulose derivatives or modified celluloses, especially hydroxyethylcellulose (e.g. Natrosol), methylcellulose (Metylzellulose), hydroxypropylmethylcellulose and hydroxymethylcellulose, starch and modified starch, natural and synthetic rubbers, such as tragacanth, guar gum, carageenan, gelatin, sodium alginate, PVP, polyvinyl alcohol and mixtures thereof. Particular preference is given to using hydroxyethylcellulose (for example Natrosol) as gelling agent, with Natrosol Typ 250 HHR in particular having particularly good stability even at lower concentrations. The hydrogel composition preferably has a pH of 7.0 to 9.0, preferably 7.2 to 8.5, more preferably 7.5 to 8.0. The pH range is preferably adjusted with buffers, in particular Bis/Tris-buffers, wherein, in order to maintain the DAO enzyme activity and stability, further salts (for example 20 to 300mM, preferably 50 to 200mM, in particular 100mM NaCl) may also be added. If the total concentration of salts is less than 20mM, the DAO has poor stability, which results in loss of activity.

In addition, a preservative that mainly prevents the propagation of microorganisms in the composition may be added to the hydrogel composition of the present invention. It is important that the preservative is selected such that the antimicrobial substance does not inhibit DAO activity in such a way that its enzymatic activity is no longer sufficient, thereby achieving the object of the present invention (decomposition of histamine). Such substances can be identified by activity tests with DAO and preservatives. Chlorhexidine, parabens (p-hydroxy-benzoates, especially butyl, ethyl, methyl or propyl parabens), benzoates (e.g. sodium benzoate), sorbates (e.g. potassium sorbate), carbamates (e.g. iodopropynyl butyl carbamate) and combinations thereof (e.g. rokosol ═ a mixture of sodium benzoate, potassium sorbate and iodopropynyl butyl carbamate) are preferably used as preservatives. Particularly preferred are hydrogel compositions containing Natrosol as gelling agent and Rokosol and/or parabens as preservative. Obviously, it is also possible to add fragrances and aromatizers, for example, as used in the cosmetic industry, according to the invention.

Another aspect of the invention relates to food supplement-compositions or nutritional foods (DLM) adapted to act as gastro-resistant pills, drops or infusions. Oral administration of DAO by food, food supplements or nutritional foods presupposes that the enzyme is able to reach any part of the body where it is intended to exert its effect. Since histamine plays an important role in the intestinal tract, DAOs must be modified or configured such that they can pass through the stomach containing strong acids unaffected. The DAO is preferably processed here as a gastric resistant pill.

Since DAO is irreversibly destroyed at pH values of less than 3 (in the stomach, pH values between 2 and 4), in order for DAO to be transported through the stomach into the intestinal tract, DAO must be configured for corresponding administration (pharmaceutical or food supplement compositions or nutritional foods). The present invention demonstrates that capsules (e.g., gelatin capsules), and particularly gastric juice resistant pills, are particularly advantageous. The effect of DAO advantageously occurs at the latest 15, preferably at the latest 20, in particular at the latest 30 minutes after application.

According to the invention, this property of the pill is referred to as "gastric resistance", wherein the active substance (e.g. DAO) contained therein is protected under the action of gastric fluid or a solution having properties comparable to gastric fluid (e.g. acid) over a determined time interval of at least 10, preferably at least 20, more preferably at least 30, especially at least 60 minutes, such that the active substance suffers a loss of activity of at most 50%, preferably at most 40%, more preferably at most 30%, most preferably at most 20%, especially at most 10%.

Preferably the pellets of the invention release at least 60%, especially at least 80% of the DAO activity used to formulate the pellets into the intestine after 20 minutes, especially after 30 minutes.

A gastro-resistant pellet is a pellet surrounded by a gastro-resistant coating which dissolves at the pH values as present in the intestinal tract. That is, such a coating is therefore preferably dissolved at a pH value of at least 4 and at most 10. Eudragit, e.g. gastric resistant coatings based on anionic polymers of methacrylic acid and methacrylic acid esters, contains-COOH as functional group and dissolves in the pH range 5.5 to pH 7. Instead of Eudragit, shellac or acetylated starch (e.g. Amprac 01) may also be considered. Since gastric juice-resistant coatings known from the prior art have various properties (e.g. pH at which the coating dissolves, dissolution speed), it is also possible to use the coating materials in combination. Shellac, for example, exhibits good acid resistance, but dissolves very slowly in the intestinal tract. In contrast, Amprac01 dissolved rapidly in the intestinal lumen, but did not exhibit sufficient acid resistance. In order to compensate for the disadvantages of the materials, the two materials are mixed, for example, in a weight ratio of 60-95/40-5, preferably 70-90/30-10, shellac/Amprac 01. Another parameter that influences the release rate of the active substance is the layer thickness of the gastro-resistant pellets. The layer thickness, expressed in mass proportion, is here preferably from 5 to 30%, more preferably from 10 to 20%, of the total mass of the end product. The mean diameter of the pellets is preferably from 0.5 to 5mm, in particular from 0.7 to 2 mm. The advantage of such a size is that the pellets can pass through the stomach quickly.

The preparation of the pellets according to the invention, both for use in the pharmaceutical composition according to the invention and in the food supplement-composition or nutritional food according to the invention, is preferably carried out with an extruder, which requires a thermal stability of the contents of the composition, in particular of the active substance DAO, of up to 60 ℃ (strickera zrne mintworkling, spitnger Verlag 2003). The pellets may contain other pharmaceutical additives in addition to the gastro-resistant coating and the DAO. For example microcrystalline cellulose (e.g. Avicel) is used as filler and swelling agent. Cellulose is insoluble in water and has both crystalline and amorphous portions in its shape. This combination affects the plastic deformability, which means that irreversible shape changes occur with a sufficiently large force. This is a basic prerequisite for pelletizing in extruders and spheronizers. In the case of wet granulation, microcrystalline cellulose absorbs a large amount of water, and therefore can be formed into a mass that can be compressed and adhered well even without adding a binder. According to the invention, the amount of microcrystalline cellulose in the pill can be 5-70%, preferably 10-60%, more preferably 15-50%. Sucrose can be used as a binder and filler. Sucrose improves the solubility of the base material, thus facilitating the rapid release of the enzyme. According to the invention, sucrose may be added to the pellets in the range of 1 to 40%, preferably 5 to 35%, more preferably 10 to 30%. Hydroxypropyl cellulose (added preferably in an amount of 0.5 to 10%) may be similarly added as a binder for preventing dust. Furthermore, hydroxypropylcellulose also increases the strength of the pellets, thus again contributing to improved results. Corn starch may be added to the pellets of the present invention as a filler and disintegrant (preferably in an amount of 1 to 30%). Starch, as a water-insoluble substance, can absorb large amounts of water and is therefore an ideal disintegrant. Cross-linked carboxymethylcellulose (Na-CMC; Acdisol) is a pure disintegrant, preferably used in an amount of 1% to 5%. Too high an Acdisol content can lead to premature disintegration of the pellets, even when rounded off, and hence a kontrakproduktiv. Crosslinked polyvinylpyrrolidone (crospovidone), a laterally crosslinked PVP, is also water insoluble and is also useful as a disintegrant. Due to its polymeric nature, better rounding is promoted when making pellets (preferred addition levels may be 0.5 to 10%). Polyvinylpyrrolidone is a water-soluble additive used as a binder. The combination of these different fillers, disintegrants and binders allows the DAO molecules to be dispersively distributed in the pellet and ensures rapid bioavailability.

A barrier layer formed of glycerol and/or talc may be provided between the gastro-resistant coating and the active substance-containing pellets. Glycerol is used as a humectant to prevent dehydration and thus inactivation of enzymes.

Instead of a pill, DAO in a capsule can also be delivered through the stomach into the intestine. Suitable capsules are here, for example, gelatin capsules or starch capsules. The capsules may also be included in the pills of the present invention.

Another aspect of the invention relates to a cosmetic composition comprising a diaminooxidase in a form for cosmetic administration, especially as a hydrogel, ointment, spray or drops. At higher histamine release or upon contact with allergenic substances (e.g. upon contact with an allergen or neurodermatitis), a physical reaction occurs in the visible part of the body, which can be inhibited by the application of DAO in a cosmetic composition. Cosmetic compositions containing DAO may also have other inclusions known in the art, which are used in the preparation of cosmetic products. The cosmetic composition containing the hydrogel has substantially the same properties as or contains substantially the same contents as the hydrogel of the pharmaceutical composition of the present invention.

The diaminooxidase used in the compositions of the present invention is preferably not of vegetable origin.

The use of DAO that is not of vegetal origin in pharmaceutical and cosmetic compositions, as well as food supplements and nutritional foods, has the advantage that the presence of allergens in plants does not negatively affect the administration of DAO, since allergens mainly require an endogenous histamine release. It is known that, in particular, vegetable substances are responsible for histamine-induced diseases. Complete removal of allergenic inclusions from DAO-preparations of vegetable origin can only be achieved with high production costs, whereas the DAO of the invention with non-vegetable origin is completely free of such vegetable allergens.

According to the invention, "non-vegetal origin" includes all DAOs obtained not from plants but from the animal body or other non-vegetal bodies. Furthermore, according to the invention, this definition includes all DAOs produced recombinantly in cell culture (animals, bacteria, yeasts, etc.) or in any kind of non-vegetal organism, wherein the DNA from the plant and/or animal organism is isolated by methods known in the art and cloned and expressed in an expression system for the recombinant production of DAOs.

Preferably all compositions disclosed in the present invention contain animal derived diaminooxidases. By using animal DAOs, enzymes can be supplied to the human or animal body such that the enzymes produced by these individuals themselves are very close in glycosidation, activity and specificity to the DAOs produced by these individuals themselves. In addition, botanical allergens can be completely excluded from the preparation of DAO.

The diaminooxidase is preferably obtained from pig kidney. Porcine kidneys are particularly characterized by their high DAO content. The enzyme can be easily isolated from porcine kidney by methods known in the art.

According to another preferred embodiment, the composition of the invention contains a diaminooxidase of recombinant origin. By recombinantly producing DAO, a large amount of enzyme can be produced and the enzyme is purified in high yield.

Preferably the recombinant diaminooxidase is expressed in prokaryotes, preferably in bacteria, or in eukaryotes, preferably in animal or yeast cell culture, and isolated from the expression system described above. The DAO produced by the expression system is purified and either expressed in the cell or isolated from the cell during expression by methods known in the art. In this case, it is also possible to provide the DAO with a peptide (e.g.His-Tag), polypeptide or protein sequence (e.g.GST-Tag), so that the purification can be simplified. Recombinant DAO can also be modified by gene technology methods such that the enzymatic activity of the DAO exceeds that of wild-type DAO.

Another aspect of the invention relates to the use of a diaminooxidase of the invention for the preparation of a medicament for the treatment of histamine-induced disorders. DAO is known to be responsible for the breakdown of histamine in humans and animals. Because histamine-induced diseases are caused by an excess of histamine due to a deficiency of diaminooxidase, by inhibition of DAO or by an excess of histamine, which is usually due to food, but also due to other external factors such as exposure to allergens, administration of DAO according to the invention is suitable for the treatment of these diseases or disorders.

Another aspect of the invention relates to the use of the diaminooxidase of the invention for the preparation of a medicament for the treatment of urticaria, in particular chronic and acute urticaria. In the case of urticaria, the release of histamine expands the venules and increases the throughput of capillaries with resulting edema. By administering DAO to the relevant skin area, histamine in the relevant site can be decomposed, thereby inhibiting the itching irritation of urticaria.

Another aspect of the invention relates to the use of a diaminooxidase of the invention for the preparation of a medicament for the treatment of contact allergies. Contact allergy is caused by substances (allergens) which initiate an allergic reaction by penetrating the skin. In order to decompose histamine released by such contact and thereby inhibit or alleviate histamine-induced conditions, DAO is administered.

According to another aspect of the invention, the diaminooxidase of the invention is used for the preparation of a medicament for the treatment of atopic dermatitis. Atopic dermatitis, also known under the name neurodermatitis, is a common skin disease with strong itching stimuli, mostly during the growth phase of children and adolescents. The cause of the inflammation is an excessive release of histamine at the relevant skin site. DAO may also in this case contribute to reducing histamine in the area, thus alleviating or preventing symptoms of atopic dermatitis.

According to another aspect, the invention relates to the use of a diaminooxidase according to the invention for the preparation of a medicament for the treatment of scombroid intoxication. Scomber scombrus poisoning is histamine poisoning caused by eating Scomber scombrus such as tuna. When the cold chain was interrupted or cooking was delayed, Scombriden formed so-called mackerel toxins, which resulted in histamine enrichment. The consequences of histamine poisoning are, inter alia, fever, nausea, vomiting, abdominal pain and urticaria. DAO may be used as an antidote in this case.

Another aspect of the invention relates to the use of the diaminooxidase of the invention for the preparation of a medicament or a food supplement or a nutritional food for the removal or reduction of histamine in the gastrointestinal tract.

The reduction of histamine from the intestine is of particular interest when increasing the addition of histamine to the body of an individual (exogenous addition, e.g. by food) or, for example, when the activity of DAO in the intestine is partially or completely inhibited or absent. Thus, the medicament or food supplement or nutraceutical of the invention is used to add enzymatically active DAO, which aids in the breakdown of histamine in the intestinal tract.

Another aspect of the invention relates to the use of a diaminooxidase for the preparation of a medicament for the removal of histamine from the bronchial system.

Absorption of histamine-releasing substances, such as pollen, can lead to strong allergic reactions in the bronchial system or in the lungs. In order to decompose the released histamine, the DAO of the invention may be introduced into the bronchial system or lungs, for example, by inhalation spray or the like.

The invention is further illustrated by the following examples, but is not limited thereto.

Example (b):

example 1

DAO is actively stabilized in cellulose-based pellets and the pellets are coated with a layer resistant to gastric juices. Dissolution testing indicated that more than 70% of the activity was released into the environment within the first hour.

Example 2

In addition, the enzyme is stabilized in the hydrogel. Storage at room temperature and 37 ℃ showed no decrease in activity over 4 months. Blisters and inflammation on the skin after stimulation with histamine disappeared after a few minutes after application of the hydrogel.

Example 3

Stabilized DAO in hydrogels was tested in a total of 13 people. With the help of questionnaires, the suffering from contact allergy (n ═ 5), neurodermatitis/skin diseases (n ═ 6) and insect bites (n ═ 2) are distinguished. Positive effects were tested for all contact allergic patients and neurodermatitis patients, and for insect bites, 50% of subjects could be reported as positive effects.

Disease of the disease	Positive Effect/number of subjects
		Neurodermatitis/skin diseases	6/6
Contact allergy	5/5
		Insect bites and stings	1/2

The effect of the hydrogel appeared within the first 10 minutes, which was maintained on average longer than one hour.

	0-10 min	10-30 minutes	30-60 minutes	＞1h	＞3h
						Start of action	11	1
Sustained action		3	1	2	6

Upon repeated use, no subjects claim an unpleasant effect.

Two patients with neurodermatitis indicated that the hydrogel of the invention was more effective than the cortisone ointment currently used.

Example 4

Gastric resistant pellets were prepared with 3% DAO (demonstrated an initial activity of 80000E/ml), 40% microcrystalline cellulose, 20% sucrose, 22% other binders, fillers and disintegrants and with a 15% gastric resistant coating. It was observed by activity experiments that DAO was released from the pellets over a period of 180 minutes. It is demonstrated here that the DAO activity of the solution in which the pellets are dissolved after 10 minutes is 40% of the amount of DAO initially used, 70% after 30 minutes, 80% after 60 minutes and 100% after 180 minutes. The activity measurement of DA0 was carried out as described for AT 411688, but it is entirely possible to measure the enzyme activity using other known methods.

The literature:

Bachrach U.，in：B.Mondovi(Ed)(1985)，Structure and Functionsof Amine Oxidase，CRC Press，Boca Raton，pp 5-20xBartholomew M.J.et al(1990)，Cancer 66：1539-1543

James S.M.，Palcic M.M.，Scaman C.H.，Smith A.J.，Brown D.E.，Dooley D.M.，Mure M.，Klinman J.P.(1992)，Biochemistry31：12147-12154

James S.M.，Mu D.，Wemmer D.，Smith A.J.，Kaur S.，Maltby D.，Burlingame A.L.，Klinman J.P.(1990)，Science 248：981-987

Keskinege A.，Elgun S.，Yilmaz E.(2001)，Biochim Biophys Acta25：76

Kitanaka J.，Kitanaka N.，Tsujimura T.，Terada N.，Takemura M.(2002)，European Journal of Pharmacology 437：179-185

Kluetz M.D.，Schmidt P.G.(1977)，Biochem.Biophys.Res.Comm.76：40-45

Küfner M.A.，Ulrich P.，Raithel M.，Schwelberger H.G.(2001)，Inflamm.res.50，Supplement 2，96-97

Kusche J.，Menningen R.，Leisten L.，Krakamp B.(1988)，Adv.Exp.Med.Biol.250：745-52

Kusche J.，Bieganski T.，Hesterberg R.，Stahlknecht C.D.，Feuss-ner K.D.，Stahlenberg I.Lorenz W.(1980)，Agents Actions10：110-3

Mennigen R，Bieganski T，Elbers A，Kusche J(1989)，J ChromatogrB Biomed Sci Appl 27：221

Mizuguchi H.，Imamura I.，Takemura M.，Fukui H.(1994)，J.Bio-chem 116：631-635

Mu D.，James S.M.，Smith A.J.，Brown D.E.，Dooley D.M.，KlinmanJ.P.(1992)，J.Biol.Chem.267：7979-7982

Nilsson B.O.，Kockum I.，Rosengren E.(1996)，Inflammation Res.45：513-518

Rinaldi A.，Vecchini P.，Floris G.(1982)，Prep.Biochem.12：11-28

Shah M.A.，Ali R.(1988)，Biochem.J.253：103-107

Taylor S.L.(1986)，Crit.Rev.Toxicol.17：91-128

Claims (18)

1. Pharmaceutical composition comprising diaminooxidase for the treatment of histamine-induced diseases, characterized in that the composition is administered in the form of a transdermal, oral, peroral or sublingual administration, as a hydrogel, as a gastro-resistant pill, as drops, especially eye drops and nasal drops, or as a tablet.

2. Food supplement-composition comprising diamino oxidase.

3. A nutritional food comprising a diamino oxidase.

4. Composition according to claim 2 or 3, characterized in that it is in the form of pellets, drops or infusions resistant to gastric juices.

5. Cosmetic compositions comprising diaminooxidase.

6. Composition according to claim 5, characterized in that it is in the form of a cosmetic administration, in particular as a hydrogel, ointment, spray or drops.

7. Composition according to one of claims 1 to 6, characterized in that the diaminooxidase is of non-vegetal origin.

8. Composition according to claim 7, characterized in that it comprises a diaminooxidase of animal origin.

9. The composition of claim 8, wherein the diaminooxidase is from porcine kidney.

10. Composition according to one of claims 1 to 9, characterized in that it comprises a diaminooxidase of recombinant origin.

11. Composition according to claim 10, characterized in that the composition comprises a recombinant diaminooxidase obtained from prokaryotes, preferably from bacteria, or from eukaryotes, preferably from cell cultures of animals or yeasts.

12. Use of a diaminooxidase for the preparation of a medicament for the treatment of histamine-induced disorders.

13. Use of diaminooxidase for the preparation of a medicament for the treatment of urticaria, especially chronic and acute urticaria.

14. Use of a diaminooxidase for the preparation of a medicament for the treatment of contact allergies.

15. Use of a diaminooxidase for the preparation of a medicament for the treatment of atopic dermatitis.

16. Use of diamino oxidase in preparing medicine for treating scombrus poisoning.

17. Use of a diaminooxidase for the preparation of a medicament for removing histamine from the gastrointestinal tract.

18. Use of a diaminooxidase for the preparation of a medicament for the removal of histamine from the bronchial system.