ES2344915T3 - PROCEDURE FOR PREVENTIVE ALLERGY TREATMENT BY MUCOSAL ADMINISTRATION OF AN ALLERGY VACCINE. - Google Patents

Abstract

An allergen for use in allergy preventive treatment by administering an allergy vaccine comprising the allergen to a mucosal surface of a subject, a) in which the subject to be treated is sensitized such as to present a specific IgE response for the allergen, b) in which the subject to be treated is free of allergy clinical symptoms associated with the allergen, and c) in which preventive treatment is directed to avoid or reduce subsequent clinical allergy symptoms associated with the allergen.

Description

Preventive treatment procedure of Allergy by mucosal administration of an allergy vaccine.

Technical field

The present invention relates to a allergen allergy preventive treatment procedure in a subject comprising administering an allergy vaccine to a mucosal surface in the subject.

Background of the invention

Allergy is a major health problem in countries where the western lifestyle is adapted. Besides, the prevalence of allergic disease is increasing in these countries Although allergy in general cannot be considered a life-threatening disease, asthma causes an annual significant number of deaths. An exceptional prevalence of approximately 30% in adolescents transmit a loss substantial in quality of life, days of work and money, and justifies a classification among the main health problems  in the western world

Allergy is a complex disease. Many factors contribute to the sensitization event. Among these is the susceptibility of the individual defined by an even Insufficiently understood interaction between several genes. Other important factor is an allergen exposure above certain thresholds Some environmental factors may be important in the sensitization process including pollution, infections children, parasite infections, microorganisms intestinal, etc. Once an individual is sensitized and the allergic immune response is established, the presence of only tiny amounts of allergen effectively translates into symptom.

The natural course of allergic disease is usually accompanied by aggravation at two levels. Firstly, a progression of symptoms and severity of the disease, as well as disease progression, for example from rhinitis Pollinic to asthma.

Second, the spread in allergens that cause discomfort most often results allergic multreactivity. Chronic inflammation leads to a general weakening of mucosal defense mechanisms giving as a result nonspecific irritation and eventually destruction of the mucosal tissue. Young children can reach fundamentally sensitize against food, that is milk, resulting in eczema or gastrointestinal disorders; without However, most often they overcome these symptoms spontaneously. These young children are at risk of develop inhalation allergy later in their lives.

The most important allergen sources are found among the most prevalent particles of a certain size In the air we breathe These sources are remarkably universal and include grasses of grasses and faecal particles of house dust mites, which are jointly responsible for Approximately 50% of all allergies. Of global importance it is also animal dander, for example dog dander and cats, other pollens, such as Artemis pollens, and micro-fungi, such as Alternaria. On a base regional still other pollens can dominate, such as pollen from birch in northern and central Europe, ragweed pollen in the Eastern and central United States and cedar pollen from Japan in Japan. Insects, that is, bee and wasp venoms, and food accounts for approximately 2% of all allergies

The allergy, that is the hypersensitivity of type I, is caused by an inappropriate immune reaction to foreign non-pathogenic substances. Clinical manifestations Important allergies include asthma, pollen rhinitis, eczema and gastrointestinal disorders The allergic reaction is rapid and reaches its peak in 20 minutes after contact with the allergen that causes discomfort. In addition, the allergic reaction is specific in the sense that a particular individual is sensitized to a particular allergen (s), while the individual does not necessarily show a reaction allergic to other substances known to cause disease allergic The allergic phenotype is characterized by a pronounced inflammation of the mucosa of the target organ and by the presence of allergen specific antibody of the IgE class in the circulation and on the surface of primed cells and basophils

An allergic attack is initiated by the reaction of foreign allergen with allergen-specific IgE antibodies, when antibodies are bound to specific IgE receptors High affinity on the surface of mast cells and basophils. Prime cells and basophils contain mediators preformed, ie histamine, tryptase, and other substances, which they are released after crosslinking of two or more IgE antibodies. The IgE antibodies are crosslinked by the simultaneous binding of a allergen molecule. It follows therefore that a foreign substance that has only one non-allergen binding epitope initiates an allergic reaction IgE crosslinking bound to receptor on the surface of the primed cells also leads to release signaling molecules responsible for the attraction of eosinophils, allergen-specific T lymphocytes, and other types of cells at the site of the allergic response. These cells in interaction with allergen, IgE and effector genes, lead to renewed lightning of symptoms that occur 12-24 hours after encounter with allergens (late phase reaction).

The management of allergic disease includes diagnosis and treatment including prophylactic treatments. The Allergy diagnosis is affected by the manifestation of IgE allergen specific and by identifying the source of allergens In many cases a careful history can be sufficient for allergy diagnosis and for identification of the allergen source material that causes the discomfort. The most Frequently, however, the diagnosis is supported by objective measures, such as simple cutaneous puncture test, blood test, or provocation test.

The therapeutic options are given in three main categories The first opportunity is the avoidance of Allergen or exposure reduction. While avoidance of the allergen is obvious, for example in the case of allergens food, it can be difficult or expensive, as for allergens of homemade dust mites, or it may be impossible, as for pollen allergens The second and most widely used option therapeutic is the prescription of classic symptomatic drugs as antihistamines and steroids. Symptomatic drugs are safe and efficient, however, they do not alter the natural cause of the disease, nor do they control the spread of the disease. The third therapeutic alternative is specific allergy vaccination which in most cases reduces or relieves symptoms allergies caused by the allergen in question.

Conventional Specific Allergy Vaccination It is a causal treatment for allergic disease. She interferes with basic immune mechanisms resulting in persistent improvement of the immune status of patients. Thus, the specific allergy vaccination protective effect extends more beyond the treatment period in contrast to the treatment of symptomatic drug Some patients receiving treatment they heal, and in addition, most patients experience a relief in the severity of the disease and symptoms experienced, or at least one detention in the worsening of the disease. Thus, vaccination of specific allergy has effects preventive measures that reduce the risk of pollinic rhinitis evolve to asthma, and that reduce the risk of developing new sensibilities.

The immune mechanism that underlies the Successful allergy vaccination is not known in detail. An answer  specific immune system, such as the production of antibodies against a particular pathogen, is known as an immune response adaptive This answer can be distinguished from the answer Innate immune, which is a nonspecific reaction to pathogens. A allergy vaccine is linked to targeting the immune response adaptive, which includes cells and molecules with specificity antigenic, such as T lymphocytes and the B lymphocytes that produce antibodies B lymphocytes cannot mature in cells that produce antibodies without the help of T lymphocytes from the corresponding specificity. T lymphocytes that participate in the Stimulation of allergic immune responses are mainly of type Th2. It has been proposed that the establishment of a new balance between Th1 and Th2 lymphocytes is beneficial and fundamental for the immunological mechanism of specific allergy vaccination. If this is caused by a reduction in Th2 lymphocytes, a change of Th2 to Th1 lymphocytes, or upward regulation of Th1 lymphocytes It is controversial. Recently, it has been proposed that lymphocytes T regulators are important for the vaccination mechanism of allergy. According to this model the regulatory T lymphocytes, that is the Th3 or Tr1 lymphocytes, it would regulate both the Th1 lymphocytes such as Th2 of antigen specificity correspondent. Despite these ambiguities it is generally believed that an active vaccine must have the ability to stimulate allergen-specific T lymphocytes, preferably those TH1 lymphocytes.

The specific allergy vaccination is not, to in spite of its virtues, of extended use, fundamentally by two reasons. One reason is the inconvenience associated with the program of traditional vaccination comprising repeated vaccinations among other things injections for several months. The other reason is, more importantly, the risk of side reactions allergic Ordinary vaccinations against infectious agents are carried out efficiently using a single immunization of high dose or a few high dose immunizations. This strategy, however, cannot be used for allergy vaccination since an immune response is already in progress.

Conventional Specific Allergy Vaccination it is therefore carried out using multiple immunizations subcutaneous applied for a prolonged period of time. He course is divided into two phases, the dose increase phase and the maintenance phase In the dose increase phase they are applied increasing doses, typically over a period of 16 weeks, Starting with tiny doses. When the dose of recommended maintenance, this dose is applied during the phase of maintenance, typically with injections every six weeks. After each injection the patient must remain in medical care for 30 minutes due to the risk of side reactions anaphylactic, which in principle although extremely rare could Be life threatening. In addition, the clinic should be equipped to maintain emergency treatment. There's no doubt about that a vaccine based on a different route of administration eliminate or reduce the risk of side reactions allergies inherent in the vaccine based on the current subcutaneous route as well as facilitate more widespread use, possibly allowing even self-vaccination at home.

Attempts to improve vaccines for vaccination Specific allergy have been carried out for more than 30 years and include varied approaches. Several approaches have addressed the allergen itself by modifying the reactivity of the IgE.

Fanta et al. (Int. Arch. Allergy Immunol., 1999, 120: 218-224) refers to a study of changes immune-induced SLIT in a group of patients allergic to grass pollen selected according to the criteria of have clinical symptoms (rhinitis and / or seasonal bronchial asthma) during the grass pollen season, a puncture test simple cutaneous positive to grass pollen extracts, and IgE specific for grass pollen as determined by RAST-CAP. The SLIT was carried out by administration Sublingual drops of allergen extract.

Holt et al. ("Suppression of IgE responses following inhalation of antigen ", Immunology Today, volume 8, No. 1, 1987) mentions the fact that as a response to exposure to allergen inhaled or instilled oronasally, is induced tolerance in the upper respiratory tract corresponding to that induced in the gastrointestinal tract by antigens food

Holt et al. ("Sublingual allergen administration. I. Selective suppression of IgE production in rats by high allergen doses ", Clinical Allergy, 1988, Volume 18, pages 229-234) refers to the administration sublingual of an allergen (ovalbumin) to rats that have not previously treated for seven consecutive days followed by a ovalbumin challenge test five days after Last sublingual dose. The results showed a suppression selective IgE specific for ovalbumin. It is speculated that the treatment mechanism involves cell stimulation specific allergen suppressors. It is further mentioned that The proposed treatment involves animals that have not been treated above and should be distinguished from a procedure of sublingual desensitization.

WO 95/17208 discloses a procedure of allergic disease prevention that includes administering to a previously not sensitized subject an effective allergen dose to induce the establishment of a stable population of memory lymphocytes similar to helper T lymphocytes 1 Allergen-specific capable of inhibiting lymphocyte activity memory similar to T helper lymphocytes 2. The subject to Treat is preferably between 3 months and 7 years. How allergens are mentioned for example homemade dust mites, pollen of grasses and tree pollen. Allergen Administration it can be carried out orally, intranasally, oronasally, rectal, intradermal, intramuscular or subcutaneous.

The homepage www.immunotolerance.org (October 11, 2004) describes for example a planned clinical study of preventive treatment of children without sensitization to inhalants, in which sublingual drops containing any allergens (homemade dust mites, flelen pollen from the meadows and cat) to children, and in which children are monitored for the development of allergy for three years. Children recruited for the study have a history of atopic dermatitis or food allergy and their biological mother or biological father or brother or sister has a history of atopy.

US 6,333,038 discloses regulation of development of normal anti-allergenic immunity administering allergen to children, who have not been sensitized.

The objective of the present invention is provide an improved preventive treatment procedure for individuals, particularly children.

Summary of the invention

This objective is obtained with the present invention, which refers to an allergen for use in treatment allergy prevention by administering an allergy vaccine that contains the allergen to a mucosal surface of the subject,

to)

at that the subject to be treated is sensitized as to present an allergen-specific IgE response,

b)

at that the subject to be treated is free of allergy clinical symptoms associated with the allergen, and

C)

at that preventive treatment is aimed at avoiding or reducing symptoms subsequent allergy clinics associated with the allergen

The invention is based on the novel finding of that it is possible to avoid allergy symptoms for an allergen from of development in an individual whose immune system has been exposed to the allergen, but in which the immune response has not progressed even to a state that involves clinical symptoms, such such as rhinitis, conjunctivitis, rhinorrhea, nasal obstruction, sinusitis, sneezing, atopic dermatitis, itching, watery eyes, runny nose, wheezing and skin irritation. It has been believed to date that in order to achieve preventive treatment Effective of an allergy the individual to be treated should be not sensitized In addition, it has been believed to date that the mechanism involved in the prevention of an allergy is tolerance induction oral that corresponds to that induced in the tract gastrointestinal by dietary agents. However, I agree with the present invention it has been surprisingly shown that it is possible to achieve a preventive effect by treating individuals, who already They are sensitized and do not show any clinical symptoms. It is believed that the preventive effect obtained takes place through a mechanism, which is partially or totally similar to the mechanism of desensitization that takes place in allergy vaccination specific (immunotherapy).

It is additionally believed that the treatment preventive is more effective when carried out so soon after  of sensitization as possible before the response of the immune system begin to change additionally to a response of allergic Th2 lymphocytes. In other words it is in general advantageous to treat children as young as possible once you have been exposed to an allergen In addition, it is believed that due to the effectiveness of such early preventive treatment, the treatment It can be done with smaller doses, less administrations and / or a shorter treatment period compared to vaccination of specific allergy of adults with developed clinical symptoms. Due to the smoothness of the preventive treatment protocol, it is suitable for use in general vaccination programs of all children or large groups of selected children.

An additional advantage of the invention is that sensitized people are the most likely people they develop allergy, and thus sensitized people constitute the most relevant group of people to undergo treatment preventive.

The invention also relates to the use of a allergen for the manufacture of an allergy vaccine that understands the allergen as an active substance for treatment allergy prevention by administering the allergy vaccine to a mucosal surface of a subject,

to)

at that the subject to be treated is sensitized as to present an allergen-specific IgE response,

b)

at that the subject to be treated is free of allergy clinical symptoms associated with the allergen, and

C)

at that preventive treatment is aimed at avoiding or reducing symptoms subsequent allergy clinics associated with the allergen

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Short description of the figures

Fig. 1A shows clinical data (number of sneezing) in sensitized mice tested provocation intranasally either with Phl p of 5000 SQ or with Tampon.

Fig. 1B shows the level of serum IgE in sensitized mice undergoing challenge testing intranasally well with Phl p of 5000 SQ or with Buffer.

Figure 1C shows the level of BAL IgE in sensitized mice undergoing challenge testing intranasally well with Phl p of 5000 SQ or with Buffer.

Figure 1D shows the level of NAL IgE in sensitized mice undergoing challenge testing intranasally well with Phl p of 5000 SQ or with Buffer.

Figure 1E shows the level of eosinophils of NAL in sensitized mice undergoing challenge testing intranasally well with Phl p of 5000 SQ or with Buffer.

Figure 2A shows clinical data (number of sneezing) in sensitized mice well treated SLIT or buffer treatment and tested for provocation intranasally with Phl p.

Figure 2B shows hypersensitivity of the pathways Respiratory in response to methacholine challenge test (as measured by Penh values) in sensitized mice undergoing either SLIT treatment or buffer treatment and underwent provocation test intranasally with Phl p.

Figure 3A shows the level of serum IgE in sensitized mice subjected to either SLIT treatment or to buffer treatment and tested for provocation intranasally with Phl p.

Figure 3B shows the level of serum IgG1 in sensitized mice undergoing either SLIT treatment or either to buffer treatment and undergoing provocation test intranasally with Phl p.

Figure 4A shows the level of BAL IgE in sensitized mice subjected to either SLIT treatment or to buffer treatment and tested for provocation intranasally with Phl p.

Figure 4B shows the level of NAL IgE in sensitized mice subjected to either SLIT treatment or to buffer treatment and tested for provocation intranasally with Phl p.

Figure 4C shows the level of BAL IgA in sensitized mice subjected to either SLIT treatment or to buffer treatment and tested for provocation intranasally with Phl p.

Figure 4D shows the level of NAL IgE in sensitized mice subjected to either SLIT treatment or to buffer treatment and tested for provocation intranasally with Phl p.

Figure 5A shows the peroxidase level Eosinophilic in BAL in sensitized mice well subjected to SLIT treatment or buffer treatment and subjected to provocation test intranasally with Phl p.

Figure 5B shows the peroxidase level Eosinophilic NAL in sensitized mice well subjected to SLIT treatment or buffer treatment and subjected to provocation test intranasally with Phl p.

Figure 6A shows the proliferation of T lymphocytes in the spleen in sensitized mice well subjected to SLIT treatment or buffer treatment and subjected to provocation test intranasally with Phl p.

Figure 6B shows the proliferation of T lymphocytes in lymph nodes (LN) in mice sensitized undergoing either SLIT treatment or Buffer treatment and provocation test intranasally with Phl p.

Detailed description of the invention Allergen

The allergen according to the present invention can be any protein that occurs in nature that has been statement that induces allergic reactions, for example mediated by IgE after repeated exposure to an individual.

Examples of allergens that occur in the nature include pollen allergens (pollen allergens from trees, herbaceous plants, weeds and grasses), insect allergens (allergens of inhalants, saliva and poisons, for example allergens of mites, cockroaches and small mosquitoes, allergens of hymenoptera venoms), animal hair and animal dander allergens (for example dog, cat, horse, rat, mouse etc.), and food allergens. Important pollen allergens of trees, grasses and plants herbaceous are such that they originate from orders Taxonomic of Fagales, Oleales, Pinales and Platanáceas including among others birch (Betula), alder (Agnus), hazelnut (Corylus), carpe (Carpinus) and olive (Olea), cedar (Cryptomeria and Juniperus), banana (Platanus), the order of the Poales including among others grasses of the genera Lolium, Phleum, Poa, Cynodon, Dactylis, Holcus, Phalaris, Secale and Sorghum, the orders of the Asterales and Urticals including among other herbaceous plants of the Genres Ambrosia, Artemisia and Parietaria. Other allergens of Important inhalation are those of homemade dust mites of the Genus Dermatophagoides and Euroglyphus, warehouse mite for example Lepidoglyphys, Glycyphagus and Tyrophagus; those of cockroaches, small mosquitoes and fleas for example Blatella, Periplaneta, Chironomus and Ctenoceppalides, and those of mammals such as cat, dog and horse, poison allergens including those that originate from biting or biting insects such as those of the taxonomic order of the Hymenoptera, including bees (Apidae superfamily), wasps (Vespidea superfamily) and ants (Formicoidae superfamily). Inhalation allergens important fungi are among others those that originate from from the genera Alternaria and Cladosporium.

In a particular embodiment of the invention, the allergen is Bet v 1, Aln g 1, Cor a 1 and Carb b 1, Que a 1, Cry j 1,
Cry j 2, Cup a 1, Cup s 1, Jun a 1, Jun a 2, Jun a 3, Ole e 1, Lig v 1, Pla l 1, Pla a 2, Amb a 1, Amb a 2, Amb t 5,
Art v 1, Art v 2, Par j 1, Par j 2, Par j 3, Sal k 1, Ave e 1, Cyn d 1, Cyn d 7, Dac g 1, Fes p 1, Hol l 1, Lol p 1 and 5,
Pha a 1, Pas n 1, Phl p 1, Phl p 5, Phl p 6, Poa p 1, Poa p 5, Sec c 1, Sec c 5, Sor h 1, Der f 1, Der f 2, Der p 1, Der p 2,
Der p 7, Der m 1, Eur m 2, Gly d 1, Lep d 2, Blo t 1, Tyr p 2, Bla g 1, Bla g 2, Per a 1, Fel d 1, Can f 1, Can f 2, Bos d 2,
Equ c 1, Equ c 2, Equ c 3, Mus m 1, Rat n 1, Apis m 1, Api m 2, See v 1, See v 2, See v 5, Dol m 1, Dil m 2, Dol m 5,
Pol a 1, Pol a 2, Pol a 5, Sol i 1, Sol i 2, Sol i 3 and Sol i 4, Alt a 1, Cla h 1, Asp f 1, Bos d 4, Mal d 1, Gly m 1, Gly m 2,
Gly m 3, Ara h 1, Ara h 2, Ara h 3, Ara h 4, Ara h 5 or hybrids with their exchanged fragments of Molecular Breeding or any of these.

In a preferred embodiment of the invention the allergen is selected from the group consisting of an allergen of tree pollen, an allergen of grass pollen, an allergen of dust mites, an allergen from herbaceous plants and an allergen animal. Preferably, the allergen is selected from the group constituted by an allergen of grass pollen, an allergen of dust mites, an ambrosia allergen, a cedar pollen, a Cat allergen and a birch allergen.

In yet another embodiment of the invention the formulation comprises at least two different types of allergens that originate from the same allergenic source or originate from different allergenic sources for example allergens of group 1 of grasses and allergens of group 5 of grasses or allergens of group 1 of mites and allergens of group 2 of mites from different species of mites and grasses respectively, weed antigens such as ragweed allergens common and of giant ambrosia, allergens of different fungi like Alternaria and Cladosporium, tree allergens as allergens of birch, hazelnut, carpe, oak and alder, food allergens like Peanut, soy and milk allergens.

The allergen incorporated into the formulation it may be in the form of an extract, a purified allergen, a modified allergen, a recombinant allergen or a mutant of a recombinant allergen An allergenic extract may contain naturally form one or more isoforms of the same allergen, while a recombinant allergen typically represents only one isoform of an allergen In a preferred embodiment the allergen is in Extract form. In a preferred embodiment the allergen is a low IgE binding mutant or a low IgE binding mutant recombinant

Allergens may be present in equimolar amounts or the proportion of allergens present It can preferably vary up to 1:20.

In a preferred embodiment of the invention the Low IgE binding allergen is an allergen according to WO 99/47680, WO 02/40676 or WO 03/096869 A2.

Preventive treatment

Specific allergy vaccination (SAV), formerly known as specific immunotherapy or Hyposensitization, has been used for the treatment of disease Type 1 IgE-mediated allergy since the beginning of this century.

The general benefits obtained by SAV are: a) reduction of allergic symptoms and consumption of medicines, b) increased tolerance to allergens in the eyes, nose and lungs and c) reduced skin reactivity (phase reactions early and late phase).

The basic mechanism behind the improvement obtained by SAV it is unknown, but you can take out a series of common characteristics from the numerous studies of SAV carried out in recent decades: 1) the amount of total IgE is unchanged during the treatment period, 2) the amount of Allergen-specific IgE increases transiently during the dose increase, then falls back to the initial level (pretreatment), 3) epitope specificity and affinity of IgE remain unchanged, 4) allergen specific IgG, in particular IgG4, rises sharply during SAV, 5) starts apparently a new response from Th0 / 1 / Reg and 6) the response of Th2 seems unchanged. There is no correlation between the induced effect by SAV and the appearance of specific IgG.

SAV induces a new immune response that matures during the treatment period (T lymphocytes are recruited Th0 / 1, an allergen specific IgX is initiated (X can be A1, A2, G1, G2, G3, G4, M or D)). As the affinity (or quantity / affinity) of the new antibody response, IgX, has matured, IgX can compete efficiently with IgE for the allergen (s), inhibiting allergic response based on "normal" Th2 characterized by cross-linking of receptor-bound IgE in the surface of primed cells and basophils. So, the Clinical symptoms will be gradually reduced.

It is believed that preventive treatment led to implemented in the present invention works at least partially by means of the same mechanisms as revealed above to SAV.

The mucous membrane to which the allergy vaccine can be any mucous membrane adequate, and administration includes oral administration (through of the mucous membrane of the digestive system), nasal, vaginal, sublingual, ocular, rectal, urinary, intramammary, pulmonary, otic (i.e. through the ear) and buccal, preferably oral or sublingual administration (oromucosal administration). The Allergy vaccine can be in the form of a sprayer, an aerosol, a mixture, a suspension, a dispersion, a emulsion, a gel, a paste, a syrup, a cream, an ointment, implants (ear, eye, skin, nose, rectal and vaginal), intramammary preparations, vagitoria, suppositories, or intrauterine devices

It has been speculated that it is preferable to carry out a mucosal administration of a vaccine through the membrane mucosa, which undergoes natural exposure to the antigenic agent. Accordingly, for allergies for antigenic agents airborne mucosals, it is preferred to use administration by respiratory system medium, preferably an administration Oromucosal.

Correspondingly, for agent allergies mucosal antigens, which come into contact with the membrane digestive tract mucosa, it is preferred to use administration oral.

In an embodiment of the invention, the subject is submits to a vaccination protocol comprising administration Vaccine daily. In another embodiment of the invention the vaccination protocol comprises administration of the vaccine every Second day, every third day or every fourth day. For example, him vaccination protocol comprises administration of the vaccine over a period of more than 4 weeks, preferably more than 8 weeks, more preferably more than 12 weeks, more preferably more than 16 weeks, more preferably more than 20 weeks, more preferably more than 24 weeks, more preferably more than 30 and most preferably more than 36 weeks.

The administration period can be a continuous period Alternatively, the administration period is a discontinuous period interrupted by one or more periods of no administration. Preferably, the (total) period of no administration is shorter than the (total) period of administration.

In a further embodiment of the invention, the Vaccine is given to the test individual once a day. Alternatively, the vaccine is administered to the test individual. twice daily. The vaccine can be a single vaccine dose.

Oromucosal Preparation

Oromucosal administration can lead to out using any formulation of oromucosal administration available, including a solution, a suspension, forms fast dispersion pharmaceuticals, drops and pills for suck.

In a preferred embodiment of the invention, use sublingual immunotherapy (SLIT), in which case they are Preferred formulations fast dispersion pharmaceutical forms, drops and lozenges.

Examples of fast dispersion pharmaceutical forms are those described in US-A-
5,648,093, WO 00/51568, WO 02/13858, WO99 / 21579, WO 00/44351, US-A-4,371,516 and EP-278 877, as well as in the documents in process together with this document DK PA 203 00279 and DK PA 2003 00318 filed on behalf of the ALK-Abelló A / S assignee. Preferred rapid dispersion pharmaceutical forms are those produced by freeze drying. Preferred matrix forming agents are fish gelatin and modified starch.

The present invention can be used in desensitization of classical incremental dosing, in which the dose of allergen in the form of a solid pharmaceutical form of Fast dispersion is increased to a certain maximum. The power preferred of a unit dose of the pharmaceutical form is from 150-1000000 SQ-u / form pharmaceutical, more preferred the potency is from 500-500000 SQ-u / pharmaceutical form and more preferably the power is from 1000-2500000 SQ-u / form pharmaceutical, even more preferred 1500-125000 SQ-u / pharmaceutical form most preferable 1500-75000 SQ-u / form Pharmaceutical

In another embodiment of the invention the form pharmaceutical is a repeated single dose, preferably within the range of 1500-75000 SQ-u / form Pharmaceutical

Sensitization

The subject to be treated is sensitized as to present a specific IgE response to the administered allergen. In connection with the present invention the expression "present a allergen-specific IgE response "means a level of allergen specific IgE antibody detectable in at least one immunoassay The detection of the specific IgE antibody of allergen can be carried out using any immunoassay conventional, for example those described in WO documents 94/11734 and WO 99/67642.

In a particular embodiment of the invention the subject is further sensitized to present an answer of allergen-specific Th2 lymphocytes.

In a particular embodiment of the invention the subject is further sensitized to present an answer specific allergen test on a Simple Puncture Test Cutaneous (SPT).

In a further particular embodiment of the invention the subject is less than 40 years old, preferably of less than 30 years, more preferably less than 20 years and most preferably between 2 and 10 years old.

Clinical symptoms

The subject to be treated is symptom free allergy clinics associated with the allergen.

The associated allergy clinical symptoms with the allergen they can be any conventional symptom, including rhinitis, conjunctivitis, rhinorrhea, nasal obstruction, sinusitis, sneezing, atopic dermatitis, itching, watery eyes, runny nose, wheezing and skin irritation.

A number of factors are indicative for the development of allergy with clinical symptoms manifested later in the life. Hereinafter the subjects presenting one or more of such Indicator factors are referred to as high-risk subjects. The Indicators of high-risk subjects are clinical symptoms of allergies associated with one or more allergens other than vaccine allergen Additional indicator factors of subjects High risk are the presence of one or more allergies in one or both parents or grandparents or in one or more brothers or sisters. The treatment preventive according to the invention is particularly suitable for high risk subjects. However, the subject to be treated may also be a subject that has no indicator factor of high-risk subjects, for example free of clinical symptoms of Allergy to other allergens.

Formulation of the allergy vaccine

The allergy vaccine used in the invention may be in the form of any suitable formulation for administration to a mucosal surface, including a sprayer, a spray, a mixture, tablets (enterorrecovered and not enterorrecovered), capsule (hard and soft, enterorrecubierta and not enterorrecubierta), a suspension, a dispersion, granules, a powder, a solution, an emulsion, chewable tablets, drops, a gel, a paste, a syrup, a cream, a lozenge (powder, granules, tablets), a rapid dispersion tablet, an instillation fluid, a gas, a steam, an ointment, a bar, implants (ear, eye, cutaneous, nose, rectal and vaginal), preparations intramammary, vagitoria, suppositories or devices intrauterine

It is also understood that the vaccine of the invention may additionally comprise additional adjuvants and other excipients suitable for such type of formulation. Such Additional adjuvants and excipients are well known by the person skilled in the art and include among other solvents, emulsifiers, wetting agents, plasticizers, substances dyes, viscosity adjusting agents, buffering agents, mucoadhesive substances and the like. Examples of formulation strategies by the person skilled in the technique.

Adjuvant

The mucosal allergy vaccine may include a adjuvant, which can be any conventional adjuvant, including metal salts that contain oxygen, labile enterotoxin Heat (LT), cholera toxin (CT), toxin subunit B choleric (CTB), polymerized liposomes, mutant toxins, by example LTK63 and LTR72, microcapsules, interleukins (for example IL-1?, IL-2, IL-7, IL-12, IFNγ), GM-CSF, MDF derivatives, CpG oligonucleotides, LPS, MPL, phosphoacenes, Adju-Phos®, glucan, antigen formulation, liposomes, DDE, DHEA, DMPC, DMPG, DOC / Alum Complex, Freund's incomplete adjuvant, ISCOM®, Oral LT adjuvant, muramyl dipeptide, monophosphoryl lipid A, muramyl tripeptide and phosphatidylethanolamine.

The metal salt that contains oxygen can be any salt containing oxygen that provides the effect wanted. In a preferred embodiment, the metal salt cation which contains oxygen is selected from Al, K, Ca, Mg, Zn, Ba, Na, Li, B, Be, Fe, Si, Co, Cu, Ni, Ag, Au and Cr. preferred embodiment, the anion of the metal salt containing oxygen is selected from sulfates, hydroxides, phosphates, nitrates, iodates, bromates, carbonates, hydrates, acetates, citrates, oxalates and tartrates and mixed forms thereof.

Examples are aluminum hydroxide, phosphate aluminum, aluminum sulfate, potassium aluminum sulfate, phosphate of calcium, Maalox (mixture of aluminum hydroxide and hydroxide of magnesium), beryllium hydroxide, zinc hydroxide, carbonate zinc, zinc chloride and beryllium sulfate.

Allergy shots in the form of a solution aqueous or fast dispersion tablet, see document WO 04/047794, are particularly suitable for administration Oral and sublingual.

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Definitions

In connection with the present invention, they are used The following definitions:

The term "oromucosal administration" makes reference to a route of administration in which the form pharmaceutical is placed under the tongue or anywhere else apart from there in the oral cavity (oral administration) to allow the active ingredient to come into contact with the membrane mucosa of the oral cavity or pharynx of the patient in order to obtain a local or systemic effect of the active ingredient. A example of an oromucosal route of administration is the sublingual administration

The term "sublingual administration" makes reference to a route of administration, in which a pharmaceutical form under the tongue in order to obtain a local or systemic effect of the active ingredient.

The term "SQ-u" means SQ-Unit: the SQ-Unit is determined according to the standardization procedure of "biopower of SQ" by ALK-Abelló A / S, where 100,000 units of SQ equal the maintenance dose standard subcutaneous. Normally 1 mg of extract contains between 100,000 and 1,000,000 SQ-Units, depending on the source of allergen from which it originates and the procedure of elaboration used. The precise amount of allergen can be determine by means of immunoassay that is allergen content Total principal and total allergenic activity.

The expression "immunomodulatory treatment" it means that the treatment modulates the immune response of the treated subject.

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Examples Example 1 The effect of SLIT in a mouse model of Rhinitis Basis

The rhinitis model was projected in order to test the effect of sublingual immunotherapy (SLIT) in a model of mouse with clinical manifestations.

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Procedures Animals

Female BALB / c mice 6-10 weeks old were raised in the laboratory of the authors of the present invention and maintained on a defined diet that did not contain cross-reaction components with antisera against Phleum pratense (Phl p). Each experimental group consisted of 8-10 animals.

Animal experiments

The mice were sensitized with three Intraperitoneal (ip) injections of Phl p extract adsorbed to alum, followed by sublingual treatment with Phl p o extract buffer for 6-9 weeks. Mice are underwent provocation test intranasally for two weeks with Phl p extract and analyzed for symptoms clinical as described above. After the sacrifice collected for analysis blood, bronchoalveolar fluid (BAL), nasopharyngeal fluid (NAL), spleen and cervical lymph nodes.

Clinical Data

Sneezing and nasal rubbing: mice were observed within 10 minutes after administration intranasal of Phl and the number of sneezes and of nose rubs.

Hyperreactivity of the respiratory tract: using a full-length plethysmograph induced obstruction of the airways by increasing concentrations of aerosolized methacholine. Respiratory flow obstruction was measured Potentiated pulmonary pause (Penh) over a period of 6 minutes after administration of methacholine.

IgA Assay

Estapore magnetic beads were incubated (Stamp IB-MR / 0.86) coupled to anti-mouse IgA goat with BAL or NAL. Then washing and incubation were carried out. with biotinylated allergen. After washing was carried out incubation with streptavidin labeled LITE reagent, and after washing, light luminescence was measured on a luminometer (Magic Lite EQ Analyzer).

IgE Assay

Estapore magnetic beads were incubated (Stamp IB-MR / 0.86; A0201) coupled to IgE mouse mouse anti-mouse After washing was carried out incubation with biotinylated allergen. Then they were carried out washing and incubation with streptavidin labeled LITE reagent, and after washing, luminescence of light was measured in a luminometer (Magic Lite EQ Analyzer).

IgG, IgG1 and IgG2 assay

Phl p extract (10 µg / ml) was added to the wells of an ELISA plate and the plates were allowed to remain until the next day at 4-8 ° C. The coated plates were then washed with a buffer and blocked with a 2% casein buffer for one hour at room temperature on a stir table. After removing the casein buffer, the diluted serum sample was added to the plate and incubated at room temperature for two hours on a stir table. The plates were washed and biotinylated rabbit anti-mouse IgG / IgG1 / IgG2 diluted in 0.5% BSA buffer was added to each well and allowed to remain at room temperature for one hour on a stir table . After washing, streptavidin-HRP diluted in 0.5% BSA buffer was added to each well and allowed to stand at room temperature for one hour on a stir table. The plates were developed with TMP substrate (3,3 ', 5,5'-tetramethylbenzidine) for 20 minutes and stopped with 0.5 M H2SO4. The resulting reaction mixtures were subjected to a endpoint spectrophotometric measurement of
450 nm

T lymphocyte proliferation assay

The spleens were spun until suspension was removed Single cell and washed three times in the middle. Lymphocytes they were counted and adjusted to 1.67 x 10 6 cells / ml. Be 3 x 10 5 lymphocytes from a plate were added to each well 96-well flat bottom culture and lymphocytes are stimulated by 0, 10 and 40 µg / ml of Phl p extract. The Lymphocytes were cultured for 6 days at 37 ° C and at 5% CO2. Proliferation was measured by adding 0.5 µCi of 3 H-thymidine to each well during the last 18 hours of the cultivation period, followed by the collection of lymphocytes and the incorporated radiolabel count.

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Results Rhinitis induction

As will be apparent from Fig. 1A-E, the mice that were tested provocation intranasally (IN) for two weeks with extract Phl p (5000 SQ / mouse per day) after sensitization Intraperitoneal showed clear signs of rhinitis. First, they sneezed significantly more than mice sensitized undergoing provocation test with buffer (Fig. 1A). Second, they had elevated levels of IgE both in serum, as in BAL as in NAL (Fig. 1B-D). In third, these mice had an inflow of Eosinophil granulocytes increased in NAL (Fig. 1E).

The effect of SLIT in a mouse model of rhinitis

In order to assess the effect of SLIT on the previously described murine model of rhinitis, mice sensitized were treated sublingually for 9 weeks well with Phl p extract of 125,000 SQ or with pre-tested buffer of intranasal provocation with Phl p. In addition a group of mice sensitized did not undergo sublingual treatment, and were underwent provocation test intranasally with buffer serving as well as a negative control.

As shown in Figure 2, the treatment of SLIT was able to reduce clinical symptoms, since the number of Sneezing decreased significantly compared to mice treated with buffer (Figure 2A). Similarly, the hypersensitivity of the airways in response to proof of provocation with methacholine (as measured by Penh values) especially when mice were tested with doses between 5 and 15 mg / ml (Fig. 2B).

In addition, serum antibody levels are were affected by the treatment of SLIT. As seen in the Figure 3, serum levels of both Phl p specific IgE (Fig. 3A) and IgG1 (Figure 3B) were significantly reduced in mice treated with SLIT compared to mice treated with tampon.

Specific IgE levels decreased also in both BAL fluids and NAL fluids of mice treated with SLIT, while specific IgA levels are reduced in BAL fluids but not significantly in fluids NAL (see Fig. 4A-C).

In addition, the results indicate that the levels of eosinophil granulocytes in both BAL and NAL were reduced significantly in mice treated with SLIT compared to mice treated with buffer (see Fig. 5A-B).

Finally, the ex vivo reactivity of spleen lymphocytes and lymph node lymphocytes cervical (LN) (draining the tongue) after restimulation with Phl extract p. Although there was a tendency to regulate the low Phl p specific T lymphocyte response in the spleen, This was not statistically significant (Figure 6A). In In contrast to this, the T lymphocyte response to Phl p in LN of cervical drainage (Fig. 6B) was significantly decreased in mice treated with SLIT compared to mice treated with tampon.

conclusion

The results show that treatment with SLIT is able to reduce both clinical and clinical characteristics. the serological and cellular disease in a murine model of rhinitis.

Claims (18)

1. An allergen to use in treatment allergy prevention by administering an allergy vaccine that comprises the allergen to a mucosal surface of a subject,

to)

at that the subject to be treated is sensitized as to present an allergen-specific IgE response,

b)

at that the subject to be treated is free of allergy clinical symptoms associated with the allergen, and

C)

at that preventive treatment is aimed at avoiding or reducing symptoms subsequent allergy clinics associated with the allergen

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2. The allergen according to claim 1, in the that the subject is sensitized to present a response from Allergen-specific Th2 lymphocytes. 3. The allergen according to claim 1 or 2, in which the subject is free of clinical symptoms of rhinitis, conjunctivitis, rhinorrhea, nasal obstruction, sinusitis, sneezing, atopic dermatitis, itching, watery eyes, runny nose, wheezing and skin irritation. 4. The allergen according to any of the claims 1-3, wherein the subject is of less than 40 years, preferably less than 30 years, more preferably less than 20 years and most preferably between 2 and 10 years old. 5. The allergen according to any of the claims 1-4, which is selected from group consisting of an inhalation allergen and an allergen of poison. 6. The allergen according to claim 5, which is selected from the group consisting of a pollen allergen of trees, a pollenic allergen of grasses, a pollen allergen of dust mites, an allergen from herbaceous plants and an allergen animal. 7. The allergen according to any of the claims 1-6, wherein the administration it is oral administration by the mucous membrane of the system digestive, nasal, vaginal, sublingual, ocular, administration rectal, urinary, intramammary, pulmonary, otic or buccal. 8. The allergen according to claim 7, in the that the administration is oral or sublingual. 9. The allergen according to any of the preceding claims, in which the subject to be treated has clinical symptoms of allergies associated with one or more allergens other than the vaccine allergen. 10. Use of an allergen in the preparation of a allergy vaccine that comprises the allergen as a substance active for the allergic preventive treatment administering the vaccine against allergy to a mucosal surface of a subject,

to)

at that the subject to be treated is sensitized as to present an allergen-specific IgE response,

b)

at that the subject to be treated is free of allergy clinical symptoms associated with the allergen, and

C)

at that preventive treatment is aimed at avoiding or reducing symptoms subsequent allergy clinics associated with the allergen

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11. The use according to claim 10, wherein the subject is sensitized to present a response from Allergen-specific Th2 lymphocytes. 12. The use according to claim 10 or 11, in that the subject is free of clinical symptoms of rhinitis, conjunctivitis, rhinorrhea, nasal obstruction, sinusitis, sneezing, atopic dermatitis, itching, watery eyes, runny nose, wheezing and skin irritation. 13. Use according to any of the claims 10-12, wherein the subject is of less than 40 years, preferably less than 30 years, more preferably less than 20 years and most preferably between 2 and 10 years old. 14. Use according to any of the claims 10-13, wherein the allergen is selected from the group consisting of an inhalation allergen and a poison allergen 15. The use according to claim 14, wherein the allergen is selected from the group consisting of an allergen tree pollen, a grass pollen allergen, an allergen Pollinic of dust mites, an allergen from herbaceous plants and a animal allergen 16. Use according to any of the claims 10-15, wherein the administration it is oral administration through the mucous membrane of the system digestive, nasal, vaginal, sublingual, ocular, administration rectal, urinary, intramammary, pulmonary, otic or buccal. 17. The use according to claim 16, wherein administration is oral administration or administration sublingual 18. Use according to any of the claims 10-17, wherein the subject to be treated have clinical symptoms of allergies associated with one or more allergens other than the vaccine allergen.