EP1414489A2

EP1414489A2 - Antiallergic pharmaceutical composition

Info

Publication number: EP1414489A2
Application number: EP02757745A
Authority: EP
Inventors: Emile Loria; Ga[Tan Terrasse; Yves Trehin
Original assignee: Antialis
Current assignee: Antialis
Priority date: 2001-03-30
Filing date: 2002-03-28
Publication date: 2004-05-06
Also published as: AU2002308044A1; US20030104013A1; FR2822709B1; EP1473041A3; WO2002078736A2; ATE342732T1; BR0208710A; DE60215548D1; US7048928B2; EP1473041B1; FR2822709A1; WO2002078736A3; EP1473041A2

Abstract

Antiallergic compositions containing (i) an anti-histamine, (ii) a histamine synthesis inhibitor, and optionally (iii) an allergen or isolated nucleic acid molecule that has at least one polynucleotide sequence coding for the allergen, together with a pharmaceutical carrier. ACTIVITY : Antiallergic; Antiinflammatory; Antiasthmatic; Dermatological. MECHANISM OF ACTION : Prevention of the synthesis of Th2 cells from Th0 cells, thus blocking the synthesis of Interleukins 4 and 13.

Description

ANTI-ALLERGIC PHARMACEUTICAL COMPOSITION

The present invention relates to new pharmaceutical compositions for the prevention and treatment of allergy. Allergies constitute a plague from which 25% of the world population suffers. This number is increasing in connection with a change in the toxicity of the environment (dust, food, motor vehicle). In addition, the risk for an individual to suffer from an allergy increases with an allergic history in the parents.

The biological mechanism of the allergy can be described as an abnormally amplified reaction following the entry of the allergen into the body. The following events are responsible for the reaction: identification of the allergen by the body, secretion of cytokines in response to the penetration of allergens, stimulation of ThO cells favoring an accelerated differentiation of ThO cells into Th2 instead of their differentiation normal balanced in Thl and Th2 Th2 cells synthesize Interleukins 4 and 13, responsible for the worsening of allergic symptoms by the resurgence of IgE synthesis, the final phase of the reaction is the release of histamine and serotonin with a recruiting effect on the Th2 clones, a toxic and inflammatory self-maintenance reaction, even without antigen stimulation.

Antigen presenting cells (APCs: macrophages, dendritic cells, B lymphocytes) participate in the hypersensitivity reaction by fundamental cellular cooperation to cause the immune reaction. Allergy belongs to the class of defense against self. The main allergens are dust mites (80%) and pollens (20%).

The self-stimulation reactions of CPA clones have an effect on the general level of release of histamine and serotonin with the consequence of worsening. general clinical symptology.

The level of recruitment of new IgE secreting cells is thus increased, thereby facilitating the explosion of clinical signs when a new allergen enters the body. This can be observed in atopic patients where allergic reactions are violent due to a high level of Th 2 clones promoting the synthesis of IgE.

The observed overall reaction due _to the penetration of new allergen is not due to the toxicity of it, but just the fact that there is a trigger level phenomena very low allergic favored by other sensitizations.

For allergists, the allergy is commonly perceived as a reaction due to hypersynthesis of the immunoglobulin IgE. The inflammatory reaction mainly affects the respiratory and ENT spheres, with a pathological focus on the nose, lungs and skin. The pathologies associated with allergy are disabling and suffer from the lack of effectiveness of conventional treatment. There is no preventive strategy and the curative means are insufficient or badly used.

The usual treatment of allergic disease consists, firstly, in the recognition of the responsible allergen: mites, pollen, molds, food; secondly, eviction measures are taken, so as to avoid exposure of the subject to this (these) allergen (s); thirdly, a treatment is proposed whose focus is on the target organ which appears symptomatic: ENT treatment on rhinitis, anti-asthma treatment if the affected area is the respiratory area, dermatological treatment if the attack is cutaneous.

If the above measures fail, isolated or complementary therapeutic measures can be proposed. Generally, antihistasics are prescribed, but this treatment is not enough to make all the symptoms of the allergy go away. However, it is useful for reducing its intensity and making it more bearable for the patient. Also, immunotherapy is generally offered. This immunotherapy is based on the identification of the allergen, and its aim is to subject the subject gradually to higher and higher rates of the specific allergen (specific pollen, specific mite, specific molds) considered, so as to "Accustom" the body to the presence of this allergen. The complexity of the processing started prevents adhesion to the latter. The succession of therapies is a factor of obvious failure of the treatments. The pathologies associated with allergy are disabling and suffer from the lack of effectiveness of conventional treatments.

The inventors refined the knowledge of the biological mechanism of allergy by noting that the synthesis of excess IgE immunoglobulins was, upstream, caused by the increase in the level of Th2 cells and the disruption of ThO differentiation in favor of Th2. . Contrary to the usual approach of those skilled in the art, the inventors considered the allergy not as a specific reaction to a specific antigen, but as a global immune disorder due to the increase in Th2.

The technical problem that the inventors then identified consisted in knowing how to avoid this imbalance in favor of the synthesis of Th2 cells; how to block the imbalance in the preferred differentiation of ThO cells into Th2 cells. The solution proposed by the present invention aims to offer new means of treatment of allergies both preventive and curative.

This goal is achieved by treating the two main aspects of the immune reaction: on the one hand, the upstream of the immune reaction, which after presentation of the antigen to the CPA leads to an increased synthesis of IgE responsible for the self-recruitment of the cells of immunity and - on the other hand, the downstream of the immune reaction, which leads to the release of the preformed mediators, essentially histamine, responsible on the one hand for the final clinical symptomatology and acting on the other hand in stimulating the transformation of ThO into Th2.

The treatment according to the invention has the advantage of being non-specific of the allergen which is at the origin of the allergy and makes it possible to approach in a global manner the allergic disease without being concerned with the specificity of the allergen. In fact, the compositions according to the invention make it possible to treat a level of immune reactivity and not to offer specific immunotherapy. These compositions are effective for treating subjects for whom the allergy is clearly of a hereditary nature. The goal is to restore the body to a silent defense homeostasis in relation to its environment.

The invention relates to an anti-allergic pharmaceutical composition comprising (i) an antihistamine compound, (ii) an inhibitor of histamine synthesis, and optionally (iii) an allergen or an isolated nucleic acid molecule comprising at least one sequence polynucleotide coding for said allergen, these being combined in said composition with a pharmaceutically acceptable vehicle.

According to a particular embodiment, the invention relates to an anti-pharmaceutical composition. allergic comprising at least one antihistamine compound and at least one inhibitor of histamine synthesis, said compounds being combined in said composition with any pharmaceutically acceptable vehicle.

Advantageously, the antihistamine agent is chosen from the group comprising: brompheniramine, cetirizine, f exofenadine, cyproheptadine, dexchlorpheniramine, hydroxizine, ketotifene, loratadine, mequitazine, oxoto ide, izolastine , ebastine, 1 ast émi z oie, carbinoxamide,

1 alimemazine, buclizine, cyclizine hydrochloride, doxylamine.

Advantageously, the inhibitor of histamine synthesis is an inhibitor of histidine decarboxylase, preferably tritoqualine.

The preferred amounts of antihistamine compound in the composition according to the invention are between 1 and 2000 mg and preferably between 5 and 200 mg, and the preferred amounts of inhibitor of histamine synthesis are between 1 and 2000 mg.

According to a particularly advantageous embodiment, the composition according to the invention comprises 5 to 200 mg of antihistamine, and from 10 to 300 mg of histidine decarboxylase inhibitor.

According to a variant of the invention, the composition also comprises an allergen, chosen from the allergens or mixture of major allergens of mites capable of inducing an immune reaction, in particular of mites D. Pteronyssinus and / or D. Farinae. Preferably, this allergen is a cystine protease, or at least a peptide epitope of a cystine protease. Of preferably, it is a peptide epitope of a cystine protease whose amino acid sequence comprises the sequence SEQ ID NO: 2 in the sequence list in the appendix. According to another embodiment of the invention, the allergen is a peptide or a mixture of peptides chosen from the group comprising the peptides of sequences SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 in the attached sequence list.

The allergen used in the combination of the invention can be natural and obtained by extraction of D mites. Pt eronyssinus and / or D. Farina e. Alternatively, it can be synthetic and obtained by peptide synthesis according to conventionally known techniques. Preferably, the amounts of allergen present in the composition of the invention are of the order of 1 to 1500 μg and preferably from 10 to 150 μg.

Advantageously, the isolated nucleic acid molecule comprising at least one polynucleotide sequence coding for said allergen, is a vector, in particular an adenoviral vector. Advantageously, the polynucleotide sequence coding for the allergen contained in the composition of the invention, codes at least for a cystine protease of sequence SEQ ID NO: 2 in the list of sequences in the appendix or, according to a variant of the invention, at least for the three peptide epitopes SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 in the sequence list in the appendix. Advantageously, this nucleotide sequence comprises a sequence corresponding to the sequence SEQ ID NO: 1 in the list of sequences in the appendix, or, according to a variant of the invention, at least one of the sequences SEQ ID No.: 6 or SEQ ID No.: 7 in the attached sequence list.

According to a preferred embodiment, the composition according to the invention is released in the form of a transcutaneous patch to allow better access of the allergens used and / or of the polynucleotide sequences coding for said allergens, to the antigen presenting cells.

According to an alternative embodiment, the composition according to the invention is released in the mucosal form, eye drops, nasal or bronchial spray.

According to another variant, the composition according to the invention is released in galenical form with disintegration programmed in mucosal or sub-lingual and secondarily per os.

The pharmaceutical composition of the invention is useful for the preparation of a medicament for the treatment or prevention of the allergic hypersensitivity reaction. The composition according to the invention is also useful for the preparation of a medicament intended for the treatment or prevention of allergic asthma, allergic rhinitis, atopic and allergic eczema The composition according to the invention is still useful for the preparation of a medicinal product intended for the treatment or prevention of allergic manifestations in children, infants and adults.

The invention also relates to the use of a first allergen or of an isolated nucleic acid molecule comprising at least one polynucleotide sequence coding for said first allergen, for the preparation of a pharmaceutical composition. useful for treating or preventing an allergy caused by a second allergen different from the first. A first preferred allergen is a mite cystine protease. According to a preferred embodiment of the invention, the first allergen is a cystine protease from a mite and the other allergen is not a cystine protease from a mite.

The invention will be better understood on reading the detailed description below, which illustrates, without limitation, the invention.

The inventors based their first experiments on the toxic and inflammatory terminal reaction of the allergy, which self-maintains in the absence of antigenic stimulation. This reaction is usually treated by blocking the terminal phase of the allergy, by administration of an antihistamine, which acts competitively with histamine for binding to histamine receptors. Blocking histamine receptors is the main effector pathway. This blocking must be carried out over a sufficiently long time so that a negative feedback on the synthesis of these receptors is also carried out. Antihistamines are the anti-receptor molecules of choice to block this terminal reaction.

As mentioned above, the allergy is also accompanied by an increased synthesis of histamine which is also a cause of self-maintenance of the terminal inflammatory reaction. This synthesis of histamine can optionally be controlled to improve the effectiveness of the pharmaceutical composition proposed previously. This control involves inhibition histamine synthesis, in particular by means of a histidine decarboxylase inhibitor. By preventing the synthesis of histamine, the histidine decarboxylase inhibitor enhances the effectiveness of the composition in its action on the downstream phase of the biological mechanism of the allergy by complementing the antihistamine part. This is why the preferred compositions of the invention comprise an effective amount of an antihistamine compound associated with an inhibitor of the synthesis of histamine. Thus the blocked effector histamine terminal pathways will allow effective control of the cascade of the allergic reaction. The terminal pathway for synthesis and stimulation of histamine receptors is thus blocked overall.

The inventors then found that the administration of an allergen, in combination with an antihistamine and an inhibitor of the synthesis of histamine, gave surprisingly effective results and they developed the compositions according to the invention which provide a new allergenic approach constituting a preventive vaccination and not specific to the development of allergic diseases. Advantageously, this allergen is chosen from the antigens or mixture of major mite antigens capable of inducing an immune reaction. In fact, the work carried out within the framework of the invention consisted in identifying and then using ubiquitous antigens of mites.

These antigens are present in large quantities in the environment and are at the origin of the development of allergic reactions in the world. Two mites, D. Pteronyssinus (DP) and D. Farinae (DF) are the most represented in the global environment.

The invention envisages very particularly as an allergen, a cystine protease carrying the antigenicity which is identical to 90% for these two mites. The amino acid sequence of the cystine protease of D. Pteronyssinus (DP) is represented in the sequence list in the appendix respectively under the number SEQ ID NO: 2. The allergens used in the compositions of the invention can be, either extracts obtained from raw biological material, or totally or partially purified proteins, possibly produced by genetic engineering, or by peptide synthesis.

According to a particular embodiment of the invention, the composition comprises, as an allergen, peptide epitopes of cystine protease. It has indeed been highlighted three epitopic parts which are identified as being triggers of the immune reaction. These are the three peptides with the following sequences:

RMQGGCGSCN (SEQ ID NO: 3)

QPNYHAVNIV (SEQ ID NO: 4) TVRNSWDT (SEQ ID NO: 5) and their possible analogs.

The protein epitope sequences mentioned above may contain additional amino acid sequences or substitutions facilitating their attachment to the Major Histocompatibility Complex (MHC). The invention therefore especially envisages pharmaceutical compositions comprising as allergen at least one of these three peptides.

These peptide epitopes are strictly identical in DF, DP, as in other mites, because they carry the enzymatic function of cystine protease. Their lipophilicity, as well as the fact that they support the enzymatic function, explain why these epitopic parts are constant from one species of mites to another and the inventors consider that they are at the origin of a general immune reaction. .

The use of these peptides, either in cyclized form, or in epigenic form, even in the form of a nucleic acid (DNA or RNA) comprising a nucleotide sequence coding for at least one of these peptides, must induce tolerance to the natural antigen and decrease the general level of the upstream immune reaction.

The cyclization of the peptides and / or their inclusion in a larger sequence allows an improvement in the presentation of the antigens to the T lymphocytes. This improvement in the presentation will allow a presentation of the antigens and the epitopes at the MHC and by this will trigger the immune reaction. of tolerance. Indeed, the antigens must first be reworked by the CPA. The simple epitopic form does not allow reworking by the CPA because, in general, only a protein longer than 10 amino acids can be cut and presented by the CPA to the T lymphocytes. These peptides can be associated with any pharmaceutically acceptable vector, for example of a phospholipid nature.

In the case of nucleic acids, the sequences coding for said peptides can be amplified by the following nucleotide sequences:

5'GCGGCGGCG 3 '(SEQ ID NO: 6) or 5' TGAGCGGCGGCG 3 '(SEQ

ID NO: 7).

It is thus possible to integrate the epigens corresponding to the DP / DF epitopes with a sequence of nucleotide primers of sequence (SEQ ID NO: 7) by alternating said sequence (SEQ ID NO: 7) and an epitope so as to integrate the three major epitopes of DP / DF taken together or separately. The integration of the epitopes taken together leads to having a set consisting of a nucleotide primer sequence (SEQ ID NO: 7) a first major epitope, a nucleotide primer sequence (SEQ ID NO: 7), a second major epitope, a nucleotide primer sequence (SEQ ID NO: 7), a third major epitope.

The integration of the epitopes taken separately leads to mixing three sets each consisting of a sequence of nucleotide primers (SEQ ID NO: 7) and a major epitope. This integration of the epitopes with a nucleotide primer sequence according to the following sequence (SEQ ID NO: 7) should improve the efficiency of presentation of the DP / DF epigens to T lymphocytes. By this improved presentation, the epigens of, DP / DF will stimulate the TH1 switch and therefore lower the level of allergic reaction.

The use of these epitopes, or a solution allowing the TH1 / TH2 switch such as primers nucleotides according to the sequence (SEQ ID NO: 7) on the one hand, and their association with an antihistamine compound and possibly an inhibitor of the synthesis of histamine, on the other hand, constitute an effective and innovative solution for prevention and treatment allergy.

Consequently, the compositions of the invention comprise an effective amount of at least one allergen as defined above without foreshadowing the role of this allergen in the symptomatology of the patient.

This approach allows a comprehensive approach to allergic disease without worrying about the specificity of the allergen. In fact, the composition according to the invention makes it possible to treat a level of immune reactivity and not to offer specific immunotherapy.

The use of the allergen, in the various forms described above, in the compositions according to the invention, makes it possible to induce tolerance to the natural antigen and reduces the general level of the upstream immune reaction.

The compositions according to the invention comprise an amount of allergens of the order of 1 to 1500 μg and advantageously from 10 to 150 μg. In the case of peptides, each of these is advantageously present in proportions of the order of 1 to 1500 μg so as to slow down the immunological reaction leading to increased synthesis of IgE.

The ... compositions according to the invention may be in a form for transdermal application, for example an ointment for children, for oral administration, for example a slow-release lyoc, or alternatively gastro-tablets. resistant or erasers. It can also be a spray or eye drops, or galenical forms with disintegration programmed in mucosal and secondarily per os.

Thus, the compositions of the invention lend themselves to different modes of administration chosen in adaptation to the pathological profile of the patient and his age. For children, the patch form or the syrup or sucking tablet form. The other eye drops or injection forms can also be used. In adults, all dosage forms are possible.

The advantage of a coupled form also allows therapeutic simplicity, adherence to simplified treatment and therefore ultimately better treatment success.

This solution also makes it possible to prevent allergic disease and not just overt medical conditions. Children of allergic parents could be the major target of these preventive treatments. The result is less hospital stay, less antibiotic treatment and an improved quality of life.

Indeed, the rebalancing of the differentiation of ThO cells into Thl and Th2 cells must take place as early as possible to be effective, because in infants it is the differentiation in favor of TH2 cells which is predominant, responsible for hyper-reactivity at l 'environment. The rebalancing TH2 / TH1 must be early to be as durable as possible because antigenic stimulation by environmental antigens (mites, and bacteria) are stimulators of the TH2 pathway. Thus, the pharmaceutical composition according to the invention is particularly useful for the preparation of a medicament intended for the treatment of the allergic hypersensitivity reaction. Advantageously, the pharmaceutical composition according to the invention is in a galenical form with disintegration programmed in mucosal or sub lingual and secondarily per os.

The pharmaceutical composition according to the invention is also useful for the preparation of a medicament intended for the treatment or prevention of the allergic hypersensitivity reaction, for the treatment or prevention of allergic asthma of allergic rhinitis of the atopic and allergic eczema. Finally, the pharmaceutical composition according to the invention is particularly useful for the preparation of a medicament intended for the treatment or prevention of allergic manifestations in children, infants and adults.

Other advantages and characteristics of the invention will appear on reading the clinical observations concerning the treatment of allergic patients according to the table cited below. These observations were made on a hundred patients, who were administered a composition according to the invention combining at least one antihistamine compound, a histamine synthesis inhibitor and an allergen. The patients are between 7 and 60 years old. They present at least one positive mite or pollen test, tested by prick test, as well as a symptomatology of rhinitis or asthma for at least one year.

The pathological profile of the patients is classified according to the following typology comprising three descriptive categories: inflammation, secretion and the figurative element.

- Only the clinical examination can classify the inflammation. It is considered that there is inflammation when the examination of the mucous membranes or of the target organs shows a red coloration thus signifying an inflammatory phenomenon.

- Secretion concerns the observation of a purulent or non-purulent exudate which reaches a target organ (mucous membranes, skin, etc.). - The figured element is a modification of the structure of the organ considered which can appear in several pathological forms. We only take into account the presence of this modification without going into the details of this modification. The classification of pathological severity is done on 4 levels distinguishing the intensity of the impairment according to a classification ranging from 1 to 4, in the form of a fraction 1/4, 1/2 or a whole number.

Thus, according to this classification, a rating of 1/4 means an attack on the target organ between 0 and less than 1/4. For a rating of 1/2, this classification means that the damage is between 1/4 and half of the target organ; for 3/4, this notation means that. the impairment is greater than 1/2 and less than 3/4; for 1, it means that the damage is greater than 3/4. A first category of target organs is classified according to this typology. It includes the eyes, nose, pharynx, larynx and skin.

For the lungs, the classification is made according to the results of a respiratory functional exploration with figures expressed as a percentage compared to the normal value (according to an international classification taking account in particular of age and size).

Patients are followed up with visit at least 2 months, 8 months, 12 months, 24 months. The evolution of the therapies taken and the number of units taken are analyzed.

'Table I below gives a clear indication of the very positive results obtained after approximately 8 months of treatment. There is a clear improvement in the pathological condition of the patients with a total clinical severity score passing on average from an index 9.56 to an index 2.47 and a mean deviation evolving from 1.15 to 0.53 confirming the effectiveness of treatment on all age and sex groups of patients. The average number of target organs affected decreased from 3.69 to 1.73 while the average difference in the number of target organs affected was reduced from 0.49 to 0.41. Table I

Table II below shows the average of the clinical scores as well as the average difference of the scores obtained.

Table II

Table III below illustrates the average number of target organs affected as well as the average difference in the number of target organs reached. Table III

Claims

1) Anti-allergic pharmaceutical composition characterized in that it comprises: (i) an antihistamine compound, (ii) an inhibitor of histamine synthesis, and optionally (iii) an allergen or an isolated nucleic acid molecule comprising at at least one polynucleotide sequence coding for said allergen, these being associated in said composition with a pharmaceutically acceptable vehicle.

2) Anti-allergic pharmaceutical composition according to claim i, characterized in that it comprises at least one antihistamine compound and at least one inhibitor of histamine synthesis, said compounds being combined in said composition with any pharmaceutically acceptable vehicle .

3) Pharmaceutical composition according to any one of claims 1 or 2, characterized in that said antihistamine agent is chosen from the group comprising: brompheniramine, cetirizine, fexofenadine, cyproheptadine, dexchlorpheniramine, hydroxizine, ketotifen, loratadine, mequitazine, oxotomide, mizolastine, ebastine, astemizole, carbinoxamide, alimemazine, buclizine, cyclizine hydrochloride, doxylamine.

4) Pharmaceutical composition according to any one of claims 1 or 2, characterized in that said inhibitor of histamine synthesis is an inhibitor of histidine decarboxylase. 5) Pharmaceutical composition according to claim 4, characterized in that the inhibitory compound of hi st idine decarboxyla is tritoqualine.

6) Pharmaceutical composition according to any one of claims 1 to 5, characterized in that it comprises an amount of antihistamine compound between 1 and 2000 mg and preferably between 5 and 200 mg.

7) Pharmaceutical composition according to any one of claims 1 to 6, characterized in that it comprises an amount of inhibitor of the synthesis of histamine between 1 and 2000 mg.

8) Pharmaceutical composition according to any one of claims 1 to 7, characterized in that it comprises from 5 to 200 mg of antihistamine, and from 10 to 300 mg of a histidine decarboxylase inhibitor.

9) Pharmaceutical composition according to any one of claims 1 to 8, characterized in that said allergen is chosen from antigens or mixture of major antigens of mites capable of inducing an immune reaction.

10) Pharmaceutical composition according to any one of claims 1 to 9, characterized in that said allergen is a major antigen of D. Pteronyssinus and / or D. Farinae. 11) Pharmaceutical composition according to any one of claims 1 to 10, characterized in that the allergen is a cystine protease.

12) Pharmaceutical composition according to any one of claims 1 to 11, characterized in that the allergen is at least one peptide epitope of a cystine protease.

13) Pharmaceutical composition according to any one of claims 1 to 12, characterized in that the allergen is at least one peptide epitope of a cystine protease of sequence SEQ ID NO: 2 in the list of sequences in the appendix.

14) Pharmaceutical composition according to any one of claims 1 to 13, characterized in that the allergen is a peptide or a mixture of peptides chosen from the group comprising the peptides of sequences SEQ ID NO: 3, SEQ ID NO: 4 , SEQ ID NO: 5 in the attached sequence list.

15) Antiallergic pharmaceutical composition according to claim 1 to 14, characterized in that said allergen is natural and obtained by extraction of mites D. Pteronyssinus and / or D. Farinae.

16) Pharmaceutical composition according to any one of claims 1 to 15, characterized in that it comprises an amount of allergen of the order of 1 to 1500 μg and preferably from 10 to 150 μg. 17) Pharmaceutical composition according to any one of claims 1 to 9, characterized in that said isolated nucleic acid molecule comprising at least one polynucleotide sequence coding for said allergen, is an adenoviral vector.

18) Pharmaceutical composition according to any one of claims 1 to 9 and 17, characterized in that said polynucleotide sequence coding for said allergen, comprises a sequence SEQ ID NO: 1 in the list of sequences in the appendix.

19) Pharmaceutical composition according to any one of claims 1 to 9 and 17 to 18, characterized in that said polynucleotide sequence coding for said allergen comprises a nucleotide sequence corresponding to the sequence SEQ ID No.: 6 in the list of sequences in Annex.

20) Pharmaceutical composition according to any one of claims 1 to 9 and 17 to 19, characterized in that said polynucleotide sequence coding for said allergen and included in isolated nucleic acid molecule comprises a nucleotide sequence corresponding to the sequence SEQ ID No .: 7 in the attached sequence list.

21) Pharmaceutical composition according to any one of the preceding claims, characterized in that it is released in the form of a transcutaneous patch to allow better access to the allergens used and / or to the polynucleotide sequences coding for said allergens, to antigen presenting cells.

22) Pharmaceutical composition according to any one of the preceding claims, characterized in that it is released in the mucosal form, eye drops, nasal or bronchial spray.

23) Pharmaceutical composition according to any one of the preceding claims, characterized in that it is released in galenical form with programmed disintegration in mucosal or sub-lingual and secondarily per os.

24) Pharmaceutical composition according to any one of the preceding claims, for the preparation of a medicament intended for the treatment or prevention of the allergic hypersensitivity reaction.

25) Pharmaceutical composition according to any one of the preceding claims, for the preparation of a medicament intended for the treatment or prevention of allergic asthma, allergic rhinitis, atopic and allergic eczema

26) Pharmaceutical composition according to any one of the preceding claims, for the preparation of a medicament intended for the treatment or prevention of allergic manifestations in children, infants and adults.

27) Use of a first allergen or of an isolated nucleic acid molecule comprising at least at least one polynucleotide sequence coding for said first allergen, for the preparation of a pharmaceutical composition useful for treating or preventing an allergy caused by a second allergen different from the first.

28) Use according to claim 27 characterized in that said first allergen is a cystine protease of a mite.

29) Use according to claim 28 characterized in that said first allergen is a cystine protease of a mite, and in that said other allergen is not a cystine protease of said mite.