BRPI0618241A2 - use of an allergen to manufacture a liquid vaccine formulation for allergy prevention or treatment - Google Patents

Abstract

USE OF AN ALLERGEN TO MANUFACTURE A LIQUID VACCINE FORMULATION FOR THE PREVENTION OR TREATMENT OF ALLERGY The present invention relates to the use of an allergen to manufacture a liquid vaccine formulation for the prevention or treatment of allergy in an individual through oromucosal administration. in a dosage regimen not comprising progressive dosing.

Description

Report of the Invention Patent for "USE OF AN ALERGEN FOR THE MANUFACTURE OF A VACINALIGID FORMULATION FOR ALLERGY PREVENTION OR TREATMENT".

TECHNICAL FIELD

The present invention relates to an allergy vaccine formulation for oromucosal administration.

BACKGROUND OF THE INVENTION

Allergy is a major health problem in countries where the Western lifestyle is adapted. In addition, the prevalence of allergic disease is increasing in these countries. Although allergy in general cannot be considered a life-threatening disease, asthma causes a significant number of deaths annually. An exceptional prevalence of about 30% in adolescents leads to a substantial loss in quality of life, working days and money and ensures a ranking among the top health problems in the Western world.

Allergy is a complex disease. Many factors contribute to the awareness event. Among these is the susceptibility of the individual defined by a reciprocal action insufficiently understood between various genes. Another important factor is allergen exposure above certain thresholds. Several environmental factors may be important in the sensitization process including pollution, childhood infections, parasitic infections, intestinal microorganisms, etc. Once an individual is sensitized and the allergic immune response is established, the presence of only minute amounts of allergy is efficiently translated into symptoms.

The natural course of allergic disease is usually accompanied by worsening on two levels. First, a progression of symptoms and severity of the disease, as well as disease progression, for example from hay fever to asthma. Second, dissemination in offensive allergens occurs more frequently, resulting in allergic multi-reactivity. Chronic inflammation leads to a general weakening of mucosal defense mechanisms, resulting in non-specific irritation and eventually destruction of mucosal tissue. Babies may become primarily sensitized to food, ie milk, resulting in eczema or gastrointestinal disorders; more often, however, they develop these symptoms spontaneously. These babies are at risk of developing inhalation allergy later in their lives.

The most important sources of allergens are found in the most prevalent particles of a certain size in the air that they will breathe. These sources are markedly universal and include pollen pollen and household dust mite fecal particles, which together account for approximately 50% of all. the allergies. Of global importance are also animal dandruff, that is, cat and dog dandruff, other po-lens, such as sagebrush pollens, and microfungi, such as Alternaria. On a regional basis, still other pollens can dominate, such as northern and central European pollen, white yosna in the eastern and central United States, and Japanese cedar pollen in Japan. Insects, ie bee and wasp venom , and foods each account for approximately 2% of all allergies.

Allergy, ie, type I hypersensitivity, is caused by an appropriate immune response to foreign non-pathogenic substances. Important clinical manifestations of allergy include asthma, hay fever, eczema, and gastrointestinal disorders. The allergic reaction is prompt and peaks within 20 minutes of contact with the offending allergen. In addition, the allergic reaction is specific in that a particular individual is sensitized to a particular allergen (s), while the individual does not necessarily show an allergic reaction to other substances known to cause allergic disease. The allergic phenotype is characterized by pronounced inflammation of the target organ mucosa and the presence of the IgE class allergen-specific antibody in the circulation and on the surface of mast cells and basophils.

An allergic attack is initiated by the reaction of the foreign allergen with allergen-specific IgE antibodies when antibodies are bound to high affinity IgE-specific receptors on the mast cell and basophil surface. Mast cells and basophils contain preformed mediators, i.e. histamine, tryptase and other substances, which are released upon cross-linking of two or more re-receptor-linked IgE antibodies. IgE antibodies are cross-linked through the simultaneous ligation of an allergen molecule. Therefore, it follows that a foreign substance having only one antibody binding epitope does not initiate an allergic reaction. Cross-linking of receptor-bound IgE onto the mast cell surface also leads to the release of signaling molecules responsible for attracting eosinophils, allergen-specific T cells, and other types of cells to the site of the allergic response. These cells, in reciprocal effect with the allergen, IgE, and effector cells, lead to rapid symptom renewal, occurring 12-24 hours after encounter with a-allergen (late phase reaction).

Allergic disease management comprises diagnosis and treatment including prophylactic treatments. Allergy diagnosis refers to the demonstration of allergen-specific IgE and identification of the allergen source. In many cases a careful history may be sufficient for the diagnosis of allergy and for identification of the source material of the offending allergen. More often, however, the diagnosis is supported by objective measures, such as a skin prick test, blood test, or provocation test.

Therapeutic options fall into three main categories. The first opportunity is to avoid the allergen or reduce exposure. Although avoiding the allergen is obvious, for example, for food allergens, it can be difficult or expensive, as for house dust mite allergens, or it can be impossible, for pollen allergens. The second most widely used therapeutic option is the prescription of classic symptomatic drugs, such as antihistamines and steroids. Symptomatic drugs are safe and efficient; however, they do not alter the causal nature of the disease, nor do they control the spread of the disease. The third alternative therapy is allergy-specific vaccination which, in most cases, reduces or alleviates the allergic symptoms caused by the allergen in question.

Conventional allergy-specific vaccination is a ca-out treatment for allergic disease. It interferes with basic immunological mechanisms, resulting in persistent improvement of the patient's immune state. Thus, the protective effect of allergy-specific vaccination extends beyond the treatment period in contrast to symptomatic drug treatment. Some patients who receive treatment are cured, and in addition, most patients experience relief in the severity of the disease and symptoms or at least one interruption in the aggravation of the disease. Thus, allergy-specific vaccination has preventive effects, reducing the risk of development. hay fever in asthma and reducing the risk of developing new sensitivities.

The immune mechanism that underlies successful allergy-specific vaccination is not known in detail. A specific immunoresponse, such as antibody production against a particular pathogen, is known as an adaptive immune response. This response can be distinguished from the innate immune response, which is a nonspecific reaction to pathogens. An allergy vaccine is mentioned as referring to the adaptive immune response, which includes cells and molecules with antigen specificity, such as T cells and antibody producing B cells. B cells cannot mature in cells that "produce antibody without help from T cells of corresponding specificity. T cells that participate in the stimulation of allergic immune responses are primarily Th2 type. The establishment of a new balance between Th1 and Th2 cells has been proposed. as beneficial and central to the immune mechanism of allergy-specific vaccination.If this is accomplished through a reduction in Th2 cells, a Th2 cell shunt or Th1 overregulation is controversial. Recently, it has been proposed that T cells are According to this model, regulatory T cells, that is, Thh3 or Th1 cells, down-regulate Th1 and Th2 cells to antigen-specificity, despite these ambiguities, it is believed that generally, that an active vaccine should have the ability to stimulate allergen-specific T cells, preferably cell as TH1.

Allergy-specific vaccination, despite its virtues, is not widespread, primarily for two reasons. One reason is that the inconveniences associated with the traditional vaccination program include repeated vaccinations, ie injections over several months. The other reason is, more importantly, the risk of side allergic reactions. Common vaccinations against infectious agents are efficiently performed using one or a few high dose immunizations. This strategy, however, cannot be used for allergy vaccination as a pathological immune response is already underway.

Conventional allergy-specific vaccination, therefore, is performed using multiple subcutaneous immunizations applied over an extended period of time. The course is divided into two phases, the progressive dosing and the maintenance phase. In the progressive dosing phase, increasing doses are applied, typically over a period of 16 weeks, starting with small doses. When the recommended maintenance dose is reached, this dose is applied to the maintenance phase, typically with injections every six weeks. After each injection, the patient should remain under medical attention for 30 minutes because of the risk of anaphylactic side reactions which, although extremely rare, could in principle be life threatening. In addition, the clinician should be equipped to support emergency treatment. There is no doubt that a vaccine based on a different route of administration would eliminate or reduce the risk of inherent allergic side reactions in current subcutaneous-based vaccines, as well as facilitate widespread use, possibly even allowing self-vaccination at home.

Attempts to improve allergy-specific vaccines have been undertaken over the past 30 years and include multi-factor approaches. Several approaches have addressed the allergen itself, by modifying IgE reactivity. Others have been directed to the administration route. The immune system is accessible through the oral cavity and sublingual administration of allergens is a known route of administration. Administration can be accomplished by placing the vaccine formulation under the tongue and letting it remain. there for a short time, for example approximately 2 minutes.

Conventionally, allergy vaccine using the oromuco-salt pathway consists of daily progressive dosing of an allergen solution. In comparison, the therapeutic (cumulative) maintenance doses provided exceeded the maintenance doses of a high-factor subcutaneous treatment. . Oromucosal administration is known not to stimulate any or only minimal side effects, primarily in the form of itchy mouth and sometimes itchy eyes and ears.

WO 04/047794 discloses a fast dispersing solid dosage form for sublingual administration of an allergy vaccine.

A commercial allergy vaccine for sublingual administration in the form of drops comprises an allergen dissolved in a liquid mixture of 50% glycerol and 50% water.

Commercial allergy liquid vaccines for sublingual administration are conventionally administered using a dosing regimen comprising a progressive dosing period, typically from 10 to 20 days of daily dosing, whereby the dose is gradually increased to the level of the dosage. maintenance dose, followed by a maintenance period, in which the patient receives a maintenance dose. Recently, a number of studies have investigated the possibility of progressive dosing by the so-called rapid progressive dosing treatment performed for a few days, a day or even a few hours, in which increasing doses are administered with only a short period of time, for example, an hour or two, passing administration entrecada. However, such rapid progressive dosing treatment has the disadvantage that the occurrence of serious side effects cannot be allowed and, consequently, such subcutaneous administration requires the supervision of an allergist. The object of the present invention is to provide a perfect method for prevention or treatment of allergy in an individual through oromucosal administration.

SUMMARY OF THE INVENTION

This object is achieved by the present invention, which relates to the use of an allergen for the manufacture of a liquid vaccine formulation for allergy prevention or treatment in an individual by oromucosal administration in a dosing regimen not comprising progressive modification, i.e. Treatment may be initiated by administering a maintenance dose, for example using the standard maintenance phase dosing regimen.

The present invention is based on a clinical experiment which, surprisingly, has shown that vaccination for allergy by oromucosal administration using a liquid formulation can be performed without any progressive dosing while maintaining an acceptable level of side effects. This finding is very surprising in that, until now, it was believed that a phase of progressive dosing of some kind was essential to avoid serious side effects.

The possibility of avoiding a progressive dosing phase further provides an oromucosal treatment with a liquid devacin formulation with the advantage that this makes the dosing regimen much simpler. In addition, the simple dosing regimen increases patient compliance with treatment.

DETAILED DESCRIPTION OF THE INVENTION

Allergen

The allergen of the formulation according to the present invention may be any naturally occurring protein that has been reported to induce allergy, i.e. IgE-mediated reactions upon exposure to an individual, examples of naturally occurring allergens. include us in pollen allergens (tree pollen allergens, herbs, weed and weed), insect allergens (inhalant, saliva and poison allergens, por8

eg mite allergens, cockroach allergens and mosquitoes, hymenopteran poison allergens), hair allergens are animal dandruff (eg from dog, cat, horse, rat, mouse, etc.) and food allergens. Important pollen allergens from trees, grasses and herbs are those originating from 5 rivers of the taxonomic orders of Fagales, Oleales, Pinales and platanaceae including, for example, birch (Betula), oak (Alnus), walnut (Corylus), carpino ( Carpinus) and olive (Olea), cedar (Cryptomeria and Juniperus), plane tree (Platanus), of the order of Poales including, for example, grasses of the Lolium, Phleum, Poa, Cynodon, Dac.ty.lis, Holcus, Phalaris, Secale and Sor-10 ghum, the orders of Asterales and Urticales including, for example, herbs of the genera Ambrosia, Artemisia and Parietaria. Other important inhalation allergens are those of house dust mites of the genus Dermato-phagoides and Euroglyphus, storage mites, eg Lepido-glyphys, Glycyphagus and Tyrophagus, those of cockroaches, mosquitoes and mos-15cas, for example. , Blatella, Periplaheta, Chironomus and Ctenocepphalides and those of mammals, such as cat, dog and horse, poison allergens, including those originating from insect bites or stings, such as those of the taxonomic order of Hymenoptera, including bees (superfamily Apidae) , wasps (Vespidea superfamily) and ants (For-20 micoidae superfamily). Important inhalation allergens from fungi and molds are, for example, those originating from the genera Alternaria and Cladosporium.

In a particular embodiment of the invention, the allergen is Bet ν

1, Aln g 1, Color a 1 and Car b 1, That a 1, Cry j 1, Cry j 2, Cup a 1, Cup s 1, Jun 1, Jun 2, Jun 3, Ole and 1, Lig ν 1, Pla I 1, Pla a 2, Amb a 1, Amb a 2, Amb t 5, Art ν 1, Art ν 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 ρ 1, Hol I 1, Lol ρ 1 and 5, Pha a 1, Pas η 1, Phl ρ 1, Phl ρ 6, Pol ρ 1, Poa ρ 5, Sec c 1, Sec c 5, Sor h 1, Derf 1, Derf 2, Der ρ 1, Der ρ 2, Der m 1, Eur m 2, Gly d 1, Lep d 2 , Blo 11, Tyr ρ

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 c2, Equ c 3, Mus m 1, Rat η 1, Apis m 1, Api m 2, Ves ν 1, Ves ν 2, Ves ν 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 andSol 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 Alternative Molecular Crossover hybrids of any one of these.

In a preferred embodiment of the invention, the allergen is selected from the group consisting of a tree pollen allergen, a grass pollen allergen, a house dust mite allergen, a weed allergen, a mold allergen. , a cat allergen and a dog allergen.

In yet another embodiment of the invention, the formulation comprises at least two different types of allergens originating from the same allergic source or originating from different allergic sources, for example, group 1 or group 5 gram allergens or group 5 allergens. group 1 or group 2 mite from different grass species and mite, respectively, weed antigens such as small and giant Iosnabranca allergens, allergens from different fungi such as alteraria and cladosporium, allergens such as birch, walnut, car-pin and oak, food allergens such as peanut, soy and milk allergens.

The allergen incorporated in the formulation may be in the form of an extract, a purified allergen, a modified allergen, a recombinant allergen or a recombinant allergen mutant. An allergenic extract may naturally contain one or more isoforms of the same allergen, whereas a recombinant allergen typically represents only an isoform of an allergen. In a preferred embodiment, the allergen is a recombinant allergen. In another preferred embodiment, the allergen is a naturally occurring low binding IgE mutant or a recombinant low binding IgE mutant. In addition, the allergen may be a mixture of a range of allergens, for example from 2 to 10 allergens, in particular from 3 to 9 allergens, more particularly from 4 to 8, and even more particularly from 5 to 7 allergens.

Allergens may be present in equimolar amounts or the proportion of allergens present may vary, preferably up to 1:20. In another embodiment of the invention, the low IgE binding allergen is an allergen according to WO 99/47680 , WO 02 / 40676or WO 03/096869 A2.

The preferred strength of a single dose formulation intended for daily administration is from 1 to 2000 STU, more preferably from 50 to 1000 STU1, more preferably from 100 to 500 STU, more preferably from 130 to 350STU, and most preferably from 150 to 150 STU. at 250 STU.

The preferred strength of a daily dose monodose formulation is from 100 to 500000 SQ-u, more preferably from 200 to 300000 SQ-u, more preferably from 500 to 200000 SQ-u, more preferably from 1000 to 100000 SQ. more preferably from 1500 to 80000 SQ-u and even more preferably from 2000 to 50000 SQ-u.

The amount of allergen which corresponds to a certain power level varies greatly depending on the allergen species. In another embodiment of the invention, the concentration of the major allergen in a single dose intended for daily administration is 0.05 to 50 μg, more preferably 0.05 to 30 μg, more preferably 0.06 to 25 μg, more preferably 0 , 07 to 20 μg, more preferably from 0.08 μg to 15 μg, more preferably from 0.09 μg to 10 μg and even more preferably from 0.1 μg to 7 μg.

Oromucosal Administration

The immune system is accessible through the oral cavity and sublingual administration of allergens is a known route of administration. Administration may be by placing the vaccine formulation under the tongue and letting it stay there for a short time, for example, from 30 to 300 seconds, preferably from 45 to 240 seconds, more preferably from 60 to 180 seconds, more preferably from 90 to 150 seconds, and most preferably from 90 to 120 seconds.

Oromucosal administration comprises any method of administration wherein the formulation, in whole or in part, contacts the oral cavity mucosa and / or the patient's pharynx. Oromucosal administration methods include sublingual administration and buccal administration.

In one embodiment of the invention, the subject is subjected to a vaccination protocol comprising daily administration of the vaccine. In another embodiment of the invention, the vaccination protocol comprises administering the vaccine every two days, every three days or every four days. For example, the vaccination protocol comprises administering the vaccinator for 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 even more preferably more than 36 weeks. weeks

The administration period may be a continuous period. Alternatively, the administration period is a discontinuous period interrupted by one or more non-administration periods. Preferably, the (total) non-administration period is shorter than the (total) period of administration.

In another embodiment of the invention, the vaccine is administered to the patient once daily. Alternatively, the vaccine is administered to the patient twice daily. The vaccine may be a single dose vaccine.

Liquid vaccine formulation

The liquid vaccine formulation of the invention may further comprise any adjuvant and other suitable excipients for such deformulation. Such excipients are well known to those skilled in the art and include, for example, solvents, emulsifiers, wetting agents, plasticizers, coloring substances, fillers, preservatives, viscosity adjusting agents, buffering agents, adjusting agents pH, isotonicity adjusting agents, mucoadhesive substances and the like. Examples of formulation strategies are well known to those skilled in the art.

The adjuvant may be any conventional adjuvant, including oxygen-containing metal salts, for example aluminum hydroxide, heat-sensitive enterotoxin (LT), cholera toxin (CT), cholera detoxin B (CTB), Polymerized liposomes, mutant toxins, for example LTK63 and LTR72, microcapsules, interleukins (e.g. IL-1β, IL-2, IL-7, IL-12, INFy), GM-CSF, MDF derivatives, oligonucleotides deCpG, LPS, MPL, Phosphates, Adju-Phos®, Glycan, Antigen Formulation, Liposomes, DDE, DHEA, DMPC, DMPG1 DOC / Alum Complex, Freund Incomplete Adjuvant, ISCOMs®, Oral LT Adjuvant, Demuramyl Dipeptide monophosphoryl lipid A, muramyl tripeptide and phosphatidylethanolamine.

Aqueous solutions of oxygen-containing metal salts are typically in the form of gels. The concentration of aluminum hydroxide in the formulation is preferably 0.035-1000 mg / ml, more preferably 0.10-100 mg / ml, more preferably 0.25-10 mg / ml and most preferably 0, 5-5 mg / ml. However, the formulation of the invention may also have the form of a highly concentrated gel or a gel-like formulation.

In a particular embodiment of the invention, the formulation according to the invention further comprises an enhancer. An enhancer is a substance which increases the bioavailability of a pharmaceutically active substance (i) by increasing the penetration of the active substance through the biomembrane by opening the paracellular pathways (hermetic junctions), ie. , increased absorption of the active substance and / or ii) through the bioadhesive effect of the enhancer, which increases the absorption period.

Examples of enhancers are an alcohol, for example ethanol, chitosan, chitin, propylene glycol, glycerol, dimethyl sulfoxide, dimethyl formamide, dimethyl acetamide, preferably glycerol. The enhancer is preferably present in an amount of from 20% to 80% (v / v), more preferably from 30% (v / v) to 70% (v / v) and most preferably from 40% to 80% (v / v). % (v / v) to 60% (v / v).

The formulation of the present invention may be prepared by dissolving adjuvants and other excipients, if present in a solvent, preferably water, adding the allergen and allowing the adjuvant allergen and other excipients to react over a period of time. The reaction period may be from 0.1 to 48 hours, preferably from 12 to 24 hours. The reaction is preferably carried out at a temperature of 4 to 45 ° C, more preferably from 4 to 20 ° C. .

Oxygen-containing metal salts

In a preferred embodiment of the invention, the liquid vaccine formulation of the invention comprises an oxygen-containing metal salt adjuvant. Preferably, the metal cation of the oxygen containing metal salt is selected from the group consisting of Al, K, Ca, Mg, Zn, Ba, Na, Li, B, Be, Fe, Si, Co, Cu, Ni. , Ag, Au and Cr.

The anion of the oxygen containing compound can be any oxygen containing ion, including an organic or inorganic anion or a combination of organic and inorganic anions. Examples of metal-containing oxygen salts are, for example, those in which the anion is selected from the group consisting of sulfates, hydroxides, phosphates, nitrates, iodates, bromates, carbonates, hydrates, acetates, citrates, oxalates and tartrates, as well as salts. as mixed forms thereof. Oxygen-containing metal salts further comprise coordination complexes. A definition of coordination complexes is provided, for example, in The Handbook of Chemistry and Physics 56 Ed., Section B, Chapter 7 (197576).

Within the present invention, the term "mixed forms" is intended to include combinations of the various anions as well as combinations, for example with chlorides and sulfides.

Examples of oxygen-containing metal salts according to the invention are aluminum hydroxide, aluminum phosphate, aluminum sulfate, aluminum acetate, aluminum potassium sulfate, calcium phosphate, calcium tartrate, Maalox (mixture of aluminum and magnesium hydroxide), beryllium hydroxide, zinc hydroxide, zinc carbonate, dezinc sulfate and barium sulfate.

More preferred are aluminum hydroxide, aluminum phosphate, aluminum acetate, calcium phosphate, calcium tartrate and zinc sulfate.

The oxygen-containing metal salt pi is typically in the range of 2-11. The pi for allergen proteins is typically in the range of 4-9. Preferably, the allergen and oxygen-containing metal salt are selected such that the allergen's pi is lower than the oxygen-containing metal salt's pi.

When using, for example, aluminum hydroxide as oxygen containing metal salt, the concentration of aluminum hydroxide in the formulation is preferably 0.035-1000 mg / ml, more preferably 0.1 Ο-100 mg / ml, more preferably 0%. 25-10 mg / ml and even more preferably 0.5-5 mg / ml. For the other oxygen containing metal salts, the concentration of the metal salt is preferably 0.035-1000 mg / ml, more preferably 0.35-100 mg / ml, more preferably 0.7-50 mg / ml. even more preferably 1.0-20 mg / ml. The concentration of allergens in the formulation is preferably 0.01-100 mg / ml, more preferably 0.1-10 mg / ml. The proportion of oxygen-containing metal salt for allergen is preferably from 0.1 to 100, more preferably from 1 to 20. The degree of allergen adsorbed to the oxygen-containing metal salt is typically 5 to 99%. more preferably from 10 to 99% of the amount added. Oxygen-containing metal allergen adsorption depends on the buffer system and reaction conditions, including reaction temperature and time, under which adsorption occurs.

Oxygen-containing metal salts may be characterized by a variety of physicochemical parameters, such as adsorption, solubility and dissolution properties, tonic charge measured as the iso-isometric point pi (pH where the net charge of the substance is zero to one. dissociation compound), dissociation constants, complex coordination, electronic configurations, valence, binding and anti-binding orbitals, deposition properties, adhesion properties, surface characteristics, particle characteristics and adjuvant capacity.

The biologically active substance is believed to be adsorbed (or coupled) to the oxygen-containing metal salt and this adsorption contributes to vaccine efficacy. Several factors may be important or influence the adsorption between the active substance and the oxygen-containing metal salt (see, for example, M.. M. Callahan et al., Pharmaceutical Resear-ch Vol. 8, No. 7, 851- 858 (1991) and Vaccine Design, The Subunit and Adjuvant Approach). These factors include pH, the length of time adsorption sandblocking is performed, mixing conditions, concentrations of the various components in vaccines, containers, temperature, storage, buffer and excipients. It has further been found that the adsorption of the substance can be influenced by the net / global charge of the metal salt and the charge of the active substance, both of which are pH dependent. Another feature believed to be of importance is the solubility of the oxygen-containing demetal salts.

Another feature of salts containing oxygen is the protection of the active substance by maintaining the optimal pH for the active substance in the microenvironment, thus preventing acid degradation, or by protecting the active substance against enzymatic degradation, thereby allowing the substance to be. distributed.

In addition, some of the oxygen containing metal salts have a buffering capacity. This can result in an in vivo microenvironment within the vaccine formulation, which protects the active substance from the degradable environment.

Another feature of oxygen-containing metal salt (s) is their ability to adhere to the mucosal membrane. This is commonly believed to be the absorption of the allergen through the mucosal membrane.

For several of the oxygen containing metal salts (eg AI (OH) 3, AIPO4, Ca3PO4), the particle size range is between 0.5 and 15μm.

DEFINITIONS

The term "oromucosal administration" refers to a route of administration where the dosage form is placed under the tongue or anywhere in the oral cavity to allow the active ingredient to contact the oral cavity mucosa or the oral cavity. pharynx to obtain a local or systemic effect of the active ingredient. An example of an oromucosal administration route is sublingual administration.

The term "sublingual administration" refers to a route of administration where a dosage form is placed below the tongue to obtain a local or systemic effect of the active ingredient.

The term "no progressive dosing" means that full doses are used from the first administration onwards, where "full dose" means the dose used for continuous treatment.

The term "liquid vaccine formulation" means any liquid or fluid formulation, including low viscosity solutions, suspensions and gels.

The term "SQ-u" stands for Standardized Quality Unit: ASQ-u is determined according to the ALK-Abelló S / A "SQ biopower" standardization method, where 100,000 SQ units is equal to the standard subcutaneous maintenance dose. . Normally, 1 mg of extract contains between 100,000 and 1,000,000 SQ-Units, depending on the allergen source from which it originates and the manufacturing process used. The precise amount of allergen can be determined by immunoassay, ie the total content of the main allergen and the total activity of the allergen.

The term "treatment" means to cure, alleviate symptoms or inhibit causes of symptoms, in whole or in part.

The term "prevention" means any kind of prophylactic treatment.

In the field of allergy extracts, there is no accepted international standardization method. There are a number of different extract resistance units, ie biopotency. The method employed and the units used usually measure allergen content and biological activity. Examples of these are SQ-Units (Standard Quality Units), BAU (Biological Allergen Units), BU (Biological Units), UM (Mass Units), IU (International Units) and IR (Reactivity Index). ). Accordingly, if extracts from sources other than those disclosed herein are used, they need to be standardized against the extracts disclosed herein in order to determine their potency in SQ units or any of the units mentioned above. The subject matter is addressed in Allergenic extracts, H. Ipsen et al., Chapter 20 in Allergy, Principle and Practice (Ed. S. Manning) 1993, Mosby-Year Book, St. Louis and Lowenstein H. (1980) Arb Paul Ehrlich Inst 75: 122.

The biopotency, that is, the in vivo allergenic activity of a given extract depends on a number of factors, the most important being the content of the main allergens in the extract, which varies with the material composition of the biological source.

The amount of allergen extract in grams to be used to achieve a desired biopotency varies with the type of extract in question and, for a particular type of extract, the amount of dealergen extract varies from batch to batch, with actual biopotency of the extract.

For a given extract batch, the amount of dealergen extract, in grams, to be used to obtain a desired biopotency can be determined using the following procedure:

(a) The biopotency of various amounts of a reference extract is determined using one or more in vivo immunoassays to establish a relationship between biopotency and the amount of reference extract. Examples of such in vivo immunological tests are Skin Sting Test (SPT), Conjunctival Provocation Test (CPT), Allergic Bronchial Stimulation (BCA) and various clinical trials in which one or more allergy symptoms are monitored, see for example Haugaard et al., J Allergy Clinmmunol, Vol. 91, No. 3, pages 709-722, March 1993.

b) Based on the relationship established between the biopotency and the reference extract, the biopotency of one or more doses relevant for use in dosage forms of the invention is selected with due consideration for a balance of factors i) the treatment effect or relieving allergy symptoms, ii) side effects recorded in immunological tests in vivo and iii) the variability of i) and ii) from one individual to another. Equilibrium is made to obtain a maximum optimal therapeutic effect without experiencing an unacceptable level of side effect. Balancing factors is well known to those skilled in the art.

The biopotency of one or more relevant doses found may be expressed in any available biopotency unit, such as SQ, BAU, IR units, cf. above.

(c) From the reference extract, one or more standard biopower reference extracts are prepared and, if used, the biopower unit values of the standard reference extracts are calculated on the basis of the unit biopower value allocated to one or more relevant doses. For example, such a standard for BAU may be obtained from the FDA, as illustrated below.

d) For the standard reference extracts of each type of extract, a series of parameters for evaluation of the biopotency of extracts were selected. Examples of such endpoints are total allergenic activity, the amount of major defined allergens and the overall molecular composition of the extract. Total allergenic activity can be measured using a competitive in vitro immunoassay, such as ELISA and the MagicLite® luminescence immunoassay (LIA), using a stimulated anti-standardized anti-extract mixture obtained using standard methods, eg mouse stimulated antibodies. or rabbit or a serology reservoir for allergic patients. The content of the major allergens can, for example, be quantified by projectile immunophoresis (RIE) and compared with reference standards. The global molecular composition can be examined using, for example, cross immunoelectrophoresis (CIE) and polyacrylamide / sodium dodecyl sulfate gel electrophoresis (SDS-PAGE).

(e) For a given batch of unknown biopotency extract (test extract), the amount of extract to be used to achieve a desired biopotency level (effective dose for use in solid fingerprinting in accordance with this invention) can be determined as follows: for each selected assessment parameter, the test extract is compared to the standard reference extracts using the relevant measurement methods as described above and, based on the measurement results, the amount of extract having the desired biopotency is calculated.SQ-Unit: The SQ-Unit is determined according to the ALK-Abelló S / A "SQ biopower" standardization method, where 100,000 SQ units is equal to the standard subcutaneous maintenance dose. . 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 manufacturing process used. The precise amount of allergen can be determined by immunoassay, ie the total content of the main allergen and the total activity of the allergen.

STU stands for Specific Treatment Unit and STU is an empirical unit based on the biological activity of the respective allergen extract used for allergen-specific allergy vaccination. 1000 STU correspond to the potency of the highest maintenance dose used in a liquid formulation for SLIT, where the maintenance dose is the recommended dose in allergy-specific vaccination to achieve the best safety / efficacy ratio in the average allergic patient population. .

BAU (Biological Allergen Units) is the biological power unit as determined in accordance with FDA requirements for allergen product described in "Quantitative determination of relative potency of allergenic extracts" "ELISA competition assay." Page 15, # 49N-0012, FDA1 October 1993). A dose of 100,000 SQ-Units containing gram extract is equal to a 2600-4700 BAU content according to the above method. Similarly, other extracts may be evaluated according to the above method.

EXAMPLES

Example 1: Sublingual Immunotherapy (SLIT) (SLITone®) without Progressive Dosing - A New Treatment Schedule

Introduction

During the last years, several studies have been performed with sublingual immunotherapy (SLIT), providing evidence of efficacy combined with a favorable safety profile (1). In order for patients to achieve a high level of effect from self-administered treatment such as SLITone, acceptance should be high (2). Given that one of the cornerstones for acceptance is convenience, it is relevant to improve this product characteristic. The excellent tolerance of SLIT has led to the possibility of eliminating the phase-out phase. This would improve patient convenience and may thus increase acceptance and potentially increase the effect of treatment. This study was designed to assess the tolerability of SLITone® without progressive dosing in patients with dust mite or gram allergy.

Material and methods

Medication

SLITone® (ALK-Abelló) is a liquid single-dose formulation containing 0.2 ml solution containing an allergen dose of 200 STU, where the solution is 50% (v / v) glycerol and 50% (v / v). v) water.

Patients

A total of 135 patients were included (66 patients with allergies to the gram and 69 to the mite), aged 7 to 55 years, all diagnosed with moderate to severe rhinitis or rhinoconjunctivitis with / without asthma symptoms due to sensitivity to mites or grams. The diagnosis was made through a positive skin prick test (prick test, ALK-Abelló, 20 S.A., 100 BU / ml) and / or positive IgE (CAP ≥ class 2 in pharmacy). According to WHO recommendations (3), patients excluded from the experiment, where treatment with immunotherapy was contraindicated, were those sensitive to allergens other than those evaluated in the experiment and those who had previously received immunotherapy.

Study Desian

The study was a randomized, multicenter, double-blind, placebo-controlled, parallel group study. The total duration was 90 days.

The patients were allocated into two groups. Group 1 (N = 69) received active treatment during the study. This included 10 days of progressive dosing (T1), 20 days of maintenance treatment (T2), and two consecutive 30-day periods (T3 and T4) under maintenance treatment. Group 2 (N = 66) received progressive pia-cebo dosing, Τ1, placebo maintenance during T2 and started active treatment at the beginning of T3 and continued active treatment until the end of the study (T4). Data were collected at baseline and at T1-T4. Each patient received a card where any clinical incidents related to each dose of treatment were reported. The investigator noted adverse events in the Case Report Forms.

The study was double blind and non-science was not disrupted at any time until after the study was completed. All patients signed a written consent before starting to participate in the experiment. The study was approved by the Ethics Committees of the participating centers, as well as by the Health Authorities.

Treatment

SLITone® (ALK-Abelló) was administered sublingually and composed of mite extract (Dermatophagoides pteronyssinus or Dermato-phagoides pteronyssinus + Dermatophagoides farinae) or an extract of a gram mix (Dactylis glomerata, Phenolum, Phenolum, Phenolum pratense, Poa pratensis, Secale cereale). Treatment consisted of 10 days of progressive dosing with a multidose vial (starting with 1 drop and increasing to 10 drops) and a maintenance phase with monodose containers. The administration was done once a day. The dose of major allergens administered to each common dose container was 2 μg of major allergen 5 (for grams) and 0.8 / 0.4 μg of Der 1 / Der 2 (for mites).

The appearance, smell and taste of placebo treatment were similar to active treatment.

Statistic

Descriptive statistics was used. The description included the number of patients reporting adverse events (AEs) (Ν), the percentage of patients experiencing AEs (%) and the number of AEs (E).

Results

Of the 135 patients included, 123 completed the study. Patient characteristics are shown in Table 1. The patients' ages ranged from 7 to 55 years, with a mean age of 22 in both treatment groups.

In group 1, 36 patients received grass extract and 33 received mite extract compared with 33 who received grass extract and 33 who received mite extract in group 2. A total of 229EA1S were reported; 157 EA1S (68 and 89 in groups 1 and 2, respectively) were assessed as treatment-related by the investigator and included in the analysis. The most frequently reported AEs (> 5% of patients in any group) related to treatment were oral itching, eye itch, rhinitis, and sublingual edema (Table 2); In total, these accounted for 57.3% of all reported AEs. Oral itching was the most frequently reported treatment-related AE; 16 patients (23.2%) reported 28 oral itching events in group 2. Oral itching was the only AE that was reported by> 10% of patients in any group and, as a total, accounted for 42.7% of all. the events. The total number of SLIT doses administered was 6258 and 6081 in groups 1 and 2, respectively. The oral itch rate per dose administered at a time (T1-T4) is shown in Table 3.

The most frequently reported treatment-related AEs for patients <14 years (children) and for patients> 14 years (adolescents and adults) are shown in Table 4.

A total of 5 patients withdrew because of adverse events. In group 1, three subjects withdrew because of a T1 headache, asthma, rhinitis and pruritus at T1; and sublingual edema in T4, respectively. In group 2, two individuals dropped out due to dyspnea and cough in T2 and tongue and mouth edema in T4, respectively.

The purpose of this parallel, randomized, double-blind, placebo-controlled study was to investigate the tolerability of daily administered SLIT without an initial phase of progressive dosing. The results presented here indicate that SLITone® (grams and mites) is well tolerated and that tolerance is not influenced by the omission of the progressive fingersification phase. In general, the number of patients reporting AE was slightly higher in group 2 (receiving progressive dosing with placebo) compared to group 1 (receiving active progressive dosing) and thus the number of events. However, it should be noted that in group 2, a substantial number of events were reported during placebo treatment (T1 and T2). The most frequently reported AE was oral itch. Oral itching is an expected and well-known side effect of sublingual immunotherapy and has been reported to the same degree or more frequently by other authors (4-7).

The progressive dosing method in SLIT is empirical and tradition-based and is likely to be adapted from the gradual increase in dose required for subcutaneous immunotherapy. Data from previous clinical trials (4; 6) and drug-surveillance studies (7) indicate excellent SLIT tolerance, but not until recently, where the impact of progressive dosing step on tolerability has been brought into discussion. The possible impact of progressive dosing on SLIT is speculative. Assuming that the progressive dosing effect on SLIT is so that patients are able to safely administer the maintenance dose after progressive dosing, the data presented here prove that this hypothesis is wrong. The number of patients in group 2 who reported oral itching in T3 (when active treatment was started as the maintenance dose) was higher than in group 1 in T1, but was similar to the number of patients in group 1 who reported oral itching in T2, At this moment, the supposed effect of the progressive dosing phase on the patient's tolerability justified a smaller number. The oral itch rate per dose administered also indicates that these events were reported equally frequently in group 1 at T2 and group 2 at T3. In addition, when comparing the first 30 days of active treatment in both groups (T1 + T2 for group 1 and T3 for group 2), the cumulative rate of oral itching per dose administered was 0.83% and 0.74 % for group 1 and group 2, respectively. The short duration of this study does not allow any firm conclusions as to whether or not the report of oral itching will decrease over time, however, the relatively few dropouts observed indicate that patients do not consider the intensity and duration of problematic oral itching.

Sublingual edema was reported more frequently in group 2 than in group 1. This type of event was reported in group 2 when active treatment was administered (T3 and T4) and resolved within 1 hour at 3 days. One patient dropped out of the study because of his T4 AE after a minimum of 30 days of active treatment. Sublingual edema was also reported in group 1 at T4.

When comparing oral itching in children and adolescents and adults, it appears from the data that oral itching is reported more frequently in children than in adolescents and adults. The reasons for this are not evident, but it remains a topic for further investigation. However, the number of dropouts among children was not different from that of adolescents and adults. In addition, dropouts among children (N = 2) were not related to oral itching.

In conclusion, the results of this study indicate that SLITo-ne® (mites and grams) is well tolerated and that treatment can be started safely without a progressive dosing phase.

(1) Wilson DR1 Torres LM, Durham SR. Sublingual immunothe-rapy for allergic rhinitis: systematic review and meta-analysis. Allergy 2005; 60 (1): 4-12.

(2) Osterberg L. Blaschke T. Adherence to medication. N EnglJ Med 2005; 353 (5): 487-497.

(3) Bousquet J, Lockey RF, Mailing HJ. WHO Position Paper.Allergen immunotherapy: Therapeutie vaccines for allergic diseases. Allergy1998; 53 (44): 1-42. -

(4) Pajno GB1 Morabito L, Barberio G, Parmiani S. Clinical andimmunologic effects of long-term sublingual immunotherapy on asthmaticchildren sensitized to mites: a double-blind, placebo-controlled study. Allergy2000; 55 (9): 842-849.

(5) Canonica GW, Passalacqua G. Noninjection routes for immunotherapy. J Allergy Clin Immunol 2003; 111 (3): 437-448.

(6) Quirino T, lemoli E, Siciliani E, Parmiani S, Milazzo F. Sublingual versus injective immunotherapy in grass pollen allergic patients: double blind (double dummy) study. Clin Exp Allergy 1996; 26 (11): 1253-1261.

(7) Di Rienzo V, Pagani A, Parmiani S, Passalacqua G, Canoica GW. Post-marketing surveillance study on the safety of sublingual immunotherapy in pediatric patients. Allergy 1999; 54 (10): 1110-1113.

Table 1: Patient Characteristics

<table> table see original document page 26 </column> </row> <table> Table 2: Most frequently reported AEs (> 5% in any group) related to treatment

<table> table see original document page 27 </column> </row> <table>

The number of patients (N) reporting the most frequently reported adverse events (> 5% of patients in any group), the percentage of population in the group (%), the number of events (E), and the reporting time in both treatment groups. Group 1 received active T1-T4 treatment, group 2 received placebo during T1 and T2. The total number of patients is corrected for patients who have repeatedly reported the same type of adverse events.

Table 3. Rate of oral itching per dose administered (%) reporting name.

<table> table see original document page 27 </column> </row> <table>

E: Number of oral itching events

Event rate per dose administered (%) in frequently reported adverse event (oral itching) at the time of reporting.

Table 4. Number of patients with most frequently reported treatment-related adverse events by age

<table> table see original document page 28 </column> </row> <table>

Number of patients with the most frequently reported adverse event related to treatment by age.

Claims (11)

1. Use of an allergen, characterized in that it is for the manufacture of a liquid vaccine formulation for the prevention or treatment of allergy in an individual by oromucosal administration in a dosage regimen not comprising progressive dosing. Use according to claim 1, characterized in that the concentration of the main allergen in a single dose for daily administration is 0.05 to 50 μg), more preferably 0.05 μg to μg, more preferably 0, 06 μg to 25 μg, more preferably from 0.07 μg to 20 μg, more preferably from 0.08 μg to 15 μg, more preferably from 0.09 μg to 10 μg and even more preferably from 0.1 μg at 7 μg. Use according to Claim 1 or 2, characterized in that the allergen is selected from the group consisting of a tree pollen allergen, a grass pollen allergen, a house dust mite allergen, a storage mite allergen. , an herb allergen, a mold allergen, a cat allergen and a decay allergen. Use according to any one of claims 1 to 3, characterized in that the oromucosal administration is sublingual administration. Use according to any one of claims 1 to 4, characterized in that the formulation further comprises an enhancer. Use according to claim 5, characterized in that the enhancer is glycerol. Use according to any one of claims 1 to 6, characterized in that the vaccine further comprises an adjuvant selected from the group of oxygen-containing metal salts. Use according to claim 7, characterized in that the oxygen-containing metal salt is selected from the group consisting of aluminum hydroxide, aluminum phosphate and calcium phosphate. Use according to claim 8, characterized in that the oxygen-containing metal salt is aluminum hydroxide. Use according to claim 9, characterized in that the concentration of oxygen-containing metal salt is 0.035-1000 mg / mL, more preferably 0.10-100 mg / mL, more preferably 0.25-10. mg / mL and even more preferably 0.5-5 mg / mL. 11. A method of preventing or treating an allergy in an individual, characterized in that it comprises oromucosally administering to the individual a liquid vaccine formulation in a fingertip regimen not comprising progressive dosing.