RU2548727C2 - Allergotropines for treatment of allergy caused by house dust mites - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to the field of medicine, namely to immunology and allergology, and can be used for the treatment of allergy to house dust mites. For this purpose an allergotropine is obtained by a method of chemical conjugation of an allergoid and synthetic highly molecular immunomodulator Polyoxidonium. The allergotropine contains 1000±200 PNU of the allergoid of house dust mites Dermatophagoides pteronyssinus or Dermatophagoides farinae and 6.25±1.25 mg of Polyoxidonium.

EFFECT: obtaining the allergotropines, based on the allergoid forms of allergens of house dust mites Dermatophagoides pteronyssinus or Dermatophagoides farinae, possessing more expressed immunogenic and hyposensitising properties in comparison with the native allergen, makes it possible to carry out the effective treatment of allergy by ASIT method.

2 ex, 2 tbl, 2 dwg

Description

The invention relates to medicine, namely to immunology and allergology, and for the treatment of allergies to house dust mites.

At present, allergy-specific immunotherapy (ASIT), for example, pollinosis, is carried out both by the method of subcutaneous injection (SCACIT) and sublingually (slACIT). For injection forms of drugs, water-salt extracts (VCE) of allergens, modified allergens (allergoids), as well as deposited allergens and allergoids are used. For slasit use water-salt extracts of allergens and allergoids.

There are known water-salt extracts of allergens and allergoids for ncACIT, presented on the pharmaceutical market by domestic preparations: “House dust dust allergen for diagnosis and treatment”, registration number 94/161/20, (Biomed named after II Mechnikov, Russia); “Allergen from the tick Dermatophagoides pteronissimus for diagnosis and treatment”, registration number 89/686/16, (Biomed named after II Mechnikov, Russia); “Dermatophagoides farinae tick allergen for diagnosis and treatment”, registration dead 92/203/19 (Biomed named after II Mechnikov, Russia); "Allergoid from house dust for treatment" registration number P N 001536/01 dated 03/05/2009 ("Microgen" NPO FSUE, Stavropol).

VSE allergens with their relative cheapness and availability on the pharmaceutical market do not differ in safety and ease of use. The main disadvantage of FEV is the high content of ballast substances, which are potential allergens, which creates a risk in the treatment - obtaining anaphylactic type reactions.

Allergoids (NPO Microgen) are relatively cheap and available on the pharmaceutical market, are characterized by a reduced allergenicity compared to the HSE allergens and, consequently, greater efficacy and lower risk, treatment takes less time. However, the administration of pACACIT by allergoids does not ensure a uniform intake of the drug into the patient’s body; moreover, the indicated allergoids existing at a given time are often not stable enough.

Deposited allergic vaccines for PCACIT are known, represented by the Alustal preparation “Tick Allergen”, registration number LP-001047, registration date: 10.21.2011, (Stallergen JSC, France). "Alustal" is an allergen adsorbed on a suspension of aluminum hydroxide. If there is such an advantage of the Alustal preparation as the prolonged action of an allergy vaccine injection, one cannot fail to note the main drawback caused by the presence of an allergic vaccine in the composition of the allergy vaccine - the risk of anaphylactic type reactions. In addition, the deposited drugs cause the formation of painful compaction at the injection site.

There are known oral forms of allergy vaccines based on the VSE allergens presented by preparations in the form of solutions: “Staloral” “Tick Allergen”, registration number LSR-008340 / 10-180810 (Stallergen, France) and “N-Al” “Tick mixture” (“ Sevapharma ”, Czech Republic), registration number LSR-003619/09 dated 05/14/2009. Oral forms of allergy vaccines based on allergoids (in the form of tablets) are presented with the preparation “Lys Dermatophagoides”, registration number LP-001303 dated 11/29/2011 (Lofarma SPA, Italy). Slasit preparations are convenient to use, but expensive and not always readily available in the pharmaceutical market. The SLACIT method is convenient to use, but does not provide the possibility of a uniform flow of the drug into the patient's body. In addition, one cannot fail to note the high allergenicity of drugs based on VSE allergens.

Based on the foregoing, it is clear that the main goal to which this invention is directed is to improve allergy vaccines, namely, to reduce allergenicity, while maintaining immunogenicity, increase the stability of allergic vaccines, the creation of prolonged dosage forms.

A known method of producing allergoids by modifying allergens with formaldehyde (V. A. Fradkin "Diagnostic and therapeutic allergens", Moscow, Medicine, 1990).

Thus, the closest analogues for the intended purpose have not been identified.

Known allergotropin (RF patent No. 2205661 "Allergotropin for the treatment of pollinosis and a method of treating pollinosis"). This allergotropin is characterized in that it contains an allergoid of pollen from allergenic plants (birch, timothy, or wormwood) and the associated synthetic immunomodulator, Polyoxidonium. This allergotropin is intended for the treatment of pollinosis caused by pollen of timothy, birch or wormwood and, accordingly, is not an analogue of the invention for its intended purpose. However, the structure of the known allergotropin can be called an analogue of the claimed invention.

Allergotropins are an allergoid conjugate prepared from a purified allergen-specific freeze-dried water-salt extract of the allergen and a synthetic high molecular weight immunomodulator - Polyoxidonium. Polyoxidonium (a copolymer of N-oxide-1,4-ethylene piperazine and N-carboxyethyl-1,4-ethylene piperazinium bromide), registration number 96/302/9, RF patent N 2073031, FSP 42-0501460703, readily soluble in water, non-toxic, breaks down and is completely eliminated from the body.

An allergoid is a chemically modified allergen and has advantages over the latter, because with chemical modification, a decrease in allergenicity and the risk of anaphylactic shock is observed. The conversion of an allergen to an allergoid by chemical modification in the presence of formaldehyde, leading to the interaction of high and low molecular weight components, helps to preserve the properties of the allergoid to induce antibody synthesis. This action is enhanced by the rigid, stable structure of the allergoid molecule, which is resistant to the effects of denaturing agents, in contrast to the initial allergen, and also helps to prolong the action of the allergoid, since its destruction in the patient’s body will be slower.

The decrease in allergenicity in polymerized allergens is associated, on the one hand, with a decrease in the number of antigenic determinants as a result of their reaction with aldehyde groups, and on the other hand, with the inaccessibility of the remaining determinants hidden inside the high molecular weight compound to reagins. An increase in molecular weight and the creation of more rigid intramolecular bonds have a positive effect on immunogenicity.

The active principles of allergotropins are an allergoid with reduced allergenic activity (compared with the initial allergen), and the immunomodulator Polyoxidonium. The drug must contain a specifically active antigen and have the ability to interact with specific IgG antibodies of the control positive blood serum of patients with hypersensitivity to the allergen.

Based on studies, it was found that allergotropins retain their biomedical and physico-chemical properties for 3 years.

Using a combination of an allergoid and an immunomodulator allows you to reduce the course of treatment (from 32 injections during pCACIT VSE allergen to 15), which reduces the possibility of complications (anaphylactic reactions) during the course of treatment.

Thus, the high efficiency, safety and low cost of allergotropins makes them extremely promising and competitive in the market of antiallergic drugs.

The objective of the invention is to improve allergy vaccines, and the technical result is the creation of drugs for specific immunotherapy of allergies to house dust mites.

This problem is solved, and the technical result is achieved due to the fact that allergotropin has been developed for the treatment of allergies caused by house dust mites, characterized in that it is obtained by the chemical conjugation of an allergoid and a synthetic high molecular weight immunomodulator - Polyoxidonium, and containing 1000 ± 200 PNU of the home tick mite allergoid dust of Dermatophagoides pteronyssinus or Dermatophagoides farinae and 6.25 ± 1.25 mg of Polyoxidonium.

Thus, to solve this problem, the Pterpol allergotropins were developed - for the treatment of allergies to ticks Dermatophagoides pteronyssinus and "Farpol" - for the treatment of allergies to ticks Dermatophagoides farinae. Drugs are obtained by chemical conjugation of an allergoid and Polyoxidonium.

Allergotropins Pterpol and Farpol contain 1000 ± 200 PNU of the allergoid of house dust mites Dermatophagoides pteronyssinus or Dermatophagoides farinae and 6.25 ± 1.25 mg of Polyoxidonium.

The preparations of allergotropins of the invention have a number of obvious advantages, namely: with respect to the VSE of allergens - reduced allergenic activity, enhanced immune response, prolonged action; regarding allergoids, by an enhanced immune response, prolonged action; relatively deposited forms of FEV of allergens - reduced allergenic activity, enhanced immune response, lack of painful compaction at the injection site due to the water-soluble basis of the polymer carrier.

The invention is illustrated by the following graphic materials: table 1 presents the results of determining the immunogenicity of allergotropin "Pterpol", table 2 presents the results of determining the immunogenicity of allergotropin "Pterpol", in FIG. 1 are plotted as the OD values in ELISA for neutralizing IgE antibodies for allergen, allergoid and allergotropin Pterpol, in FIG. 2 are presented in the form of a graph of the OD value in ELISA for neutralizing IgE antibodies for allergen, allergoid and allergotropin “Farpol”.

Allergotropine for the treatment of allergies caused by house dust mites was obtained by chemical conjugation of an allergoid and a synthetic high molecular weight immunomodulator - Polyoxidonium. Allergotropin contained 1000 ± 200 PNU of the allergoid of house dust mites Dermatophagoides pteronyssinus or Dermatophagoides farinae and 6.25 ± 1.25 mg of Polyoxidonium.

The implementation of the invention is illustrated by examples.

Example 1

Allergotropin "Pterpol" is an allergoid conjugate from house dust mites Dermatophagoides pteronyssinus, prepared from a highly purified lyophilized water-salt extract, and a synthetic high molecular weight immunomodulator Polyoxidonium.

One bottle contains 1000 + 200 PNU allergoids from house dust mites and 6.25 + 1.25 mg of Polyoxidonium.

Allergotropine is presented in the form of a lyophilisate for the preparation of a solution for subcutaneous administration. Dilution of the lyophilisate is carried out using a diluent for non-infectious allergens (FSP 42-0010-2041-01).

Allergotropine is used for specific immunotherapy (SIT) of patients with hay fever with sensitization to house dust mites D.pteronysinus.

Allergotropine is prescribed in the presence of an anamnesis and clinical disease with manifestations such as rhinoconjunctivitis and atopic bronchial asthma IA 1; as well as with an established diagnosis of allergy, confirmed by provocative nasal tests and skin tests with a commercial allergen of house dust mites D.pterony sinus.

Allergotropine is stored in a dry, dark place at a temperature of 4 to 8 ° C. Shelf life 3 years.

Since the developed drug is intended for the treatment of allergopathology, conjugation with Polyoxidonium should not change the allergenic properties of the drug in comparison with the initial allergoid.

Immunogenicity and residual allergenicity were compared. Moreover, immunogenicity should not be lower, and residual allergenicity not higher than the corresponding values for the initial allergoid.

Determination of immunogenicity and residual allergenicity of the drug was carried out in an enzyme-linked immunosorbent assay (ELISA). On polystyrene tablets were applied, respectively:

- an allergoid made from the allergen of house dust mites Dermatophagoides pteronyssinus, obtained at the FSBI “SSC Institute of Immunology” of the FMBA of Russia;

- Allergotropin "Pterpol" obtained at FSBI "SSC Institute of Immunology" FMBA of Russia.

The immunogenicity of the drug is determined by its ability to interact with specific IgG from the blood serum of patients. As a positive control, we used pooled serum with a high content of specific IgG antibodies. Serums for it containing at least 10 mgA / 1 were selected using commercial enzyme-linked immunosorbent assay kits on an ImmunoCAP instrument (Pharmacia biotech). The results of determining the immunogenicity of allergotropins are presented in table 1.

It is seen that the optical densities of the obtained preparations and controls are very close, the difference is not significant (p> 0.05). Thus, it was shown that the immunogenicity of the Pterpol allergotropine we made did not change compared to the initial allergoid.

Like the initial allergoid, the manufactured drug should have reduced specific activity compared to the purified allergen from which the drug was prepared - the allergenic activity of the original allergen should exceed the allergenic activity of the allergoid or allergotropin by at least 40%.

The determination was carried out by ELISA inhibition. As a positive control, a pooled serum with a high content of specific IgE antibodies was used. Serums for it containing at least 85 kUA / 1 were selected using commercial enzyme-linked immunosorbent assay kits on an ImmunoCAP instrument (Pharmacia biotech).

The results are presented in FIG. 1. The reaction results were evaluated on the graph by the difference in the OD of the compared samples, at two points in the linear portion of the calibration curves, the difference should be at least 40%. From FIG. 1 shows that the binding of the allergen to the specific IgE of the control serum is higher, because the values of OD in ELISA when neutralizing antibodies are significantly lower than for the corresponding concentrations of the allergoid and the drug, which indicates better binding to specific IgE of the control serum. At two points of the linear portion of the calibration curve, the difference in OD for the allergen and allergoid, together with the preparation based on it, exceeded 40%. Thus, the Pterpol allergotropin, like the initial allergoid, is less allergenic than the house dust mite allergen Dermatophagoides pteronyssinus.

Example 2

Allergotropin Farpol is an allergoid conjugate from house dust mites Dermatophagoides farinae made from highly purified lyophilized water-salt extract and a synthetic high molecular weight immunomodulator Polyoxidonium.

One bottle contains 1000 + 200 PNU of the allergoid from house dust mites Dermatophagoides farinae and 6.25 + 1.25 mg of Polyoxidonium.

Allergotropine is presented in the form of a lyophilisate for the preparation of a solution for subcutaneous administration. Dilution of the lyophilisate is carried out using a diluent for non-infectious allergens (FSP 42-0010-2041-01).

Allergotropine is used for specific immunotherapy (SIT) of patients with hay fever with sensitization to ticks of house dust Dermatophagoides farinae.

Allergotropine is prescribed in the presence of an anamnesis and clinical disease with manifestations such as rhinoconjunctivitis and atopic bronchial asthma IA 1; as well as with an established diagnosis of allergies, confirmed by provocative nasal tests and skin tests with the commercial allergen of house dust mites Dermatophagoides farinae.

Allergotropine is stored in a dry, dark place at a temperature of 4 to 8 ° C. Shelf life 3 years.

Since the developed drug is intended for the treatment of allergopathology, conjugation with Polyoxidonium should not change the allergenic properties of the drug in comparison with the initial allergoid.

Immunogenicity and residual allergenicity were compared. Moreover, immunogenicity should not be lower, and residual allergenicity not higher than the corresponding values for the initial allergoid.

Determination of immunogenicity and residual allergenicity of the drug was carried out in an enzyme-linked immunosorbent assay (ELISA). On polystyrene tablets were applied, respectively:

- an allergoid made from the allergen of house dust mites Dermatophagoides farinae, obtained at the FSBI “SSC Institute of Immunology” of the FMBA of Russia;

- Allergotropin “Farpol”, obtained at the FSBI “SSC Institute of Immunology” FMBA of Russia.

The immunogenicity of the drug is determined by its ability to interact with specific IgG from the blood serum of patients. As a positive control, we used pooled serum with a high content of specific IgG antibodies. Serums for it containing at least 10 mgA / 1 were selected using commercial enzyme-linked immunosorbent assay kits on an ImmunoCAP instrument (Pharmacia biotech). The results of determining the immunogenicity of allergotropins are presented in table 2.

It is seen that the optical densities of the obtained preparations and controls are very close, the difference is not significant (p> 0.05). Thus, it was shown that the immunogenicity of the Farpol allergotropine manufactured by us has not changed in comparison with the initial allergoid.

Like the initial allergoid, the manufactured preparation should have a reduced specific activity compared to the purified allergen from which the preparation is prepared, the allergenic activity of the initial allergen should exceed the allergenic activity of the allergoid or allergotropin by at least 40%.

The determination was carried out by ELISA inhibition. As a positive control, a pooled serum with a high content of specific IgE antibodies was used. Serums for it containing at least 85 kUA / 1 were selected using commercial enzyme-linked immunosorbent assay kits on an ImmunoCAP instrument (Pharmacia biotech).

The results are presented in FIG. 2. The reaction results were evaluated on the graph by the difference in the OD of the compared samples, at two points in the linear portion of the calibration curves, the difference should be at least 40%. From FIG. 2 shows that the binding of the allergen to the specific IgE of the control serum is higher, because the values of OD in ELISA when neutralizing antibodies are significantly lower than for the corresponding concentrations of the allergoid and the drug, which indicates better binding to specific IgE of the control serum. At two points of the linear portion of the calibration curve, the difference in OD for the allergen and allergoid, together with the preparation based on it, exceeded 40%. Thus, the Farpol allergotropin, like the initial allergoid, is less allergenic than the allergens of house dust mites Dermatophagoides farinae.

Table 1 Determination of the immunogenicity of allergotropin "Pterpol" Drug form Manufacturer IgG antibodies (optical density) M ± m Number of experiments Allergoid Institute of Immunology 1.322 ± 0.137 5 Pterpol Institute of Immunology 1.418 ± 0.164 5

table 2 Determination of the immunogenicity of allergotropin "Farpol" Drug form Manufacturer IgG antibodies (optical density) M ± m Number of experiments Allergoid Institute of Immunology 1.291 ± 0.138 5 Farpol Institute of Immunology 1.309 ± 0.143 5

Claims (1)

Allergotropine for the treatment of allergies caused by house dust mites, characterized in that it is obtained by chemical conjugation of an allergoid and a synthetic high molecular weight immunomodulator - Polyoxidonium, and containing 1000 ± 200 PNU allergoids of house dust mites Dermatophagoides pteronyssinus or Dermatophagoides farinae and 6.25 ± 5.25 mg Polyoxidonium.

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