DD213351A1 - METHOD FOR THE PRODUCTION OF NON-PARENTERAL INACTIVE VACCINES AGAINST VIRUSES - Google Patents

Abstract

The method of producing non-viral inactive vaccines against viruses and in combiners therewith against pathogenic microorganisms is applicable in technical virology, microbiology and biochemistry, in particular with regard to the production of vaccines, interferon inducers and special biopraparates. The goal i. the production of inactive vaccines against viruses and in connection with this vaccine in the further other microbially produced inactive vaccines. After non-parenteral administration of these inactive vaccines, a combined utilization of d. antigenic / immunogenic effect u. an interferon induction can be achieved. The task z. Production of high-quality nonparenteral inactive vaccines is thereby achieved in such a way that viruses, or in combination with these pathogenic microorganisms, are targeted in their sensitive center in such a way that a gentle, but complete loss of infectivity occurs, preserving the integrity of the corresponding germs while substantially retaining the native structure of the antigen proteins and the Interferoninduktoren be particularly spared. This targeted inactivation is carried out by gamma rays, microwaves, thermal manipulations or coupled physicochemical methods with a specifically set concentration and, if appropriate, degree of purification of the viruses or microorganisms. The areas of application are the production of inactive vaccines and the preparation of other biopraparates for human and veterinary medicine.

Description

Title of the invention

Process for the preparation of non-parent Inaktivijspfstoffen against viruses

Field of application of the invention

The invention relates to the tec h African production of Inaktivimpfstoffen against viruses for non-parenteral administration and also includes special microbial Inaktivimpfstoffe "These vaccines can be technically simple with few steps and with low yields produce" They allow the production of vaccines with high antigenic / immunogenic properties, which cause a good immunogenic effect and an interferon induction even in the case of nonparenteral administration

The field of application relates to manufacturing plants and other institutions that produce vaccines, interferon inducers or other special preparations.

Characteristic of the known technical solutions

Sine oral immunization with live virus has been described repeatedly. When using live vaccines, however, relatively often unwanted Kebenreaktionen occur. This also applies equally to the use of recombinants, attenuated, sugar-coated, passionate or modified live viruses of the influenza types or subtypes.

Inactivated viruses as orally administered vaccines previously had to be given in very high concentrations to cause significant antibody conversions in respiratory secretions.

The causes can be seen in the chemical inactivation with, for example, formaldehyde or ß-propiolactone, which lead to irreversible reactions with the determinant groups of the antigens and with the nucleic acids · The structural changes of the nucleic acids reduce i.a. also their effect as interferon inducers. Thus, vaccinations with inactivated, non-viable non-parent viruses have so far not prevailed in practice »

The production of highly effective non-parent inactive vaccines has not yet been made on an industrial scale.

Object of the invention

The object of the invention is to develop a method by which it is possible to produce inactive vaccines against viruses and in connection with these vaccines further other inactive vaccines prepared on a microbial basis. After application of the inactive vaccines (inactivated viruses or in combination with inactivated microorganisms), a combined utilization of the antigenic / immunogenic effect and an interferon induction should be achieved.

The useful effect is as follows: Compared with conventional parenteral vaccines to be administered in full virus vaccines, the more or less carried out high-grade purification and, in addition to cleavage and subunit vaccines, there is no need for cleavage by chemicals or enzymes. Moreover, as in the case of adsorbate vaccines, the use of adsorbers / adjuvants (e.g., aluminum compounds) may also be unnecessary. With this procedure should simultaneously; be achieved that viral and

microbial vaccines can be combined as desired and the vaccines can be administered simply (non-parenterally). "Furthermore, it is intended to select such inactivating conditions which allow the integrity of the corresponding germs while largely retaining the native structure of the corresponding antigenic proteins ,

Explanation of the essence of the invention

The invention has for its object to provide a method for producing inactive vaccines against viruses and in combination with it against pathogenic microorganisms, so that due to the special preparation and form of these vaccines, a preferred non-parenteral administration can be carried out.

The object is achieved according to the invention in that viruses or in combination with these pathogenic microorganisms are targeted in their sensitive center in such a way that a gentle but complete loss of infectivity occurs and the decisive antigenic proteins and interferon inductors are particularly protected. " For example, by gamma rays, microwaves or corresponding thermal manipulations or by coupled physico-chemical process steps. The egg. nigungs- and concentration level can be set variable.

The order of the main process steps can be represented as follows, for example:

1. Virus multiplication

2. Low viral enrichment (ultracentrifugation or ultrafiltration)

3. Inactivation (ionizing radiation and / or thermal treatment up to maz »25 ⁰ C)

4. Final preparation as a vaccine for non-parenteral administration

embodiments

example 1

Influenza viruses (strain A / Brazil / 11/78) are propagated after the standard period in fertile chicken eggs. The virus-containing allantoic fluid is centrifuged to max. 10 000 g iron cells and egg components are released. The further low concentration is carried out by ultrafiltration with a commercial apparatus and a cellulose acetate membrane with a selectivity of 87.5 % (compared to dextran 110 000). The filtration rate is at a pressure of about 0.2 MPa 10-50 lm. The filtrate contains no virus or its constituents Concentrated on an enrichment of the virus antigens with 50-80 μg hemagglutinin / ml Yirus inactivation is as follows: Radiation system RCH Gamma-30, Uuklid cobalt-60, total dose 15,000 Gray, T 15 ⁰ G · - Hemagglutinin content and hayaminidase activity hardly or not decrease under these specific conditions, and the integrity of the yiren is also maintained, which has a positive effect on the dosage form (nonparenteral) depending on the type and concentration of the vaccine used. _... -

Example 2

Virus-containing allantoic fluid (strain B / Singapore / 222/79) is kept at about 37 ⁰ C for 2 to 5 days. After detection of the non-proliferative capacity (if necessary prolongation of the storage period), a fur-setting takes place at about 55 000 g. Regarding the application is proceeded as in Example 1 ..

Example 3

A virus suspension (influenza virus type A - hekombinante UIB-4) with a deragglutinin content of about 60 / μg / ml obtained by pelleting according to example 2 is exposed to microwave energy in an amount of 20 ml in a sealed glass vessel. For this serves a commercial Hikrowellengerät (eg Philips DX 260, Litton Industries model 402 002) · The irradiation time lies between? up to 14 seconds (frequency 2450 MHz ± 20 MHz, power output 1.5 kW). - Sine certain cooling during irradiation can be done. - Immediately after the irradiation is cooled down (4 ⁰ C). After detection of the non-proliferation of the viruses, non-parenteral administration is carried out according to Example 1.

Claims (8)

invention claim A process for the preparation of non-parent inactive Yira vaccines, characterized in that such vaccines are produced which exhibit combined utilization of antigenic / immunogenic activity and interferon induction while maintaining the integrity of the viruses, e.g. Influenza viruses, on the fly, allow the virus material to be concentrated or concentrated ^:; 'Unconcentrated or purified or unpurified form is gently inactivated while largely maintaining the native structure of the corresponding antigenic proteins, and in such a way that the sensitive center is gesielt made so that thereby a gentle, but complete loss of infection occurs, especially brought about by Direct or indirect action of radiation, primarily by ionizing radiation in the form of gamma rays in the range of 1 000 to 25 000 Gray, by use of a liquid medium (homogeneous virus suspension) or of Yira material with a HpO content of at least 0.5%, with the use of a dormancy ^% . for the water radical effect and under variable oxygen partial pressures, which leads to a virus inactivation, but does not affect the immunogenicity of the essential viral components. 2. The method according to item 1, characterized in that in the case of liquid media or virus material with water content, the Radiolyseprodukte formed, such as e ~ Η *, · 0Β, HO ₂ ', H ₂ O ₂ , H ₂ , are exploited. 3. The method according to item 1, characterized in that between 0 - 35 ⁰ C or lyophilized or frozen virus material can be inactivated. 4 ·. Method according to item 1, characterized in that the foreign substances from the host cells exert a protective function during the irradiation or a combinatorial action in the vaccine. 5. The method according to item 1, characterized in that also .B irradiations are made with electron beam or ultraviolet light for inactivation, A method according to the first to fifth aspects characterized in that the vaccine is prepared so as to be in liquid or solid form, e.g. Capsule, dragee or tablet or as a spray or aerosol is given non-parenterally for single or "repeated sole immunization or in combination with other ¥ akzinierungen. 7. Method according to item 6, characterized in that, after the sole or combined administration of the inactivated vaccine in man, a load-bearing immunity against the human yirion (s) used to prepare the vaccine, e.g. Influenza strains, detectable. 8. The method according to the items 1 to 5 »characterized in that this procedure can also be applied to microbial inactive vaccines.