DE2141289A1 - Spray vaccines - Google Patents

Description

The invention relates to human and Veterinary spray vaccines.

In British patents 837465 and 994734 pharmaceutical sprays are described in which a powdery Therapeutic agent is dispersed in a liquid propellant gas. In this formulation form, the therapeutic agent can be passed through the nasopharynx with the help of a nebulizer be aerosolized into the upper airways.

According to the two patents mentioned, the dispersion is made with the aid of a liquid nonionic surfactant formed and maintained.

Surprisingly, it has now been found that lecithin can be used to disperse dry prophylactic agents (antigens) in a liquid propellant gas, which substance is neither liquid nor non-ionic.

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Accordingly, the invention relates to human and veterinary spray vaccines, which are characterized by the fact that dry antigens with the help of lecithin are dispersed in a liquid propellant gas.

Among antigens here are both viral and Understand bacterial antigens. This expression denotes both dead and weakened live viruses and bacteria, as well as toxoids.

Bacteria, mycoplasmata and viruses of various types can be processed into vaccines according to the invention. The following can be named: different strains of influenza, such as A -Axchx, A -Japan, A "-Hong Kong, A" -England, B-Johannesburg, B-Massachussets, B-Netherlands; Parainfluenza strains such as Types 1, 2 and 3, adenovirus strains such as types 3> ^ and 7; Measles virus, poliovirus, tetanus, diphtheria and whooping cough bacteria; Reoviruses, Infectious Bronchitis Viruses, "Bovine viral "diarrhea virus, equine influenza virus, rhinoviruses, Rhxnopneumonxtisvirus, "Newcastle Disease" virus, infectious Laryngotracheitxsvxren, "canine herpes" virus, "feline herpes "virus, MiyagawaneIYes. virus, Panleucopeniavirus, Canine distemper virus, rabies virus, pseudo xabies virus, swine fever virus, Foot and mouth disease virus, vaccinia virus, blue tongue virus, Pasteurella multocida; Cocci how Staphylococcus aureus, Staphylococcus albus, Streptococcus, Diplococcvis pneumoniae; also Escherichia, e.g. Coil, Salmonella, Corynebacterium, Actinobacilli, Haemophilus, Neisseria, Proteus, Pseudomonas and the like.

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The vaccines according to the invention are to be understood as meaning both monovalent and multivalent vaccines.

The antigens to be processed in the vaccines can be processed in the manner known for the respective genus can be obtained.

The viruses are obtained by culturing on incubated eggs or in tissue cultures, where appropriate attenuation previously takes place in such media or in animals. The bacteria are mostly made from artificial ones Culture media received.

Virulent antigenic material can interact with the

known agents such as formaldehyde, δ-propiolactone, ultraviolet radiation and heat treatment.

The antigenic material can, if desired be cleaned by common techniques.

The antigens are to be processed in the dry state to form a vaccine according to the invention. she are subjected to a freeze-drying treatment for this purpose, which is optionally followed by a second drying. However, this is not always necessary and depends on the conditions under which the material is subjected after the freeze-drying treatment is processed. If the absorption of moisture is excluded, post-drying can take place be omitted.

The second drying can be done by placing the antigens in a vacuum over a strong for a few days hygroscopic agents, such as concentrated sulfuric acid,

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_{CaCO, Na SO 2} , silica gel, P 2 O ₅ and the like. The treatment can be shortened by increasing the temperature a little, for example to about 35 to> is 50 ° C.

The spray vaccines according to the invention can obtained by dispersing dry antigenic material in a liquid propellant with the help of lecithin will.

It is useful if the antigen suspension is mixed with licithin before the freeze-drying treatment will. In this way, an intimate mixture is obtained in a particularly simple manner. However, it is also possible to use the antigen Put the material and the licithin together only after the freeze-drying treatment.

The amount of lecithin needed to disperse the

antigenic material is required is usually at least 1 mg / ml vaccine fluid. For greater stability To achieve this, the amount of lecithin is preferably increased to 10 mg / ml. But larger quantities can also be used, e.g., 100 mg / ml or more can be used. The upper limit is determined by the solubility in the liquid propellant gas. For practical purposes the amount of lecithin will be between 1 and 100 mg / ml and preferably between about 10 and 100 mg / ml chosen. Naturally, in order to achieve a satisfactory stability of the suspension, the amount is also antigenic Materials per ml matter. In any case, the required amount of lecithin can be determined in a simple manner. For this purpose, the antigenic material is mixed with the liquid

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Propellant gas mixed in a ratio that provides the required concentration, e.g. 10 mg lecithin per ml Suspension can be added. When a homogeneous suspension is obtained, it is determined how much time elapses before that suspended material has settled. If this time is too short, the stability can be increased by adding another Amount of lecithin can be increased.

As propellant gases, the gases customary in pharmacy, which are liquid at room temperature under pressure are, find application. As such, for example, the following can be mentioned: halogenated hydrocarbons, such as dichlorodifluoromethane, Dichlorotetrafluoroethane, trichloromonofluoromethane, dichloromonofluoromethane, monochlorodifluoromethane, trichlorotrifluoroethane, Difluoroethane, monochlorotrifluoroethane, hydrocarbons, such as butane, isobutane, propane and the like. Or mixtures thereof. Mixtures of Gases and gases liquid under pressure at room temperature can be used.

While the invention is relevant to vaccines in general, it is particularly important to Vaccines that serve to protect the respiratory organs because the Corresponding antigens with a vaccine according to the invention can be guided to the point where the Infection, against which the vaccine is supposed to protect, attacks.

In particular, the invention is of importance for influenza vaccines, because these are the most common Infection of the respiratory system are directed.

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The invention is explained in more detail below with the aid of a few examples.
1. Monovalent vaccines

4000 embryonated eggs were inoculated by the allantoic route with influenza virus strain A ₂ -Hong Kong, which was obtained on eggs and mice according to the formula MK EM _1? E_ MK = monkey kidney (monkey kidney), E = egg (Allantoxsweg), Μ = mouse). After an incubation time of 2 days at 35 ° C., the eggs were cooled to k ° C. (16 hours) and the atlantic fluid was collected. The volume obtained was 30 liters.

The virus was purified with the aid of low-line and high-line centrifugation at 1,300 and 50,000 g, the latter treatment being carried out in a so-called Sharpiess centrifuge at a rate of 1.5 l / hour. The sediment from the high-performance centrifugation was resuspended in an isotonic phosphate buffer with a pH of 8.0. Some impurities were centrifuged off at 1300 g. For inactivation, 0.03. % Wt., And then was 0.02 wt. ^ O

β- propiolactone added. The volume of 5 liters was dialyzed against water for 3 days using 10 times the volume of water. The dialysis fluid was replenished once.

Soy lecithin was added to the dialysate in an amount of k mg / 100 CCA virus (CCA = chicken cell agglutination). The whole was freeze-dried in a Leyboldt freeze-dryer of the GO k type. The resulting dry material was subjected to secondary drying

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PO "in a vacuum, after which the dry substance in

. $ Dichlorodifluoromethane and 60 vol. $ Trichloromonofluoromethane was suspended. The product obtained in this way contained 150 CCA virus per ml. 2. Bivalent vaccine

4,000 embryonated eggs were found on the allantox route vaccinated with influenza virus strain A ^ -Aichi on egg according to Formula E. After an incubation period of 2 days at 35 ° C, the eggs were heated to 4 ° C (16 Hours) and the allantox liquid was obtained. The volume obtained was 30 l. 50000 g, the latter treatment in a so-called Sharpies centrifuge with one execution speed of 1.5 l / hour was carried out, the virus was purified. The sediment from the high power centrifugation was in an isotonic phosphate buffer with a pH of 8.0, which contained 0.01 M citrate, resuspended. Some impurities were centrifuged off at 1300 g. For inactivation, 0.03 wt. ^ And then again 0.02% by weight of propiolactone added. The volume of 5 1 was dialyzed against water for 3 days using 10 times the volume of water. The dialysis fluid was refreshed once.

3000 embryonated eggs were inoculated with influenza virus strain B-Massachussets via the allantox route, the on egg according to formula E. was adjusted.

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After an incubation period of 3 days at 33 ° C., the eggs were cooled and the allantoic fluid was collected. The volume was 22 liters. The virus was purified with the aid of high-performance and low-power centrifugation at 50,000 and 1,300 g, respectively, the former treatment being carried out in a so-called Sharpies centrifuge at a rate of 1.5 l / hour. The sediment from the high power centrifugation was resuspended in an isotonic phosphate buffer with a pH of 8.0. Some impurities were centrifuged off at 1300 g. For inactivation were 0.03 wt. ^ And then again 0.02 wt. Tso added 3-propiolactone. The volume of 3.5 liters was dialyzed against water for 3 days using 10 times the volume of water. The dialysis fluid was replenished once.

After dialysis, the two were obtained

Crowds mixed. The ratio between the amount of strain A virus and the amount of strain B virus was 3: 2.

1 mg of lecithin was added per 100 CCA virus. The whole was freeze-dried in a Leyboldt freeze-drying apparatus of the GO h type, after which the dry substance was suspended in ^ JO vol. ^ Dichlorodifluoromethane and 60 vol. $ Trichloromonofluoromethane. The product thus obtained contained 1500 CCA Ag virus and 1000 CCA B virus per ml.

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Claims (1)

2 U1289 PATENT CLAIM: 1. Human and Veterinary Spray Vaccines, thereby characterized that dry antigens mib help from lecithin are dispersed in a liquid propellant gas, 2. Spx'ay vaccines according to claim 1, characterized in that that antigens are dispersed to prevent infections of the respiratory system. 3. Spray vaccines according to claim 2, characterized in that that influenza antigens are dispersed. k. Spray vaccines according to one of Claims 1 to 3i characterized in that it contains at least 1 mg of lecithin per ml included. 5 · Spray vaccines according to claim h, characterized in that they contain 1 - 100 mg lecithin per ml. 6. Spray vaccines according to claim 5 »characterized in that that they contain 10-1QO mg of lecithin per ml. 7. Process for the production of human and veterinary Vaccines, characterized in that dry antigens be dispersed in a liquid propellant with the help of lecithin. 8. The method according to claim 7 »dadLirch characterized, that antigens are dispersed to prevent infections of the respiratory system. 9. The method according to claim 8, characterized in that that influenza antigens are dispersed. 10. The method according to any one of claims 7 to 9, characterized in that at least 1 mg of lecithin is used per ml of vaccine fluid. 209811/1780 11. The method according to claim 10, characterized in f, v; kenrizeichne t das3 1 - 100 mg lecithin per ml vaccine liquid is used will, 12. The method according to claim 11, characterized in that that 10-100 mg of lecithin is used per ml of vaccine fluid will. 209811/1780