DD289200A5 - METHOD FOR PRODUCING A COMBINATION VACCINE WITH INTENSIVE EFFECT - Google Patents

Abstract

The invention relates to the production of a combination vaccine with intensified action and high efficacy against respiratory infections in the human and veterinary medical field. Essentially, the preparation of the combination vaccine relies on a physiological, easily isolatable natural product in association with bacterial components in enteric-coated form. According to the invention, a pressed juice is obtained from raw potatoes, from which lectin is obtained as an immunomodulator by uncomplicated centrifugation and concentration steps. In lyophilized form, this product is converted with inactivated and partially modified bacterial components into an air-, water- and acid-resistant oral vaccine. In contrast to the production of vaccines without lectin, these preparations produce a highly effective preparation. {Bacteria; disintegration; Gamma rays; glycoproteids; Immunomodulators; immunostimulants; inactivation; interferon inducers; potatoes; lectins; lyophilisates; lysates; Enteric Coating; Vaccine}

Description

Field of application of the invention

The application relates to the production of a combination vaccine with intensified action and high efficacy against respiratory infections in the human and veterinary field.

The application is possible for facilities producing vaccines and immunomodulators for human or veterinary use.

Characteristics of the known state of Tochnlk

Methods for producing vaccines against different agents are widely used. This also includes the production of polyvalent vaccines, as described, for example, in DD PS 230775. Similarly, methods for the isolation of lectins are known. This concerns lectins, for example, from plants such as peanut, beak bean, laburnum, gorse, vetch and the like.

However, the specific direction of producing combined vaccines using lectins as an immunomodulator has not been specifically described. This includes both parenteral and nonparenteral vaccines.

Object of the invention

The aim of the invention are those measures from which a method results, which under consideration of application criteria for the production of a combination vaccine to be administered orally with intensified and broad effect against respiratory infections (resistance) leads.

These production measures result in a combination vaccine which, in addition to the formation of specific antibodies, at the same time triggers an interferon induction and a strong activation of the phagocytosis-Klingling systems or activation of intracellular and extracellular enzymes of alveolar macrophages. Essentially, the composition of the vaccine consists of a physiological, easily isolatable natural product in combination with inactivated or modified bacterial components in the form of an enteric final preparation.

Explanation of the essence of the invention

The object, which is solved by the invention, is to provide such measures in addition to microorganisms or components thereof protein or glycoprotein in the form of lectins, which have specific binding sites for carbohydrate structures and react differently with cells, but do not trigger pathological reactions to include as an immunomodulator and to come to a qualitatively novel vaccine with broad specificity against respiratory infections or to a clear activation of defense mechanisms in human mucous membranes. These measures are aimed at the

Simple preparation of the immunomodulator from natural products

- Easy to combine with inactivated or partially split bacterial cell

- Ease of use of the vaccine including prolonged use without unwanted side effects

- intensified effect in this combination over conventional bacterial products

- good storability of the vaccine

The object is achieved in that from raw potatoes, a pressed juice is obtained, which is the biological

active lectin contains STA, whereby after antioxidant addition by uncomplicated centrifugation and concentration steps, a lyophilized immunomodulator is obtained, which is in low concentrations after combination with inactivated and

Part of digested bacterial components and after production of a gastric juice-resistant final form one

optimally effective oral vaccine results.

Surprisingly, it has been found that it is precisely this combination and even small amounts of the immunomodulator and

modified bacterial mixture (= 2-40 ^ g immunomodulator determined in the form of lectin and 2-20mg Lyophilisatbakterieller origin) per dose can induce these high effects against respiratory infections.

The method according to the invention is suitable for the production of optimally acting vaccines for use in human

and veterinary area.

embodiments example 1 Preparation of immunomodulator .STA-R "(crude lectin) from Solanum tuberosum:

- Mechanical comminution of raw potatoes and extraction of the pressed juice by centrifugation. Optionally, add 0.2mg of ascorbic acid to 6ml of pressed juice, spin for 20min at 6000g.

- Testing of the supernatant (lectinhaltig) on its Lectingehalt by agglutination titer against untreated erythrocytes (preferably human, blood group B)

- Lyophilisation (previous deep-freezing to -40 ° C) and re-determination of the lectin content: Determination of the lectin content in μg (using a reference lectin)

Adjustment of STA-R to a lectin content which still agglutinates untreated human B erythrocytes 1: 4 (4HAU) Bacteria or lysates and lyophilisates:

The following bacteria are bred: Escherichia coli, Haemophilus influenzae, Klebsieila pneumoniae, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes for the human area and additionally with Bordetella bronchiseptica, Pasteurella multocida for the

veterinary range inactivation / digestion: Depending on the stability of the germs 65 to 75 ⁰ C for 60min or 100 to 121 ° C for 2 to 12 min; partly still using cell walliytic enzymes.

These bacterial fractions (10 ⁷ to 10 ¹² germs or starting germs / strain) are added with the addition of 2.5 parts by weight Gelatin and continuous rotation of the containers at about -40 ⁰ C frozen and at -25 ⁰ C to max. + 25 ° C lyophilized or

spray dried.

Combination vaccines: This bacterial lyophilisate and STA-R (lyophilized) are mixed so that 20 mg lectin (STA-R) is added to 10 mg bacterial lyophilisate.

(STA-R with a biological activity equivalent to 20pg of pure lectin).

The lyophilisates are mixed with physiologically indifferent excipients in a known manner to compacts, ζ. Β. Tablets, dragee cores, which include according to the invention also granules or pellets or tablets, Drageokernen, granules or pellets filled gelatin capsules are to be understood. The mixing ratio between the combination vaccines according to the invention and the tabletting excipients is between

1: 100 and 100: 1, preferably between 1:20 and 20: 1.

The granules, pellets or compacts according to the invention are used to protect against environmental influences and against the Digestive juices of the stomach in known technology provided with a coating that protects the active ingredients in the field

guaranteed from pH 1 to pH 6.

For this purpose, organic solutions of polyacrylates and polymethacrylates, vinyl polymers or Cellulose derivatives using plasticizers such as polyethylene glycol, dibutyl phthalate, triacetin, castor oil, Citric acid used for drug protection, with a intestinal juice solubility from pH 6 within 10 to 60 minutes

is guaranteed.

application

To prevent / prevent respiratory infections and activation or reactivation of the defense services in the lungs, in particular of alveolar macrophages, 12 to 40 (children / adults) tablets are administered per os in 1 to 3 months. Another form of application is also possible.

Example 2 The procedure is as in Example 1. Instead of mixing the two vaccine components (lectin and bacterial components)

in the form of lyophilisates both vaccine components are mixed in liquid form and then lyophilized together.

Claims (4)

1. A process for preparing a combination vaccine with intensified action, characterized in that from Solanum tuberosum by mechanical crushing of raw potatoes and by obtaining a pressed juice from them by low-speed centrifugation to 1 000g or by squeezing and optionally by addition of antioxidants, preferably from 0, 5 to 20 mg L-ascorbic acid per 1000 ml and subsequent centrifugation to max. 10000g for 20min in the form of the supernatant, a crude or pure preparation (crude or pure lectin), which may optionally still be pasteurized / sterilized, and lyophilized for prolonged storage and further processing / use as a vaccine component after prior deep freezing to about -40 ⁰ C. and the resulting lyophilizate, termed immune modulator "STA-R", having a biological activity equivalent to that of 2-40pg of pure lectin, with 2-20 mg of lyophilized microorganism mixture or lysate added to a mixture of by heating / heat in range from 25 ⁰ C to 13O ⁰ C for 1 to 120 minutes or at low temperature (25 ° C-35 ° C) up to 5d and partly with the addition of chemicals at very low concentrations, for example, HCHO 1: 5000, and / or by additional application of freeze-thaw effect or disintegration, for example, with U-sound at a frequency of 20 kHz and a power of about 50 watts / cm ² bzw.we Ise by very high static pressures up to about 4000atm, or by cobalt gamma rays to max. 2 ¹ SkGy and optionally inactivating chemicals in the concentration range 1: 500 to 1: 15000, or by enzymes with zellwandlytischer activity to obtain oligomeric Mucopeptide inactivated and digested current bacterial pathogens, such as from the genera Bacillus, Bordetella, Bruceila, Clostridium, Corynebacterium, Enterobacter , Escherichia, Haemophilus, Klebsieila, Listeria, Moraxella, Mycobacterium, Pasteurella, Proteus, Pseudomonas, Salmonella, Shigella, Staphylococcus and Streptococcus, which in the bacterial mixture or lysate with few strains between 1 x 10 ² to 1 x 10 ¹ * may be contained based on the starting germs or germs, and this combination of immunomodulator and bacterial mixture or lysate, which may also be added before the lyophilization with stabilizers, eventually in the form of tablets, dragees, granules, pellets or gelatin capsules in an air, water and un d acid-resistant enclosure of, for example, cellulose acetate phthalate, with a plasticizer. 2. The method according to claim 1, characterized in that is used except STA-R untreated potato juice with appropriate lectin activity. 3. Process according to claims 1 and 2, characterized in that in addition to the solid form (lyophilisate), the vaccine also liquid (for example spray, drops) can be applied. 4. The method according to claim 1, characterized in that lectins can also be used alone as immunomodulators.