DD291007A5 - PROCESS FOR PREPARING VACCINES WITH INCREASED EFFICACY - Google Patents

Abstract

The invention relates to a process for the preparation of vaccines with increased efficacy and relates to the field of pharmacy. The object of the invention to achieve a greater immunization effect with vaccines with the aid of new adjuvants was achieved by carrying inactivated virus or bacterial material with polysaccharide derivatives carrying functional groups of the general formula wherein R 1 CH 2, C 2 H 4, R 2 H, CH 3, C 2 H 5 and R 3 H, CH3, C2H5, are converted to pharmaceutical preparations. Areas of application of the resulting vaccines are human and veterinary medicine as well as agriculture (animal production). {Inactivated vaccines; adjuvants; polysaccharide derivatives; Newcastle Disease Virus; Virus vaccines; Immunity; physiological compatibility; pharmaceutical-technical effort; applicability; Human and veterinary medicine}

Description

Field of application of the invention

The invention relates to a process for the production of vaccines with increased efficacy, which can be applied in the field of human or veterinary medicine, pharmacy and in agriculture (animal production).

Characteristics of the known state of the art

It is known that many viral or bacterial antigens when injected into an organism do not elicit a measurable or only a slight immune response, so that they can not readily be used as a vaccine. However, the antigenicity of viral or bacterial materials can be enhanced by the addition of adjuvants. Numerous substances have been proposed in the literature which can be used as adjuvants. The most commonly used so far are aluminum hydroxide, mineral and vegetable oils and metabolites of bacteria (THEIN, Vet. Med. Nachr. 59 [1988]). Very good adjuvant effects are achieved by emulsifying aqueous antigen solutions with mineral or vegetable oils. However, Solcho emulsions, especially mineral oil emulsions, are poorly tolerated locally. After vaccination of oil emulsion vaccines, macroscopic changes at the injection sites can be detected up to 28.1 weeks post vaccinationem, which may lead to regulations for meat-producing reasons in livestock. Furthermore, oil emulsion vaccines are relatively viscous, which makes handling them difficult. In addition, suitable for the oils suitable emulsifiers.

Unsatisfactory with most known adjuvants is generally their often poor efficacy or inadequate physiological compatibility.

Object of the invention

The object of the invention is therefore to make available to the pharmaceutical industry for human or veterinary purposes vaccines. 'It was characterized by increased efficacy and good physiological tolerability. At the same time, cit>. pharmaceutical-technical effort to minimize their production and to ensure good applicability.

Explanation of the essence of the invention

It is an object of the present invention to provide a method which results in improved efficacy vaccines with the aid of new adjuvants, i. For the same amount of antigen better immunization than without adjuvant or with conventional adjuvants.

According to the invention, the object is achieved by mixing inactivated virus or bacterial materials with sterilized solutions of polysaccharide derivatives which are functional groups of the general formula

-0-R - N '

where R ₁ = CH ₂ , C ₂ H ₄ , R _} = H, CH ₃ , C ₁ H ₆ and R ₃ = H, CH ₃ , C ₂ H ₆ . For example, products based on cellulose, hemicellulose, agarose and starch may be used as polysaccharide derivatives, the content of functional groups per polysaccharide basic unit being 0.05 to 1.0. Diethylaminoethylcellulose proved to be especially suitable. The degree of polymerization of the polysaccharide derivatives according to the invention can be varied within wide limits, but should not exceed 500, so that the viscosity of the resulting vaccine does not become too large. For the production of the vaccine it is advantageous to proceed from a 1 to 10% solution of the polysaccharide derivative in physiological sodium chloride solution, the dio is first sterilized, for example by autoclaving at 121 ^° C. Subsequently, the inactivated virus or bacterial material is mixed with the adjuvant solution in such a ratio that the adjuvant concentration in the vaccine is 0.5 to 5%. The vaccine is administered in doses of 0.2 to 2 ml, each containing 2 to 20 mg of adjuvant. The vaccines prepared according to the invention are distinguished by very good general and local compatibility, good applicability, high antibody formation and thus also high protective action. The proposed adjuvants are characterized by an enhancing effect of the antigen, innocuousness for vaccination and ease of preparation or use.

Although it is known that various polysaccharides have immunostimulatory properties, which are mainly attributed to dio β 1,3 and β-1,6 linkages of glucose, and must be given certain conditions in terms of molecular weight and solubility, but was due to this Knowledge can not be expected that the invention proposed as adjuvants polysaccharide derivatives have a special immunostimulating effect. Rather, it has been found that comparatively tested solutions of hydroxyethyl cellulose and Natriumcarboxymethylcnllulose show no positive effect

 The invention will be explained in more detail with reference to the following examples.

Ausführur.gsbelsplel Preparation of an Inactivated Vaccine Against Newcastle Disease (ND) of Chickens For the preparation of vaccine samples, raw virus material of the strain "La Sota" inactivated with 0.1% formalin,

used. The virus titer before inactivation was 10 ⁹ ' ¹ EID _M / 0.1 ml. Per vaccine dose was a virus amount of 80μΙ

used.

Adjuvanszubereitung: It wui Jo made a 5% solution of diethylaminoethylcellulose, adding 5g of dry matter

boiled with 100 ml of physiological saline and then autoclaved at 121 ⁰ C for 30 minutes.

Dia diethylaminoethyl cellulose was used by homogeneous reaction of regenerated cellulose with

2-Chlorethyldiethylamin hydrochloride prepared in 10% sodium hydroxide solution and then isolated by precipitation with E'hanol.

The product has a degree of substitution of diethylaminoethyl groups of 0.4 and is water-soluble. The Degree of polymerization is about 220. The vaccine preparation was done by mixing 120ml of inactivated virus material with 80ml of adjuvant solution. Dio Adjuvant concentration is thus 2% in the vaccine, d. h., in 0.5 ml vaccine (1 vaccine dose) are 10mg of the adjuvant included. Efficacy evaluation: The'. Incubation of chicks was done at 31 days of age with one dose per animal. 3 weeks later became one Test infection performed with a velogenic ND strain. The experiment was completed 14 days after infection. The Efficacy testing was carried out according to the criterion of protection rate (healthy surviving animals) and the serological response

(Hemaggiutinationshemmungsreaktion) evaluated.

Several vaccines were used for efficacy testing.

Protective Raton after test infection vaccine

1. ND vaccine with DEAE-cellulose as adjuvant

2. ND vaccine without addition of adjuvant

3. ND adsorbate vaccine "Dessau" ·

4. Controls (no immunization)

protected/ protection rate confidence total % range ρ = 0.05 26/26 100 86.77-100 18/26 69.23 48.21 to 85.67 17/26 65.4 44.33 to 82.79 0/15 0 0 to 21.80

Result of the serological examination

vaccine

xTKZbefore immunization

xTKZ 20Tago after immune.

χ TKZ

14 days after

infection

1. ND vaccine with DEAE-Collulosa as adjuvant

2. ND vaccine without adjuvant

3. ND adsorbate vaccine "Dessau" ·

4. Controls (no immunization)

TKZ ₌ Titerkennzahl without titer 0 1: 1 1 1: 2 2 1: 4 3 5? TKZ Arithmetic mean of the titer identifier n.u. not examined

7.26 2.88 3.62 0

7,33 10,75

8.5 n.u.

Claims (6)

1. A process for the preparation of vaccines with increased efficacy by adjuvant, characterized in that inactivated virus or bacterial materials are reacted with polysaccharide derivatives, the functional groups of the general formula -QR ₁ -N ^^^ where R, = CH ₂ , C ₂ H ₄ , R ₂ = H, CH ₃ , C ₂ H ₅ and R ₃ = H, CH ₃ , C ₂ H ₅ . 2. The method according to claim 1, characterized in that are used as Polysaccharidderivate products based on cellulose, hemicellulose, agarose or starch. 3. Process according to claims 1 and 2, characterized in that the degree of substitution of the polysaccharide derivatives is 0.05 to 1.0 and the degree of polymerization is 20 to 500. 4. Process according to claims 1 to 3, characterized in that is used as the polysaccharide diethylaminoethylcellulose. 5. The method according to claim 1, characterized in that as inactivated viral antigen raw virus material of the strain "La Sota" is used and the virus titer before inactivation ä10 ⁷ EID / 0.1 ml. 6. Process according to claims 1-4, characterized in that the polysaccharide derivative is sterilized as a 1-10% solution in physiological saline, preferably by autoclaving at 121 ⁰ C, and then with the inactivated virus or bacterial material in the volume ratio 1: 10 to 10.1 is mixed.