US3632741A - Diethylaminoethyl dextran as an adjuvant for vaccines for active immunization - Google Patents

Abstract

DIETHYLAMINOETHYL DEXTRAN IS USED AS AN ADJUVANT FOR VACCINES FOR ACTIVE IMMUNIZATION TO PROVIDE A NEW VACCINE WHEREIN A CONSIDERABLE RAISING OF THE IMMUNITY IS ACHIEVED IN THE SUBJECTS VACCINATED.

Description

United States Patent LO 3,632,741 DIETHYLOETHYL DEXTRAN AS AN ADJUVANT FOR VACCINES FOR ACTIVE IMMUNIZATION Gunther Wittmann, Kurt Bauer, and Manfred Mussgay, Tuebingen, Germany, assignors to Farbenfabriken Bayer Aktiengesellschaft, Leverkusen, Germany No Drawing. Filed May 8, 1970, Ser. No. 35,917 Claims priority, application Germany, May 13, 1969, P 19 24 304.0 Int. Cl. (312k 5/00, 11/00 US. Cl. 424-89 8 Claims ABSTRACT OF THE DISCLOSURE Diethylaminoethyl dextran is used as an adjuvant for vaccines for active immunization to provide a new vaccine wherein a considerable raising of the immunity is achieved in the subjects vaccinated.

BACKGROUND OF THE INVENTION Field of the invention This invention relates to new vaccines containing adjuvants and more particularly to vaccines containing adjuvants comprising diethylaminoethyl dextran,

Description of the prior art The necessity of coupling antigens with adjuvants in order to achieve an improved immunity, has been well known in the prior art. There are a number of adjuvants which can be used for this purpose, among which, water-in-oil emulsions and aluminum hydroxides are most frequently used.

These adjuvants, depending on the antigen which is used with them or depending on the type of animal in which the vaccine is used, give variably favorable or less favorable results. All that the above-mentioned adjuvants have in common is that they constitute a large portion of the vaccine, namely 2550%. In addition and especially in the case of mineral oil-containing adjuvants, strong acid reactions may occur at the place of vaccination.

For these reasons, it was necessary to search for an adjuvant which:

(a) increases the immunogenic efiectiveness of antigens, that is to say the antibody formation and the immunity, and do this both in view of its beginning and its duration;

(b) is relatively slightly toxic, so that only slight reactions to vaccination occur, which lead to no permanent changes at the places of vaccination, which is particularly of importance in the case of slaughter animals;

(c) is fully effective in case of all animals;

(d) occupies little space in the vaccine volume, so that practically the entire vaccine consists of nothing but antigen. The consequence of this is, that either the dose of vaccine can be greatly reduced or that, in the case of weak antigens, considerably more antigen can be introduced with one dose of vaccine, than was possible hitherto; and

7 3,632,741 Patented Jan. 4, 1972 (e) no expensive and cumbersome manipulations are needed for production of the vaccine.

The adjuvant and resulting new vaccine of the present invention satisfy these requirements.

SUMMARY OF THE INVENTION It is accordingly one object of the present invention to provide an adjuvant and resulting new vaccine which meets the above enumerated requirements.

A further object of the present invention is to provide diethylaminoethyl dextran as an adjuvant to antigen solutions in vaccines.

Further objects and advantages of the present invention will become apparent as the description thereof proceeds.

In satisfaction of the foregoing objects and advantages the present invention provides a new vaccine in which diethylaminoethyl dextran (DEA-ED) is added directly as an adjuvant to the antigen solution.

DESCRIPTION OF PREFERRED EMBODIMENTS The present invention relates to a new vaccine in which diethylaminoethyl dextran (DEA'E-D) is added directly as an adjuvant to the antigen solution. Besides a considerable increase in the antibody formation and prolonged persistence of the antibody titers, this new vaccine also causes a considerable raising of the immunity in the case of the subjects vaccinated, as compared with application of the antigen alone.

The proportion of the diethylaminoethyl dextran adjuvant to be employed according to the present invention is about 5 to 250 mg. of DEAEDextran per ml. of innoculation serum and more preferably about 50 mg. of DEAE-Dextran per ml. of serum,

The following procedure has been found to provide good results in adding the diethylaminoethyl dextran in substance. In practicing this procedure, a proper quantity of the DEAEDextran adjuvant, within the above limits, is placed in a suitable vessel. In the case of antigens sensitive to acid, the acid pH value of the DEAED is balanced out through the addition of a proper bufier substance. It turned out to be most practical to use for this purpose a saturated tris solution. However equivalent buffering substances may also be employed. In the case of a starting pH of the antigen solution of 7.88.0, it is necessary to add approximately 2.25 ml. of saturated tris solution per 10 g. DEAED in order to obtain a pH value of 7.47.6 in the finished vaccine. If one wishes to obtain either higher or lower pH values, then correspondingly more or less tribufier is to be added. The mixture of DEAE-Dextran and the added bufiFer substance is heated in an autoclave for 2030 minutes at C. to C. for the purpose of sterilization. After complete cooling, the antigen solution is added to the solidified DEAEDextran and the mixture is kept at least 48 hours with occasional shaking at a suitable temperature. In this time, the DEAE-Dextran usually is dissolved. After this is the case, the vaccines are shaken for 1 hour and they are then allowed to stand until their use, once more for at least 24 hours at a suitable temperature.

One can use as antigens all substances which are immunogenically effective in a purified, partly purified or TABLE 2 The formation of neutralizing antibodies in the case of guinea pigs after inoculation with mouth and hoof disease vaccines with varying quantities of DEAE-D DEAE-D ND values of the scrums per ml. vaccine 4 weeks after vaccination 5 mg. 1:151:113- 1.63 15 mg. 1:641:2801:151 30 mg. 1:195-1:1,0801:451 45 mg. 1:235-1:1,5001:937 60 mg. 1:300-1:1,5001:674 Control Negative-lz3l-1z10 1 Extreme values. 3 Mean value from the individual titers of 60 guinea pigs. 3 Inactivated virus without adjuvant.

6 aminoethyl dextran adjuvant is present in an amount of about 5 to 250 mg. per ml. of innoculation serum.

4. Vaccines according to claim 3 wherein said diethylaminoethyl glextran adjuvant is present in an amount of about mg. per ml. of innoculation serum.

5. Vaccines according to claim 3 wherein said antigens are sensitive to acid and the pH value of the diethylaminoethyl dextran is balanced out by the addition of a buffer substance.

6. Vaccines according to claim 5 wherein sufficient butter is added to obtain a final pH of 7.4-7.6 in the finished vaccine.

7. Vaccines according to claim 6 wherein the buifer substance is saturated tris solution.

8. Vaccine-s according to claim 7 wherein the bufier solution and diethylaminoethyl dextran are heated at C. to C. prior to addition to the antigen.

References Cited UNITED STATES PATENTS 2,908,614 10/1959 Muggleton et a1. 42488 X SHEP K. ROSE, Primary Examiner US. Cl. X.R. 42488, 180, 361