DE2425917A1 - VETERINARY MEDICINAL PRODUCTS AND METHOD FOR MANUFACTURING IT - Google Patents

Description

BEECHAM GROUP LIMITED, Brentford, Middlesex and THE MINISTER OF AGRICULTURE FISHERIES AND FOOD, London / England

"Veterinary medicinal products and process for their manufacture" Priority: June 1, 1973 - Great Britain - Number 26 251/73

The invention relates to a veterinary medicinal product for prevention and control of Newcastle disease in poultry (atypical avian influenza).

Newcastle disease is a viral infection that, because of its extremely high risk of infection, its numerous forms, in which it appears and is notorious for its widespread use. The most essential method of protection against this disease consists in the use of inactivated Newcastle disease vaccines, which are administered together with an excipient (See Lancaster (1966) "Newcastle Disease a review 1926-1964"). This publication mainly deals with the inactivating Agent, its concentration and the effect that numerous formulations have in terms of duration of immunity. About the choice Little has been published of the excipients. A typically used auxiliary is the "Alhydrogel". 1972 is about the

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Function of an oily adjuvant in inactivated vaccines to protect against Newcastle disease has been reported (cf. W.H. Allan "in Agriculture" 79: 413-420 (1972)).

Although such treatments are widely used, they are seldom fully effective because the known formulations do not sufficiently stimulate the production of antibodies in poultry to produce complete immunization. This problem of potentiating an antibody response has occurred in other areas, and Woodhour et al., In the journal Proc. Soc. Expl. Biology and Medicine, 131 (1969) p. 809, reports that the addition of a synthetic double-stranded polyribonucleotide to a flu vaccine in the so-called "Adjuvans 65" increased the antibody response in guinea pigs and monkeys.

It has now been found that the antibody response in poultry in the case of inactivated Newcastle disease viruses, significantly due to the simultaneous administration of double-stranded ribonucleic acids of natural origin is reproduced.

The invention thus relates to veterinary medicinal products which contain inactivated Newcastle disease viruses and auxiliaries and either

(a) a double-stranded ribonucleic acid of natural origin or its salt or

(b) a double-stranded derivative of a double-stranded ribonucleic acid of natural origin or their salt are labeled.

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The term used in the description and claims "Double stranded" refers to the property that makes two Ribonucleic acid molecules by means of a hydrogen bond between complementary bases are linked on each molecule, regardless of whether other forms of bond exist between the two Molecules are present.

The term "double-stranded ribonuclear acid of natural origin" means any double-stranded ribonucleic acid derived from Naturally occurring sources can be isolated, and excludes biosyncnetic double-stranded ribonucleic acids (hereinafter abbreviated to RNA) from, such as polyinosinic acid; Polycytidylic acid (Poly IJ Poly C), polyadenylic acid: polyuridylic acid (Poly Ai, Poly U) and Polyguanidylic acid: polycytidylic acid (Poly Gi Poly C). Under the specific sources of a double-stranded ribonucleic acid of which have been reported to be the virus particles that are present in certain infected fungi are present, e.g. from certain Penicillium species, namely Penicillium chrysogenum, Penicillium stoloniferum, Penicillium funiculosum and penicillium cyaneofulvum, as well as Aspergillus niger and Aspergillus foetidus. In addition, such ribonucleic acids are found in the yellow virus of the sugar beet, in the rice-dwarf virus, in reovirus 3, in cytoplasmic insect polyhedrosis virus, in the replicative phage form of MS2 from Escherichia cöli, in the replicative phage form of MU9 from Escherichia coli and in the replicative phage form of Pseudomonas aeruginosa.

The term "double-stranded derivative of a double-stranded ribonucleic acid of natural origin "means any double-stranded ribonucleic acid of natural origin that is chemically

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see or subject to biochemical, e.g. enzymatic, conversion which change the primary and / or secondary and / or tertiary structure, provided that the derivative obtained retains a substantial degree of base pairing ability between complementary strands. Examples of such derivatives are the N-oxides described in GB-PS 1 284 156, which are in the German Offenlegungsschrift 2 201 51J described alkali-modified, double-stranded ribonucleic acids or those described in German Offenlegungsschrift 2 256 78I with formaldehyde modified double-stranded ribonucleic acids.

Examples of suitable salts of double-stranded ribonucleic acids or double-stranded derivatives are alkali metal salts such as Sodium or potassium salt, the ammonium salt and substituted amine salts, and complex cations disclosed in German Offenlegungsschrift 2 253 977 and British patent application 55 425/71 are described.

The term "adjuvant" used in the description and in the claims includes all substances which, when taken together with formulated into a vaccine are known to increase the antibody response in vaccines. Examples of such auxiliaries are naturally occurring or mineral oils that, when formulated with the vaccines, provide water-in-oil emulsions. A An example is the adjuvant known under the name "Adjuvans 65", which is a water-in-peanut oil emulsion. Further Examples of excipients are absorbents, such as potash alum, which has an absorbing and precipitating effect on proteins possesses, mineral absorbents such as aluminum, calcium

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or magnesium hydroxides, which can be formulated to contain the viruses in an insoluble gel. For example is the adjuvant "Alhydrogel" an aluminum hydroxide gel,

Any strains of the Newcastle disease virus are suitable according to the invention. Inactivated Newcastle disease viruses are commercially available or can be isolated from known sources and inactivated, for example, with formaldehyde. A mild virus strain is preferably used. A particularly suitable strain is the Ulster strain, which was first isolated by McPerran and co-workers (cf. "Vet. Rec. ^1t 82 (1968), pp. 589-592).

When administering the veterinary medicinal product according to the invention, can the double-stranded ribonuclear acid (ds-RNA) in the excipient as a separate preparation from the composition of Virus plus Adjuvant can be administered, but preferably a single preparation of the viruses, the adjuvant and the ds-RNA is administered. The agent is usually injected into the poultry. If the two preparations are administered separately, they will be better Results obtained when the oil-in-water emulsion injected a solution of the ds-RNA at the same site as the oil-based vaccine will. If two injections are given in different places, there will be less effect.

If a single preparation is used, it can be done either by emulsifying the two ingredients individually and then Mixing with one another or by adding the ds-RNA directly to the aqueous virus antigen solution and then emulsifying be generated. It has been found that the last-mentioned method

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usually have higher levels of antibodies after vaccination supplies.

The weight of the introduced ds-RNA per dose varies with the type of virus and the adjuvant used, but a suitable level is in the range from 0.1 to 1.0 mg ds-RNA per dose.

The following examples illustrate the invention. The following materials are used in these examples and also applied the general process steps indicated.

poultry

There are always sensitive to Newcastle disease, 3 to 9 week old chicks used. These chicks come from a flock that is free from Newcastle disease, the chicks have not been vaccinated and have also been examined prior to vaccination to ensure that all animals are negative against the Newcastle disease vaccine (abbreviated NKI) are.

Vaccine Two types of vaccine are used:

(a) To produce the viruses, the Ulster strain of the Newcastle disease vaccine is used, which was first isolated by McPerran and co-workers (cf. "Vet. Rec." 82 (1968), pp. 589-592). This preparation is designated "2C". Sixty 9-day-old eggs already containing embryos are inoculated via the allantoic cavity with 0.2 ml of a 10 ^-1 virus suspension from a redissolved freeze-dried ampoule. After 4 days of incubation at 37 ⁰ C,

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the noble eggs *, and the allantoic praise

(AAF)
water / taken. The virus suspension is cleared by centrifuging for 10 minutes at 2000 revolutions per minute on an "MSE-Super-Multex". The supernatant is maintained at a temperature of -70 ⁰ C before the treatment. The viruses are inactivated by adding formaldehyde to achieve a final concentration of 0.04 percent. The mixture is incubated for 18 hours at 37 ° C. and poured into a fresh bottle to ensure that the viruses and the formaldehyde are completely mixed. A 5-fold passage inoculation of embryos is carried out with the fluids in order to ensure complete virus inactivation.

(b) A commercially available inactivated Newcastle disease vaccine in an oily excipient is used immediately in several examinations. Manufacturer of this vaccine is the Istituto Zooprofilattico Sperimentaüe della Lomberdia e dell Emillia, Brescia, Italy. This vaccine hereinafter referred to as the commercial vaccine.

Double stranded ribonucleic acid

A highly purified fungus obtained from a virus particle ds-RNA, a modified polyquaternary complex of this ds-RNA, and a phage form of this ds-RNA are all created as aqueous preparations from freeze-dried material. All concentrations are measured spectrophotometrically by absorbance determined at a wavelength of 259 mu.

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Excipients

Two types of excipients are used in the examples: "Adjuvant 65", which consists of 86 percent peanut oil, 4 percent aluminum stearate as a stabilizer and 10 percent of a sorbitan ester of a higher molecular weight fatty acid as an emulsifier ("Arlacel A"), and a less viscous preparation which has been described by Thompson and coworkers in the journal ^M Vet. Rec. "85 (1969), pp. 8I-85. This incomplete Freund-type excipient consists of 90 percent mineral oil (" Dracneol No. 6v " ) and 10 percent of the aforementioned sorbitan ester of a higher molecular weight fatty acid ("Arlacel Α ^Μ ). The ratio of vaccine to excipient is specified in each experiment.

serology

The serum NKI-antibody levels are determined by the haemagglutination inhibition (HI) test, where a "Canalco Car Titratör ^III-n (Fisons Scientific Apparatus, Loughborough, England) is used. Volumes of 0.05 ml of double dilute Sera are tested against 4 HA 50 units of the formaldehyde-reacted NKI antigen. One percent of chick erythrocytes washed 3 times are added to an equal volume. After 1 hour at 4 ° C., the results are read by the respective operator. The individual titers. are expressed as the reciprocal of the logarithm of base 2 of the 50 percent serum dilution endpoint, and the results of each group are expressed collectively as the "mean titer-log".

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Immunity test

The chicks are infected on the 42nd day after vaccination by spraying with the Essex ^r 70 strain of the NKI. A hand-operated aerosol generator is used to generate droplets with an average dry particle diameter of 3 H.

1 ml of the AAF containing the virus is distilled with 50 ml

Dilute 8 ½ of water so that a final concentration of 10 '^ / ml EID, - ₀ is obtained. The observations are carried out twice a day for 14 days. Chicks that are dying are killed and included in the total mortality rates. (ETD ₁ -, means the concentration of virus that is able to infect 50 percent of the fertilized eggs when injected).

Example 1

From the virus particles found in Penicillium chrysogenum ATCC 10002 a double-stranded ribonucleic acid (ds-RNA) is obtained.

90 9 week old chickens are divided into 4 groups of 20 animals each and 1 group of 10 control animals. The 2C vaccine is with the "Adjuvans 65" of the .ds-RNS in an amount of 0.62 mg per dose and with saline to 4 different preparations mixed. The individual formulas are given in Table I

indications given. The preparations are in a final Inoculated intramuscularly at a dose of 0.5 ml per chicken. All Results are shown in Table I.

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Table I.

Mean anti-haemagglutination antibody levels after vaccination (mean titer-log)

Group treatment ratio of 14 days 21 days 55 days

Vaccine + ds-RNA for excipient immunity test Mortality in %

• το co oo

aqueous 2C vaccine 1.90 2.50

2.60

2?

aqueous 2C vaccine + 0.62 mg ds-RNA 1.77 2.70

2.72

2C vaccine in "AdJuvans 65 "2.31 2.85 3.65

20th

2C vaccine in "Adjuvant 65" + 0.62 mg ds-RNA in "Adjuvant 65" 4.20 6.18 6.55

Con- not vaccinated trolls

<1.00 <1.00 <2.00

100

The addition of aqueous ds-RNA in an amount of 0.62 mg per dose has very little, if any, effect on the haemagglutination inhibition antibody response to the 2C "Newcastle disease vaccine in poultry. The same amount of ds-RNA, however, shows one marked stimulus in antibody level when given with 2C vaccine in "Adjuvant 65" and is remarkably "higher than other groups (p ^ 0.001) at Ik, 21 and 35 days after vaccination.

These differences in antibody levels are consistent with the percent mortality from infection of the chicks with the NKI on day 42. The only group that has been fully protected is those that received the 2C vaccine and 0.62 rag ds RNA in "Adjuvant 65". It can be seen that the inclusion of ds-RNA in an oily mixture of excipient and vaccine has increased the mean logarithm of the titer by more than k units, ie by 16 times the amount of the antibodies, and that the antibodies appear earlier and last longer stay when the ds-RNA is incorporated into the vaccine.

Example2

ΙΛΟ 9 week old chickens are divided into 7 groups. the Groups 1 through 4 will receive 2G vaccine cancellations and the groups 5 to 6 each formulations with the commercially available vaccine, with Freund's Incomplete Adjuvant. The 7th group is the non-vaccinated control group. In this attempt it will be as in example 1 obtained ds-RNA administered in an amount of 0.31 mg per dose. The comparable formulations and results are out

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Refer to Table II. Each chicken has 0.5 ml intramuscularly of the respective solution.

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Tabel

II

- 13 -

Mean anti-haemagglutination antibody levels after vaccination (mean titer log)

Group treatment ratio of 14 21 28 35 ^ 2

Vaccine + ds-RNS days days days days days • ■ to the excipient

Immunity test mortality in %

· Aqueous 2C vaccine

aqueous 2C vaccine + 0.31 mg ds-RNA

2C vaccine in $ 50: $ 50 "Adjuvant 65" + PBS in "Adjuvant 65"

2C vaccine in "Adjuvant 65" + 0.31 mg ds-RNA in "Adjuvant 65"

commercial vaccine (oil emulsion + PBS in one, one oil adjuvant

commercial vaccine + 0.077 mg ds-RNA in an oil adjuvant 2.05 2.00 2.15 1.95 2.05

2.00 2.00. 2.00 2.00 2.00

2.05 2.40 '3.00 2.55 2.80

3.68 4.20 5.21 5.05 5.45

2.60 3.50 3.25, 3.75 4.84

3.00 4.4o 5.55 5.55 6.26

50 70

30 0

not vaccinated control

2.00 2.0 2.0

2.0

2.0

100

(PBS = phosphate-buffered NaCl solution)

In this experiment, smaller amounts of ds-RNA are grounded with the NKI in "Adjuvant 65" and also in the oily one already described Adjuvant studied. Although emulsification of the 2C vaccine in the "Adjuvant 65" compared to an increase in the HI antibody level that produced with an aqueous 2C vaccine, a much greater increase is achieved when 0.51 mg of ds-RNA is present alone be administered to these formulations. The immunity test shows a similar picture to the previous test, with only one group receiving the 2C vaccine and the ds-RNA in "adjuvant" 65 "has been treated, full protection is obtained.

A similar increase in the antibody level and protection against infection with NKI is found when 0.077 mg ds-RNA is in a thin oily excipient can be combined with a commercially available vaccine. During the experiment, the equal amounts of the excipient with the sole or combined Vaccine treatments are given to ensure that the observed reactions not only on one alone Nigen increase in the amount of excipients are based.

Example 3

100 6 week old chickens' are used in this experiment, and divided into 5 groups.

Group 1 receives a commercial vaccine in an oily one Excipient, the groups 2 to 4 the 2C vaccine in the incomplete Freund's adjuvant, and group 5 remains unvaccinated.

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- ¹ ^ - 2 i 25917

trolls.

The ds-RNA obtained as in Example 1 is used in an amount of 0.077 mg immediately with one of the 2C vaccine preparations (for group 3) mixed, while for group 4 the ds-RNA im Aid is administered as a separate injection. The comparative formulations and results are shown in Table III. The preparations are as in the two other examples inoculated.

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Table III

Mean anti-haemagglutination antibody levels after vaccination ■ (mean titer log)

Group treatment ratio of 14 21 28 35. 42 49 56

Vaccine + ds-RNA days days days days days days days to the excipient

4b

commercial vaccine

2C vaccine. 20 ^: $ 80 in oil with PBS in an oil adjuvant

2C vaccine $ 20: ¹ with O, 077 mg of ds-RNA in oil

2C vaccine 20 ^: in oil 0.077 mg ds-RNA in oil

4.6 5.5 4.8 5.7 ■ 4.5 5.2 5.5 2.1 2.3 2.7 3.2 3,

4.2 4.2

4.8 6.1 7.8 7.7 7.6 8 ', 4 8.1 2.5 3.2 3.8 3.8 3.6 4.6 4.2

control

not vaccinated

2.0 2.0 2.0 2.0 2.0

2.0 2.0

a = vaccine and ds-RNA given in a single injection; b = vaccine and ds-RNA given in separate injections.

Adding ds-RNA to an oily emulsion delivers the vaccines a remarkable increase in HI levels, which lasts longer than vaccines based on antibodies alone and emulsion based. As can be seen from Table III, are the HI antibody levels with the combined emulsions were significant higher than in the case of the injections, in which the oily emulsions of the 2C vaccine and that of the ds-RNA are injected separately. the by an equivalent "volume of the commercially available vaccine antibody titers generated, although greater than those obtained by separate injections of the 2C vaccine in oil on the one hand and the ds-RNA in oil on the other hand have been obtained, yet lower than that of the combined emulsion.

Example 14

Different dose levels

The aim of this experiment is to investigate how little ds-RNA is required is to determine the reaction of the excipient. the double the amounts of ds-RNA obtained as in Example 1 of 0.01 to 0.8, mg are in the thin oily described earlier Emulsion made (4 volumes of incomplete Freund's adjuvant to 1 part by volume of aqueous phase) at a dose of 0.5 ml per chicken. Each of these preparations is then administered intramuscularly alongside an equal volume of a commercially available oil-based vaccine injected into the right pectoral muscle. The chickens vaccinated as controls receive emulsified saline and vaccine in the same volume. For determining the serum HI antibody level will blood the chickens for 18 weeks

QO tapped. Then they are aerosolized with 10- "EID's

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Northampton strain of the NKI injected. The results sirrcr off Table IV can be seen.

Table IV

log,
C. 3 of the mean antibody titeri 21 28
after 35
to the .42 49
Vaccinate 6.2 56 treatment 7th 14th
Days Λ9 5.6 6 _r 0 5.2 Vaccine +
0.08mg 6s-
RNA 1.0 4.0 6 _f 2 _4/7 4.7 5.0 5.1 Vaccine +
0.04mg ds-
RNA 1.0 5.8 4.3 5.8 4.7 6.1- *, 9 Vaccine +
0.02mg ds-
rns 1.0 5.1 5.7 4 _; 4th 5.8 5.8 4.5 Vaccine +
0.01mg ds-
RNA 1.0 5.6 *, 9 5.5 4.6 5.1 <2.0 4.2 vaccine 1.0 5.2 <2.0 <2.0 <2 _; 0 not
vaccinated <2.0 <2 _f 0

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Table IV continued

log ₂ des mean antibody titer 98
to the 112
Vaccinate Vii ^ us Infection
mortality
in % treatment 70
Days 84
after ■ 5.6 5; 4 0 Vaccine +
0.08 mg ds-
RNA 6.0 5.5 5/2 4.3 14.2 Vaccine +
0.04 mg ds-
RNA 5.9 5/1 5.1 4.8 0 Vaccine +
0 * 02 mg ds-
RNA .6.5 6.1 5.0 4.8 0 Vaccine +
0.01 mg ds-
RNA 6 _; 8th 7.4 4.3 3.6 11.1 vaccine V 4.3 <2 _f 0 <2.0 100 not
vaccinated <2.0 <2.0

Example 5

Using the identical formulations and the same procedures as in Example k , two other ds-RNA preparations are examined for comparison purposes with regard to their adjuvant potential with commercially available NKI in oil. One of these ds-RNAs is a polyquaternary complex of the RNAs used in Example 1 and can be prepared according to Example 2 of German Offenlegungsschrift 2,253,977.

The second ds-RNA is a purified ds-RNA, originally from the replicative intermediate of the sus-3 mutant of the f ₂ coliphage

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2 A? 5 91 7

been isolated i, st. The HI antibody levels and the results of the aerosol injection rait NKI are shown in Table V.

νο 0 VO U Ό ft in φ • φ CO κ \ bO O ιη Φ W. ECO C. O Ό • ^ α ν. ft OS • Η pd ^ t CVl H χα ft «K ιη " Φ E. ι M. V Ο \
■ a · -P CQ t- M. O CD κ O ■ σ φ CVJ E. V W. ιη " CTrH ιη ~ CVJ C. φ CVl E. VO φ O O • Η φ
■ a Η " ftO ω CvT -P CVl O ιη E. co V Φ «Ν H ft ιη ιη ιη Pooh ιη ο • ν is! ο
Cj CVl • Η CO ο α ιη VO CVl 00 O C.
φ ιη CvT U φ , _, CO V φ bO ν -P
.ρ CVI CD ο "ε CVl
V. CQ ιη ι-ί Φ CV? O TJ γΗ CVI
U) ί- V O ο O ιΗ H H O ^. Ο cvT ι-ί V -P W. + «Μ "** ft W. + ι E. Yy • Η ο ε O Φ C. +3 Ci I faO CD ω ιη O C - «^ CVl + 3 +> φ Ul I CQ E V <r-I O rH HO ftO • Η E ^ α H ^

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- 21 Continuation of Table V

treatment logp of the middle
70 84
Day· . after 5.9 Antibody titer
98 112
vaccination 4.8 Virus infection
tion
Mortal
speed in % Vaccine +
0.125 mg
polyquaternary
Complex of "
ds-RNS 5/6 6.0 V 3.7 14.2 Vaccine +
0. 0Ö2 mg one
Phage form
the ds-RNA 4.9 5/0 5.6 <3.0 11.1 Vaccine "+
NaCl in oil
solution 3.8 5; 0 0 not vaccinated <2.0 . <3.0 100

Example 6

The effect of combined preparations of ds-RNA and vaccine as the only solution for injection is examined in two ways:

(a) Emulsifying the two ingredients, respectively, and then mixing of the two emulsions and

(b) adding the ds-RNA to the aqueous virus antibody immediately and subsequent emulsification.

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(a) The concentrations of the RNA obtained as in Example 1 in saline are ^{emulsified in a ratio of 1: 1 or 1: 4 with the oily incomplete Freunds 1} adjuvant, so that an amount of 0.125 mg in 0.25 ml is finally obtained . These preparations are then either administered as separate injections with 0.25 nil of an oily emulsion from commercially available NKI or mixed in the same volume and administered intramuscularly as a single injection in an amount of 0.5 ml per chicken. The serum HI antibody level up to 10 weeks after vaccination and the percentage mortality after aerosol infection with NKI are shown in Table VIa.

(b) To maintain a constant antigen level of the virus in the vaccines to maintain, with only a single emulsification stage taking place, the allantoic virus preparation is 2 hours at Centrifuged 50,000 g and then resuspended in a saline solution in 25 percent of the original volume. the 2C strains of Neweastle disease virus are alone and with the Addition of ds-RNA for a dose of 0.08 mg per chicken emulsified. Separate emulsions are also produced in each case and on opposite ones Places injected. The commercially available Newcastle disease vaccine is also included for comparison. 10 animals each 0.5 nil of all preparations are injected intramuscularly. The results are from the table VIb can be seen.

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Table Via

treatment Oil volu
men, % 50 Meet
chung +) log ₂ of the mi dead Antibody titer 42
to the 49 56
Vaccinate ul 70 Virus-In
fection
(Mortal
speed in %) commercial practice
cher vaccine
material , 50 - 14th 21
Days 30th
after 35 3.3 2.5 3.8 • ul 20.0 commercial practice
cher vaccine
fabric + ds-RN S. .50 separated <3.0 4.1 3.4 ul 3.2 5> 7 3.5 3.7 11.1 commercial practice
cher vaccine
+ ds-RNA f 80 together ^ 3.0 4.7 4.6 ul 3.8 4.4 4.7 4.1 16.6 commercial practice
cher vaccine
+ ds-RNA f 8o separated <3.0 5.0 4.9 3.5 5.0, 4.8 5.0 4.5 0 commercial practice
cher vaccine
+ ds-RNA f - together <3.0 5.7 5.2 4.5 4.8 4/6 4.2.0 4.8 33.3 not vaccinated - 43.0 6.2 5.7 5.2 <2.0 <2.0 <2.0 100 <3.0 <3.0 <2.0 42.0

ul = illegible

rf) = combined doses delivered as a single intramuscular injection of 0.5 ml administered in an equal volume after mixing; separate doses presented as two intramuscular injections of 0.25 ml each administered at opposite locations.

Table VIb

log ₂ des mean antibody titer 2.7 35
to the 2 42
Imofea 49 56 2 treatment 14th
Days 21 28
after 7.8 3, 7th 3.3 4.2 1 2C vaccine
in oil 2.1 2/5 3.8 7, 8th 7.6 8.4 8th, 2 2C vaccine
with. 0.08mg
ds-RNA in oil
(together) 4.8 6.1 4.8 7th 4.6 K 5 2C vaccine
in oil;
0.08mg ds-
RNA in oil
(separated) 2.5 3.2 2.0 5, 0 5.2 5; 0 commercial practice
cher NKI 4 _; 6th 5.5 2, 2.0 2.0 2, not vaccinated 2.0 2.0

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Claims (1)

- 25 patent claims Z 4 Z for I / 1. Veterinary medicinal products, marked by a Content of inactivated Nevjcastle disease viruses, excipients and either (a) a double-stranded ribonucleic acid of natural origin or its salt or (b) a double-stranded derivative of a double-stranded ribonucleic acid of natural origin or their salt. P. veterinary medicinal product according to claim 1, characterized by a content of such a double-stranded ribonucleic acid which is derived from virus particles which are found in infected strains of Penicillium chrysogenum, Penicillium stoloniferum, Penicillium funiculosum, Penicillium cyaneofulvum, Aspergillus niger or Aspergillus the replicative phage forms of MS2 from Escherichia coli, from MU9 from Escherichia coli or from Pseudomonas aeruginosa. 3. Veterinary medicinal product according to claims 1 or 2, characterized through a content of naturally occurring or mineral oils as auxiliary materials. 4. Process for the preparation of the veterinary medicinal product according to Claims 1 to 3, characterized in that one is inactivated Newcastle disease virus emulsified in an excipient can be with either (a) a double-stranded ribonucleic acid of natural origin ^or 409851 / 1114- (b) a double-stranded derivative of a double-stranded ribonucleic acid of natural origin, which may be emulsified in an excipient, mixed and, if no emulsification can be carried out, then the mixture is emulsified in an excipient. 5. The method according to claim 4, characterized in that the double-stranded glbonucleic acid or its derivative is added directly to the aqueous virus and then emulsification with the virus is carried out. 409851/1 1 U