DE1617627A1 - Process for the preparation of a storage-stable, combined vaccine against measles and smallpox - Google Patents

Description

Process for the preparation of a storage-stable, combined vaccine against measles and Poeken

This invention relates to a live combined measles virus and smallpox vaccine, and more particularly to one Preparation that has a considerable shelf life,

Hasern (Bubeola) and smallpox both mean for themselves an extremely important health problem to most of the world's peoples. Highly effective vaccines have been developed against each of these diseases, but they are usually administered separately. Significantly easier and cheaper it would be if the two vaccines could be given together in a single dose. this would particularly apply to the underdeveloped countries of the world, where the medical facilities

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are restricted for the administration of vaccines.

Previous research indicated that measles and pooken vaccines could be obtained using a syringe as a means for the Injections in humans can be administered together. This way of working, however, requires that the separate, liquid measles and smallpox vaccines be immediately available be mixed upon administration. It is obvious that the handling of the vaccines at the time of use considerably relieved and in particular the risk of contamination and error in handling on one The minimum level could be reduced if the. both Substances could be mixed beforehand and stabilized in mixed form by drying

The present invention relates to the teohnology of the premisohene of measles and pooken vaccines, distribution into individual containers and stabilizing the activities of both components through the lyophilization process. More specifically, it has been found that it is possible to use both the measles and pooken components of the vaccine in a mixed form without substantial loss to dry immunizing activity · Such misohten Vaccines are intended for injection in humans under * Using a syringe with either a sloping or a vertical head, also the intradermal or subcutaneous injection

$ 103,813 / $ 159

tion with the hypodermic injection is acceptable. The sloping one Syringe head allowed, 0 »! ml of the combined vaccine mixture To notify you internally. The vertical head Allowed if he is using a cuff that is appropriate for the The distance between the nozzle and the skin ensures that 0.1 ml of the combined vaccine is distributed in the intradermal area and from 0.4 ml to the deeper Tissue. When used without a cuff, virtually all of the vaccine can be in areas deeper than that Appropriate setting of the instrument allows a wide range of variation with regard to of these volumes

The measles vaccine can be any of those known to be useful in humans, e.g. the Endere B-level diluted live measles vaccine grown in chick embryo cell cultures. Such a vaccine is commercially available from Merck & Co., Ine, under the trade name RtJBEOVAX. Another useful measles vaccine is the Schwarz strain of diluted live virus, which is also commercially available. The measles vaccine used should be infectious such that a single human dose of the final product contains at least 1000 TCID (J _{0 of} the diluted measles virus. (Currently the Division of Biologios Standards requires 100 TOID This may change). 109 813/1595

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If a commercial measles vaccine is not desired, the vaccine is used according to conventional methods manufactured. According to a representative method, a virus is used which is found in cell culture preparations from Chicken embryos are grown »The Eell cultures are grown in a synthetic nutrient medium, for example that referred to as Medium 199, the binder serum and appropriate ones Contains antibiotics. A pure culture is used as measles virus to infect the cell cultures of the Edmon8ton strain or any other acceptable measles virus strain that has been shown to be free from foreign viruses The virus obtained from the cell cultures is filtered through clarified to remove foreign cell waste. The virus preparation is then administered according to the Division of Biologic Standards of the National Institutes of Health, U.S. Public Health Service, checked. A vaccine, which conforms to all standard rules is acceptable for human use.

The smallpox vaccine can be any of those used in the canteen be recognized as suitable. It can be in the EaIb, in chicken embryos, in cell cultures of chick embryos or other acceptable tissue or by any other suitable technique propagated

Methods of producing and control testing of smallpox

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Vaccine are in the "Minimum Requirements" for smallpox vaccine (published by the Division of Biologi.ce Standards, the National Institutes of Health, U.S. Public Health Service ·). Most of the time the Smallpox vaccine from Kuhpoeken ("cowpox") - live viruses, those from the vesicles in the ect. derm of the calf or one Another suitable animal at the peak of the blistering resulting from the artificial infection of the animal with a suitable cowpox virus seed preparation. The vesicles become the calf or another Animal removed in the form of a pulp or as a "lymph" and tested according to the regulations of the Division of Biologies Standards of the National Institutes of Health. The pulp is then treated by a process in which foreign protein is removed and bacterial contamination and PiXze noticeably reduced or completely eliminated will. Then the pulp tests for safety, purity and subjected to effectiveness.

The two vaccines will be a suitable stabilizer added to a loss of effectiveness during the Avoid drying or keep it to a minimum. The final product can also be neomyoin sulfate or Contain another acceptable antibiotic or preservative.

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The mixing of the two vaccines is done under aseptic conditions. The mixed vaccine is standardized so that the dose volume of the mixture intended for human use contains at least 10 * TCID ₅₀ (50 # tissue culture ansteocating dose) of measles virus and about 10 spotting units (PPU) per 0.1 ml of cowpox virus. The virus titre obtained by the stain method is usually equal to that obtained by observation of the oytopathic effect (TGXDe ₀ ). These levels of virus usually elicit antibodies to both types of virus in virtually all nonimmune individuals. Practically all individuals also have a primary smallpox infection ("take") with scars that appear »

The combined vaccine is bottled in glass containers, frozen at -60 ⁰ C and then dried by lyophilization.

The following examples describe the preparation of various representative combined vaccines.

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example 1

A measles vaccine with an ⁿ pre-pool ^w titer of 10 ^ ** TCIDkq / 0.1 ml was used.

The Euhpoxvirus had a calculated titer of 10 "PPu / ml. The Pooken vaccine was used with the measles vaccine mixed in a ratio of 1:59 parts

The combined vaccine was filled in portions of 11 ml, ^{frozen at -60 0} G and dried by lyophilization. For testing or use, the ampoule was refilled with U ml of sterile, distilled water.

The analysis of this mixture showed the following results: cowpox fleoken analysis: 10 ^ '/ 0.1 ml cow pox pipe analysisϊ 10 ^f TOIDcq / 0.1 ml measles analysis: 10 ^2f9 TCIDkq / 0.1 ml.

Examples 2 to 5

The following examples illustrate variations of the Example 1 with regard to the dilution of the smallpox vaccine in the rabbit vaccine.

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Example 2: 1 part smallpox - 9 parts measles

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These mixtures showed the following analysis results;

Smallpox vaccine Example Description TCID ₅₀ / PFÜ / 0.1 ml TCID ₅₀ /

0.1 ml 0.1 ml

2 frozen
dried 10 ^{5 6}
10 ^{5 6} ίο ^{5 9} I.
10 ^{2 9 5} frozen
dried 10 + ⁷ 10 ^{5 1}
10 ^{5 1} 10 ^{2 8} 4th frozen
dried io ^{5 6} Se 10 ^{2 6} 5 frozen
dried 10 ^{2 6}
io ² * ίο ^{3 0} ΙΟ ^{2 9}

Example 6

In the same way as in example 1, only with the modification, that different smallpox and measles vaccine amounts

9920

a mixture was made.

virus

Examination results TCID ₅₀ / O, 1 ml PFtJ / 0.1 ml

Smallpox, frozen XO ⁺ , 7 Smallpox, dried .1O ⁴ .6 Hasern 10 ² , 7

ΙΟ ⁵ ' ² 10 ⁵ · ¹

Virtually all of the disease-prone children who so far with the combined vaccine of the example were vaccinated, showed a primary Pookenimpfafcof f-

Infection ("take"), developed blood cell agglutinates

nation-inhibiting antibodies against cow pooken virus and were immune to smallpox, as evidenced by the excitement with smallpox vaccine, at a later date after the Using the usual multiple puncture method, Praktisoh showed that all children responded serologically to the measles vaccine and developed antibodies with titer levels approaching those values which obtained when measles virus vaccine is administered alone will.

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These examples illustrate that mixtures of the two live virus vaccines are compatible and stable and a dried product result in »the sun immunize of humans is highly effective

The combined vaccines should have a minimum amount of

10 * SOID ₅₀ of measles virus and a minimum age of

2 0
Contains 10 * PPU / 0.1 ml of cowpox virus per dose.

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Claims (3)

Merck "Co., Inc. 9920 (M 70 785) claims aaaaaaaaaaaaaaaäaaaaaaaaaaaaa 1. A method for producing a storage-stable, combined vaccine against Hasern and Poeken, characterized in that that one mixes rabbit and smallpox vaccine and then lyophilizes it. 2 * Method according to claim I _9, characterized in that the viruses are combined in a V ^ rhälfeni © of at least ^{10 5} TCID ^ ₀ units per dose of MaserwireR and ICr stain-forming units per 0.1 ml & ώ Kuhpo @ kemrir @ K. 3. The method according to claim I _3, characterized in that the viruses in a ratio of at least IG ¹ · ⁰ TCID, '50 kei the units per 0.1 ml of cowpox virus combined per dose in humans of measles viruses and 1O ^C »stain · Neuo documents (Art.?: 1, - ^ r :: * ■: II-109813/1598