RU2257916C1 - Liquid preparation containing immunoglobulin efficient against marburg fever from horse blood serum (horse immunoglobulin marburg) - Google Patents

Abstract

FIELD: clinical immunology.

SUBSTANCE: preparation represents 9.0-11.0% solution of horse blood serum immunoglobulins with pH 7.0-7.5 and content of γ-immunoglobulin fraction at least 90.0% of the total protein. Nonspecific fractions in preparation are albumin, α- and β-globulin fractions in amount not larger than 10.0%, and residual ethyl alcohol in amount not larger than 4.0%. Preparation is characterized by virus-neutralizing antibodies against virus Marburg. Titer of virus-neutralizing antibodies is at least 1:2048 and can be used for emergency prevention of Marburg fever in humans.

EFFECT: extended immunoglobulin application area.

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Description

The invention relates to medicine, specifically to drugs and medicines (to immunoglobulins) for therapeutic purposes, and can be used for emergency prevention of Marburg fever in humans.

Specific means of prevention and treatment of Marburg fever, which is a severe febrile illness occurring with a pronounced hemorrhagic syndrome and damage to almost all organs with a mortality rate of up to 80% [1], are currently lacking. Treatment of patients is symptomatic.

It has been shown that with Marburg fever, the use of serum convalescents is effective [2]. However, our country does not have its reserves. In addition, the disadvantage of serum is that it can be contaminated with immunodeficiency viruses and hepatitis B. The use of such a serum is associated with the risk of infection of people with infectious pathogens.

There is data on the development of therapeutic and prophylactic preparations of immunoglobulin against Marburg fever from the immune serum of sheep and goats, which are 12% solutions of immunoglobulins isolated by the alcohol method in the cold from hyperimmune serum of goats or sheep (prototype) [3, 4]. The preparations contain gamma-globulin fraction and impurities in an amount of not more than 10.0% of the total protein. If used for emergency prevention of experimental Marburg fever in animals, they showed protective properties.

The disadvantage of sheep and goats selected as a producer of immune serum is that they do not provide the necessary amount of immune serum, and their serum may be contaminated by foreign microorganisms (for example, scrapie in sheep). They can also be carriers of spongiform encephalopathy [5], and immunoglobulin preparations isolated from the blood serum of these animals can be a source of infection of humans with infectious agents when they are introduced. Attempts have already been made to use sheep, goats, mules, donkeys and cattle to produce hyperimmune sera. However, these studies did not bring the desired results.

In addition, preparations of immunoglobulin against Marburg fever from the blood serum of sheep and goats are at the stage of development and study of the properties of experimental samples, and data on their effectiveness [3, 4] do not allow us to unambiguously evaluate their protective properties and the possibility of using drugs for the prevention of Marburg fever in people in case of their specific defeat.

It was previously noted that, in addition to sheep and goats, horses also respond by the production of specific antibodies in response to their infection with the Marburg virus [4, 6, 7]. Horses have high immunological reactivity and provide large volumes of serum. Therefore, horses are most often used to obtain specific hyperimmune serums, which are the feedstock for the isolation of immunoglobulins [8].

The blood serum of horses is free from the deficiencies inherent in the blood serum of sheep and goats. Currently, horses remain a recognized producer of hyperimmune sera and they are mainly used on an industrial scale. In addition, there is experience in obtaining immunoglobulin preparations against a number of viral infections from horse serum [8-10]. Based on this, a horse was selected as the producer of the hyperimmune serum for the Marburg virus.

The aim of this invention is to develop a drug containing immunoglobulin against Marburg fever from blood serum of equine horses, intended for emergency prevention of Marburg fever in humans.

This goal is achieved due to the ability of the Marburg virus to induce the production of specific protective antibodies in animals in response to infection, and the use of native hyperimmune blood serum of horses containing antibodies to the Marburg virus as a raw material for the preparation.

The essence of the invention lies in the fact that the preparation containing immunoglobulin against Marburg fever liquid, is a 9.0-11.0% solution of immunoglobulins of hyperimmune blood serum of horses containing antibodies to the Marburg virus, has a neutralizing activity against the Marburg virus and with the introduction It has a pronounced preventive effect in people with Marburg fever.

Execution example

The raw material for the preparation of an immunoglobulin preparation is hyperimmune horse serum containing antibodies to the Marburg virus. Isolation of immunoglobulin from the blood serum of horses containing antibodies to the Marburg virus is carried out by the method of alcohol fractionation in the cold according to Cohn [11].

A preparation containing immunoglobulin against Marburg fever from horse serum is a clear or slightly opalescent liquid, colorless or with a yellowish tinge, sometimes with a slight precipitate that breaks easily when shaken, with a pH value of 7.0 to 7.5 and is 9.0-11.0% solution of horse serum immunoglobulins containing antibodies to the Marburg virus. The content of the gamma globulin fraction in it is at least 90.0% of the total protein. Albumin, α- and β-globulins in the amount of not more than 10.0% and residual ethyl alcohol, the content of which is not more than 4.0%, are present as non-specific impurities in the preparation. The drug has a neutralizing activity against the Marburg virus, characterized by a titer of neutralizing antibodies of at least 1: 2048.

The drug is sterile, pyrogen-free and non-toxic. With intramuscular administration, it has a pronounced prophylactic effect with Marburg fever.

According to physicochemical and immunobiological parameters, the equine immunoglobulin preparation Marburg corresponds to the quality indicators of commercial heterologous immunoglobulins isolated from horse serum [8] and is recommended for emergency prophylaxis of Marburg fever in humans.

9 series of an immunoglobulin preparation against Marburg fever were obtained from horse serum. The characteristics of the drugs are presented in the table.

The preparation containing immunoglobulin against Marburg fever from horse serum, liquid, (equine Marburg immunoglobulin) is certified by GISK im. L.A. Tarasevich and approved by the IIBP Committee. Regulatory and technical documentation for the drug is submitted for consideration to the Pharmacopoeia Committee of the Ministry of Health of the Russian Federation.

Table
Characterization of heterologous immunoglobulin preparations obtained from horse serum Name of quality indicator, unit of measure The value of the indicator for immunoglobulin preparations heterologous commercial [8] against Marburg fever (9 series data) Appearance Transparent or slightly opalescent liquid, colorless or with a yellowish tinge. A slight precipitate is allowed, easily breaking when shaken. Transparent or slightly opalescent liquid, colorless or with a yellowish tinge. A slight precipitate is allowed, easily breaking when shaken. The concentration of hydrogen ions, units pH 7.0 to 7.5 7.0 to 7.5 The amount of total protein, percentage 9.0 to 11.0 9.0 to 11.0 The content of gamma globulin fraction, percentage 70.0 to 95.0 [12] 90.0 to 97.4 The content of residual ethyl alcohol, percent No more than 4,5 1.4 to 3.7 Pyrogenicity Pyrogen-free Pyrogen-free Toxicity Non toxic Non toxic Sterility Sterile Sterile Expiration date, years No more than 3 3

BIBLIOGRAPHIC DATA

1. Muhlberger E., Weik M., Beeker S., Klenk H. D. The replication and transcription strategy of Marburg and Ebola viruses: differences and similarities // Infect. Dis. Rev. - 2000. - Vol.2, No. 4. - P.251.

2. Stille W., Bohle F., Helm E. et al. Uber eine durch cercopithecus aethiops ubertragene infektions-krankheit / "Grune Meerkatzenkrankheit", "Green Monkey Discase" // Dtsch. Med. Wschr. - 1968. - Bd. 93. - S.572-582.

3. Kudoyarova N. M., Kizimov N. V., Smolina M. P., Dedkova L. M. Study of the protective properties of gamma-globulin preparations against Ebola and Marburg viruses // Actual problems of biotechnology: Materials of the 2nd industry conference April 25-27, 1990, urban settlement Koltsovo, 1990 .-- P.53-54.

4. Kizimov N.V., Kudoyarova N.M., Kaliberov S.A. et al. Development of therapeutic and prophylactic preparations of gamma globulin against diseases of Marburg, Ebola, Bolivian hemorrhagic fever // Actual problems of medical biotechnology: Materials of a scientific conference dedicated to the 85th anniversary of the Tomsk Scientific Research Institute of Vaccines and Serums, NPO Virion, Tomsk, 1991. - Part 2. - S. 31-32.

5. Virology / Ed. B. Fields - M., 1989 .-- T.3. - S. 419-430.

6. Krasnyansky V.P., Pshenichnov V.A. Gonchar N.I. and other Immunological features of hemorrhagic fevers Marburg and Lass and the prospects for their specific prevention // First All-Union Immunological Congress, Sochi November 16-17, 1989: Abstracts of section and poster presentations. - M., 1989 .-- T.1. - S. 226.

7. Gonchar N.I., Pshenichnov V.A., Pokhodyaev V.A. and other Sensitivity to the Marburg virus of various experimental animals // Vopr. virusol. - 1991. - No. 5. - S. 435-437.

8. Guidelines for vaccine and serum administration / Ed. P.N. Burgasova. - M., Medicine, 1978.

9. Krasnyansky V.P., Gradoboev V.N., Borisevich I.V. et al. Development and study of the properties of immunoglobulin against Lass fever // Vopr. virusol. - 1997. - No. 2. - S. 71-74.

10. Borisevich I.V., Mikhailov V.V., Krasnyansky V.P. et al. Development and study of the properties of immunoglobulin against Ebola / Vopr. virusol. - 1995. - No. 6. - S.270-273.

11. Cohn E.J., Strong L., Hughes W.L. et al. Preparation and properties of serum and plasma proteins // J. Amer. Chem. Soc. - 1946. - Vol. 68, No. 3. - P.459-475.

12. Handbook of the use of bacterial and viral drugs / Ed. S.G. Dzagurova, F.F. Rezepova. - M., Medicine, 1975.

Claims (1)

Liquid drug containing immunoglobulin against Marburg fever from horse blood serum (horse immunoglobulin Marburg), which has neutralizing activity against Marburg virus and is used for emergency prevention of Marburg fever in humans, characterized in that hyperimmune horse blood serum is used as the starting material for its preparation and representing a 9.0-11.0% solution of immunoglobulins of hyperimmune blood serum of horses containing antibodies to the Marburg virus containing γ-globulins fraction in an amount of not less than 90.0% of the total protein content, containing non-specific impurities containing albumin, α- and β-globulin fractions in an amount of not more than 10.0% and residual ethyl alcohol in an amount of not more than 4.0%, having a pH value from 7.0 to 7.5 and characterized by a titer of neutralizing antibodies of at least 1: 2048.