LIVE ATTENUATED CULTURE VACCINE FOR PREVENTION OF SMALLPOX AND OTHER ORTHOPOXVIRUS INFECTIONS BASED ON SMALLPOX VACCINE VIRUS AND METHODS FOR ITS PREPARATION AND APPLICATION Russian patent published in 2022 - IPC C12N15/00 C12N7/00 A61K39/285 

FIELD: biotechnology.

SUBSTANCE: presented vaccine contains a strain VACΔ6 of the smallpox vaccine virus with six disrupted virulence genes C3L, N1L, J2R, A35R, A56R, B8R and a stabilizer including sucrose, gelatin, peptone from soy and sodium chloride with the following content of these components of the finished form of the drug after freeze drying, wt.%: peptone from soy - 30.3, sucrose - 30.3, gelatin - 30.3, a mixture of lyophilizate of the recombinant strain VACΔ6 of the smallpox vaccine virus with a titer of 10⁷ OOE and sodium chloride - 9.1. The method for obtaining the vaccine involves infecting the monolayer of cell culture with the seed material of the attenuated smallpox vaccine virus - recombinant strain VACΔ6 of the smallpox vaccine virus with six disrupted virulence genes, incubation of the virus in a cell monolayer in a serum-free nutrient medium at a temperature of 37°C until 100% cytopathic effect is obtained, freezing of the resulting virus-containing suspension at the stage of its collection and accumulation, purification and concentration of the resulting viral suspension by ultrasound treatment and centrifugation, introduction into purified virus-containing suspension of stabilizing composition in the form of aqueous solutions of sodium chloride, peptone from soy, sucrose and gelatin and lyophilic drying of the resulting mixture of these components to obtain a vaccine in dry form. As a cell culture for the development of the virus, a digested cell culture 4647 of the African Chlorocebus monkey kidney is used, and as a serum-free nutrient medium, a DMEM nutrient medium containing an additional 10 units /ml of endonuclease benzonase and 1.5 mm MgCl₂ is used. A sterile infusion saline solution of NaCl and components of a stabilizing composition are injected into the purified virus-containing suspension at the rate of 27.6 ml of a 25% aqueous gelatin solution, 27.6 ml of a 25% aqueous peptone solution from soy, 13.7 ml of a 50% aqueous sucrose solution per 160 ml of an infusion saline solution of sodium chloride to obtain a titer of a virus-containing suspension of at least 5×10⁷/ml with the following content of these vaccine components before freeze drying: peptone - 0.006 g / 0.2 ml; sucrose - 0.006 g / 0.2 ml; gelatin - 0.006 g / 0.2 ml; suspension of the recombinant strain VACΔ6 of the smallpox vaccine virus with a titer of about 10⁸ OOE in an infusion saline solution of sodium chloride - the rest is up to 0.2 ml. The method for application of the vaccine is also presented. It is characterized by the fact that a vaccine obtained on the basis of a recombinant strain of the VAC-6 smallpox vaccine virus with six disrupted virulence genes is used for vaccination, which is prepared as follows: with a single vaccination scheme, one dose of a dry drug with a titer of 10⁷ OOE is dissolved in 0.2 ml of water for injection or sodium chloride solution for infusions and fill an insulin syringe in a volume of 0.2 ml with a titer of 10⁷ OOE, and with a double vaccination scheme, the contents of the ampoule - 10 doses of a dry drug with a titer of 10⁷ OOE are dissolved in 2.0 ml of water for injection or sodium chloride solution for infusions and an insulin syringe is filled in a volume of 0.2 ml with a titer of 10⁶ OOE. The drug is administered with a single vaccination intradermally at a dose of 1×10⁷ ООЕ, and with a double vaccination - intradermally at a dose of 1×10⁶ ООЕ with an interval of 28 days.

EFFECT: expansion of the nomenclature of live highly attenuated and safe vaccines against smallpox and other human orthopoxvirus infections

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