JP2008540502A - Pharmaceutical composition for the treatment of acquired immune deficiency - Google Patents

Abstract

Use of smallpox vaccine virus and pharmaceutical composition for the preparation of a pharmaceutical composition for use in the treatment of infections resulting from acquired immunodeficiency.
[Selection figure] None

Description

  The present invention relates to a preparation for the treatment of acquired immunodeficiency syndrome (AIDS).

  To date, many researchers, including the applicant, have conducted research on the phenomenon of viral interference, for example, by studying the progress of anti-polio vaccines from serological and virological viewpoints.

  According to a study conducted by the applicants (at the time of the 1960s), due to the presence of numerous enteroviruses in the intestine at the time of vaccination, some patients vaccinated with polio vaccine lack an antibody response, The research hypothesis that immune defense defects would be observed was supported.

  On the other hand, this phenomenon does not occur when different types of viruses such as adenovirus are present in the intestine.

  These studies demonstrate the specificity of the virus interference phenomenon and that it occurs only in the presence of enteroviruses.

  This result, in turn, provided another important scientific confirmation that this viral interference is in the suppression of cell replication of viruses that have entered in the presence of enteroviruses.

  In the field of research on the effective treatment of acquired immune deficiency, the phenomenon of viral interference has been substantially underestimated, but the applicant It has been found that if the vaccine virus is attenuated appropriately, it can be effectively used as a carrier for immunization against the HIV virus causing disease.

  The object of the present invention is a therapeutic method used for the treatment and prevention of AIDS based on the virus interference phenomenon between HIV virus and smallpox vaccine virus when added in the presence of enzyme.

  In particular, the object of the present invention is to provide a medicament comprising an attenuated smallpox vaccine virus in the presence of an enzyme, preferably in the presence of bromelain, seratio-peptidase and endonuclease, preferably of the Elstree strain. Is the preparation.

  In the course of investigating the efficiency of therapeutic methods and related drug preparations, methods have been developed to perform both positive and negative specific viral interference phenomena in vitro.

  This method is a development of a basic method known as the “plaque method” that several researchers, such as Dr. Dulbecco, have already studied in the past.

In the following examples of the present invention, Elstry strain vaccine virus was used as a starting material. The test method includes the following steps:
Using continuous cell line SIRC cells trypsinized in EARLE growth medium and distributed in Kolle flasks (surface area 120 mm x 42 mm) divided into millimeter and part micrometer units, cell matrix The step of preparing
-Preparing a medium having the following growth medium composition:
First step - distilled water 90%, Earl solution 10 × 10% lactalbumin hydrolyzate enzyme 0.5 g%, 0.05% yeast extract, glutamine 0.015 g%, NaHCO ₃ 0.14 grams% Fetal calf serum 4%, calf serum 4%, penicillin 30,000U. I. And streptomycin 0.03 grams%;
Second Step-In the second test medium, only penicillin and streptomycin are each 500,000 U.S. I. And change to 0.5 grams%;
- it was added a solution 5% of distilled water and 10% NaHCO _3, with a mixture of Earle Medium purified agar and fold concentration of 1.8%, preparing a nutrient medium solids.

  In the method of the present invention, when the cell carpet is completed, the growth medium is removed, the culture is washed with a serum-free medium, and a small amount (0.2 cc.) Of appropriately diluted virus is overlaid on the cultured cells. Seeded.

  The virus suspension is brought into contact with the cell carpet at 37 ° C. for 45 minutes, taking care that the entire carpet surface is uniformly covered with liquid.

  Unabsorbed liquid is removed and the cell carpet is covered with 12 cc solid nutrient medium.

  When the agar has solidified sufficiently, the culture is inverted and allowed to stand at 37 ° C. After 5-6 days, plaques appear after a maximum of 8 days. At the time of observation, one of the colorants for viability determination, preferably trypan blue is added.

  The culture is allowed to stand for several days without any change to the necrotic area. This is because, as is clear from the observation results, the selected colorant binds to the viral DNA by inactivation in the presence of light.

  In terms of morphology, plaques formed by the vaccine virus were 2-4 mm in diameter 6 days after addition of the vaccine virus from microscopic observation.

  Subsequently, when appropriately diluted HIV virus was injected into the medium thus prepared, more extensive and widespread necrosis was observed.

  According to the present invention, endonuclease enzyme, which is a restriction enzyme, and two suspensions of bromelain and streptopeptidase as other enzymes were added to the medium.

  When appropriately diluted smallpox virus and HIV virus were added to the medium, no cytopathic plaque formation was observed under the same conditions as described above, but instead a marked reduction in toxicity was seen.

Claims (11)

  Use of smallpox vaccine virus for the preparation of a pharmaceutical composition for use in the treatment of infection resulting from acquired immunodeficiency, characterized in that said composition further comprises bromelain enzyme and seratio-peptidase enzyme Use of smallpox vaccine virus.   The use according to claim 1, wherein the smallpox vaccine virus is an Elstree strain.   Use according to claim 1 or 2, wherein the composition comprises a restriction enzyme.   4. Use according to claim 3, wherein the restriction enzyme is an endonuclease enzyme.   A pharmaceutical composition for the treatment of acquired immunodeficiency, comprising smallpox vaccine virus, bromelain enzyme and seratiothiopeptidase enzyme.   6. The composition of claim 5, comprising a restriction enzyme.   The composition according to claim 6, wherein the restriction enzyme is an endonuclease enzyme.   The composition according to any one of claims 5 to 7, wherein the vaccine virus is a diluted preparation. Furthermore distilled water, lactalbumin hydrolyzate enzyme, yeast extract, glutamine, NaHCO _3, fetal calf serum, calf serum, penicillin and streptomycin, composition according to any of claims 5-7.   A method for modifying a smallpox vaccine virus, comprising the step of culturing the virus in the presence of a bromelain enzyme and a serratio peptidase enzyme.   A vaccine virus modified by the method of claim 10.