RU2245178C1 - Method for acting upon aids virus - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves exposing cells containing virus to radiation. Pulsating monochromatic light with pulses succession frequency being equal to 9.4 Hz is applied with wavelength group of 516.7, 517.2, 518.2 nm and then with 636.2 nm of total power not less than 400W. Power ratios in wavelength group is first equal to 1:0 then 1:1,1:3, 3:1, 0:1. Exposure time is equal to at the level of 20 min.

EFFECT: enhanced effectiveness of treatment.

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Description

The invention relates to virology, to a means of radiation inactivation of the AIDS virus.

A known method of treating AIDS virus using the drug azidothymidine (1). This drug is able to block the synthesis of viral DNA by reverse transcriptase, i.e. the process when genetic information is transcribed not from DNA to RNA, but rather from RNA to DNA.

Azidothymidine is able to alleviate the course of the disease and prolong the life of AIDS patients. Disadvantages - high toxicity, especially for bone marrow, contributes to the development of anemia (a decrease in the number of red blood cells), and with its frequent use, the number of leukocytes and platelets can decrease.

In addition, this drug can partially block the life cycle of the AIDS virus only at one stage - viral DNA synthesis by reverse transcriptase.

The closest analogue of the proposed method is a method of exposure to the AIDS virus, which was irradiated with light with a wavelength of 636.2 nm with an exposure of 30-60 minutes and light corresponding to electronic transitions in magnesium atoms (516.7; 517.2; 518.2 nm) (RU 2097427, 11.27.1997).

The disadvantage of this method is the low inactivation of the AIDS virus.

The purpose of the invention is the simultaneous blocking of the AIDS virus at all its life stages.

The life cycle of HIV is a complex multi-step process. The first step of HIV, like any other viral infection, is the binding of HIV to the membrane of the infected cell.

The mechanism by which HIV enters the cell is that one of the proteins of the viral envelope, namely dr120, binds to the membrane protein of the infected cell, called the CD-4 antigen. Then the virus membranes and cells fuse, as a result of which viral RNA and reverse transcriptase penetrate the cell cytoplasm. Here, the viral genome is copied and, using reverse transcriptase, viral DNA is formed, which is integrated into the DNA of the host cell, thereby ensuring the synthesis of viral RNA and proteins from which new viruses are assembled. They leave the cell, violating the integrity of its membrane, and then infect other cells.

It follows that the virus that hit the cell is capable of changing it in such a way that it itself becomes a stranger to its own organism and is destroyed by its own immune system.

From the foregoing, we see what a complex path the virus takes until it forms new virions. By the way, from the point of view of scientific research, such a multi-stage life cycle of HIV is a positive factor, since there are many opportunities to intervene in this cycle and disrupt it, and this is precisely the treatment for AIDS.

It is clear that interruption of at least one life cycle will cause death or stop the multiplication of HIV. The very concept of treatment for the AIDS virus includes the creation of a drug that specifically violates certain stages of its life cycle.

Analyzing the life cycle of the virus, one cannot but pay attention to the fact that the genetic material of HIV plays a key role in this process. First, viral RNA serves as a matrix for the synthesis of viral DNA, then this proviral DNA again forms viral RNA to assemble new viral particles, etc.

An analysis of studies performed in recent years has shown that, as a rule, these studies are aimed at developing a drug that allows you to block the life cycle of the virus at any particular stage. In this regard, the stage at which proviral DNA with viral RNA is generated using reverse transcriptase is noteworthy.

However, in a human body infected with HIV, all life stages of HIV can be carried out simultaneously, from the introduction of HIV into the cell to the assembly and release of new viral particles from the cell. Therefore, a drug that blocks one stage (for example, DNA synthesis) may not be effective for another stage (for example, for the stage responsible for the assembly of new viruses).

It is clear that the greatest effect of treating the AIDS virus will be when all stages of the HIV life cycle are simultaneously blocked. This procedure can be carried out using light with a long wavelength corresponding to electronic transitions of magnesium and zinc atoms (magnesium is present in the RNA molecule, and zinc in the enzyme reverse transcriptase).

Moreover, irradiation should be carried out with lamps simultaneously filled with vapors of magnesium (wavelength 516.7; 517.2; 518.2 nm) and zinc (wavelength 636.2 nm), but in different ratios. These ratios must be such that the amount of light emitted from one lamp corresponds to a quite definite value, i.e., for example, a lamp filled with magnesium and zinc vapor having a power of 400 W should emit, for example, 100 W of light corresponding to magnesium, and 300 watts corresponding to zinc, etc.

Our studies showed that the greatest suppression of the virus located in the cell occurs if the irradiation is carried out with pulsed monochromatic light with a pulse repetition rate of 9.4 Hz simultaneously with a group of wavelengths 516.7; 517.2; 518.2 nm, and then with a wavelength of 636.2 nm, with a total power of at least 400 W with a ratio of the power of the group of wavelengths to the wavelength of 636, 2 nm, first 1: 0, then 1: 1, 1: 3, 3: 1, 0.1 for at least 20 minutes.

Such a range of illumination blocks almost all vital stages of HIV from its introduction into the cell to the assembly of new viruses. It is not necessary to know at what stage HIV is in the body.

Experiments have been performed with the goal of virucidal efficacy of monochromatic radiation sources for HIV. The experiments were carried out at the Institute of Virology. D.I. Ivanovskogo RAS.

Materials and methods.

Viruses. Used the strain of HIV - ISTOA from the collection of viruses of the Research Institute of Virology. Ivanovo RAMS.

Cells. Used transplantable human lymphoblastoid cells MT-4 from the cell collection of the Research Institute of Virology. D.I. Ivanovo RAMS. Cells were cultured at a concentration of 3-5 × 10 in 1 ml in RPMI 1640 medium with 10% serum of cow embryos (Sigma, USA), 100 μg / gentamicin (Pharmachim, Bulgaria). Cell viability was determined by staining with a 0.4% trypan blue solution (Serva, Germany).

Two 24-well plastic panels (Costar, USA), in which a cell suspension was placed, were taken for investigation. Then the suspension in the panels was infected with HIV and cultured at 37 ° in an atmosphere of 5% CO ₂ 98% for 5-9 days. Moreover, one panel after placing the virus in it was irradiated with light, and the second (control) was not irradiated. Used lamps with a power of 400 and 450 watts. Reproduction of HIV in sensitive cells was assessed by their viability by the virus-induced cytopathic effect (CPE) and syncytia (giant multinucleated cells).

The results are presented in the table, from which it follows that irradiation of HIV with monochromatic light significantly reduces the infectivity of the test virus-HIV. Syncytia disappeared in all variants of virus dilution. This effect was most pronounced after 9 days of observation. Thus, the experiment showed the effectiveness of the proposed method for the treatment of the AIDS virus.

LITERATURE

1. Robert Yarkoyan, Hiroaki Mitsuya, Samuel Broder. AIDS treatments. In the world of science No. 12, 1988, 78-88.

Table The effects of monochromatic light on human immunodeficiency virus Breeding HIV infection Light exposure The control 1* 2 3 1 2 3 1: 2 67 0.7 0 46 0.8 60 1: 4 81 0.9 0 56 0.5 40 1: 8 81 1.3 0 56 0.7 36 1:16 83 1.8 0 61 0.7 32 1:32 88 1.4 0 82 0.9 25 1:64 87 1,6 0 84 1.3 21 1: 128 85 1.3 0 90 1,6 16 1: 256 86 2.0 0 92 1.3 7 1: 512 90 1,6 0 89 1,5 6 1: 1024 96 1.9 0 89 1,6 0 Experience Conditions 1 2 3 The control 96 1.9 0 Note:
1 * - cell viability,%
2 ** - number of cells × 10 ⁶ / ml
3 *** - syncytia, quantity / well

Claims (2)

1. The method of exposure to the AIDS virus, including irradiating the cells containing the virus with light, characterized in that the irradiation is carried out with pulsed monochrome light with a pulse repetition rate of 9.4 Hz simultaneously with a group of wavelengths 516.7; 517.2; 518.2 nm, and then with a wavelength of 636.2 nm, with a total power of at least 400 W with a ratio of the power of the group of wavelengths and wavelength of 636.2 nm, first 1: 0, then 1: 1, 1: 3, 3: 1, 0: 1. 2. The method according to claim 1, characterized in that the irradiation is carried out for at least 20 minutes