WO1995035113A1 - USE OF FERROUS (III) OXYDE (Fe2O3) FOR THE TREATMENT OF IMMUNE DEFICIENCIES ESPECIALLY AIDS - Google Patents

Abstract

The invention relates to the use of very finely divided ferrous (III) oxide (Fe2O3) and/or its hydroxide for the treatment of immune deficiencies, especially Acquired Immune Deficiency Syndrome (AIDS), or of immune deficiency caused by the Human Immunodeficiency Virus (HIV). This basic and active substance with its antiviral effect is non-tocic, its dosage is not critical and has no side-effects, and is preferably intended for use as an oral medicament after the establishment of an immunodeficient disease in humans and animals.

Description

Use of iron (IΙI) oxide (FE203) for immunodeficiency, especially AIDS.

The invention relates to the use of iron (III) oxide in the case of immune deficiency, in particular aquired immune deficiency syndrome (AIDS) or against immune deficiency caused by the human immunodeficiency virus (HIV).

The drug zidovodine (3'-azido-3'-deoxithymidine) is known, but it can only temporarily delay the course of the disease; curative treatment must be ruled out. The tolerance of this preparation, which is also known under the name AZT, cannot be assumed for every patient. EP 0307914 uses a large number of different components in various combinations for the production of tablets, it being impossible to see which component is only approximately able to reinstate the body's own control mechanisms for bringing about a comprehensive cell resistance to the cell Virus or the bacillus causes. This requirement cannot be met by the application of iron-bound proteins such as actoferrin, transferrin and ovatransferrin, as described in EP 584558. The use of doped or modified iron oxide for therapeutic purposes (EP 0516252), e.g. in chemotherapy at best causes stress, especially in the case of pathophysiologically questionable conditions in a patient.

The object of the invention is to provide a preferably orally administrable, non-toxic medicament with an antiviral effect, which can be safely administered to humans and animals after detection of a cellular immune deficiency and the dosage is not a criterion. In addition, this fully tolerable and side effect-free drug should be used as a prophylaxis against most diseases caused by viruses. Serological examinations have pien with iron (III) oxide with the exclusion of any side effects led to the following results:

Virus detection (PCR) HIV-I negative; Antigen detection HIV-I negative; Antibody positive.

A rapidly developing product from an iron compound as a result of absorption of orally administered iron (IΙI) oxide, e.g. Hemoglobin leads to the accumulation of the molecular cell structure in the organism without having to fear stressful connections with the stomach acid when taking this basic / active substance. Because of this behavior because of the insolubility of iron (III) oxide towards acids, the administration of this compound does not conflict with the physique.

This object is achieved by the use according to the application of the iron (IΙI) oxide which is approved as a dye in foodstuffs as an active ingredient.

The antiviral activity of the solid iron (III) oxide against the AIDS pathogens HIV-1 and HIV-2 was found. This is illustrated by the test results below.

Since phytohemagglutinin-activated lymphocytes from the umbilical cord blood of newborns are more susceptible to HIV infection than those from the peripheral blood of adult donors-activated lymphocytes (PBL), the examiner is offered a more sensitive system. Therefore, in addition to the PBL, umbilical cord cells were used for the examination.

1. Examination in 24-hole plates

The iron (III) oxide was sterilized by heating in an autoclave (134 ° C., 60 min.) And suspended in culture medium after cooling. Aliquots of this suspension were mixed with phytohemagglutinin-activated lymphocytes from peripheral blood or from umbilical cord blood (final concentration 500,000 cells per ml) and incubated for 24 hours in an incubator at 37 ° C / 6% CO2. The cultures were then infected with the various HIV-1 and HIV-2 isolates shown in Table 1.

Table 1: Characterization of the virus isolates used

Code HIV subtype reverse transcriptase titer / ml (still

Activity (cpm / ml / 90 min. Visible CPE)

K31 HIV-1 276,000 10 ⁵ - 10 ⁶

D34 HIV-1 209,000 10 ⁵ - 10 ⁶

D194 HIV-2 470,000 10 ³ - 10 ⁴

LAV-2 HIV-2 156,000 10 ³ - 10 ⁴

2. Checks

Positive controls: experimentation without iron (III) oxide with activated umbilical cord ymphocytes or PBL, which were infected with the various HIV-1 and HIV-2 isolates.

Negative controls: experimentation with uninfected cells or their culture supernatants.

The cultures were assessed daily in a light microscope (examination for the development of a cytopathic effect (CPE) and by measurement of the retrovirus-specific enzyme reverse transcriptase (RT).

evaluation

Table 2 summarizes the light microscopic assessment of the cultures from three experiments carried out in parallel with PBL and one experiment with umbilical cord ymophocytes. They mean: - no visible cytopathic effect + visible cytopathic effect

Table 2: Assessment of cell cultures for the formation of syncytia (CPE) on day 4 with activated umbilical cord lymphocytes or on day 6 with activated PBL.

Iron (III) oxide virus isolate concentration (µg / ml) K31 D34 LAV-2 D194

20th

50 100 200

positive controls:

negative control:

In Table 2, the formation of syncytia could be prevented at all of the iron dll) oxide concentrations examined. An antiviral effect of iron dll) oxide on HIV-1 and HIV-2 could thus be clearly demonstrated. This result was achieved with both activated PBL and activated umbilical cord blood lymphocytes.

Eight days after infection of the activated cells, 1 ml of cell-free supernatant was removed from all cultures. The virus was pelleted in an ultracentrifuge and then subjected to a test for the retrovirus-specific enzyme reverse transcriptase (RT). The enzyme activity is a rough measure of the amount of virus in the supernatant of the cells, however not identical to the amount of infectious virus particles. It is determined by incorporating radioactive thymidine into the DNA. The results of this experiment are therefore given in cpm / ml culture supernatant per 90 minutes.

Table 3: Evaluation of the cell cultures (PBL) on the enzyme activity of the reverse transcriptase (cpm / ml per 90 min.) Eight days after the infection.

Iron (III) oxide virus isolate concentration (µg / ml) K31 D34 LAV-2 D194

20 517 526 196 580

50 570 277 426 208

100 325 489 400 560

200 581 693 298 701

positive

Control: 21734 6106 3699 41264

negative

Control: 573

It can be seen from Table 3 that the retrovirus-specific enzyme reverse transcriptase (RT) could practically no longer be detected in the culture supernatant at all dilution levels of the test substance. The enzyme activities are considerably below the value of the positive control.

A correspondence of this result with that of a light microscopic examination for the non-occurrence of cytopathic effects became clear.

The test results undoubtedly showed an anti-viral effectiveness of the iron (III) oxide against the under- departed! HIV strains.

In addition, the non-toxicity in the range of the concentrations used could be proven without any problems.

For ulation:

Iron (III) oxide in fine-grained form and / or its hydrate is used as the starting material for the production of suitable medicaments.

The iron (III) oxide can be prepared as such or in the form of uninvolved, non-toxic, solid or liquid extenders or carriers, which are in solid or liquid form, e.g. capsules, coated tablets, microcapsules, suppositories, suspensions, emulsions can be used.

The p.o.-usable preparations such as capsules or coated tablets are to be preferred. The production takes place in a manner known per se.

The dose of iron (III) oxide to be administered largely depends on the field of application and the type of formulation. Included in the following subsequent examples

1. a preferably stomach-soluble gelatin capsule 400 milligrams of fine-grained iron (III) oxide with a grain size of approximately 5 micrometers, intended for daily therapy by oral application in the case of immunodeficiency or

2. The same active ingredient mentioned under 1. with a carrier substance is pressed into pills.

Claims

PATENT CLAIMS

1. Use of iron (III) oxide as an active ingredient for the treatment of immunodeficiency disorders, in particular "AIDS".

2. Use according to claim 1 of iron (III) oxide hydrates.