HUT73382A - Drogs for the treatment of retrovirus infections - Google Patents

Description

The present invention relates to pharmaceutical compositions for treating retroviral infections.

Retroviruses are spherical, enveloped RNA viruses of which two subfamilies are pathogenic to humans and animals. Retroviruses are characterized by the presence of reverse transcriptase, an RNA-dependent DNA polymerase. During replication, single-stranded viral RNA is transcribed by reverse transcriptase into double-stranded intermediate DNA and incorporated into the host cell genome as a provirus. By regular transcription from the provirus, the formation of RNA-containing viral particles, which emanate from the host cell, begins. Retroviruses can cause leukemia, autoimmune diseases, and immunodepression, and in particular, in humans, AIDS and ARS, or AIDS Related Syndrome, a precursor to advanced AIDS disease. So far, it has not been possible to cure retroviral infections in humans because previous attempts to develop vaccines against the virus have failed due to the high variability of the virus. To date, drugs that inhibit reverse transcriptase, the most commonly substituted guanosine, cytidine and thymidir, have been used in the treatment. dezoxinukleozidokat. The most well-known active ingredient so far is AZT, which can slow down retroviral infection but cannot cure the disease.

Surprisingly, we have found that bacterial and protein-free extracts from Escherichia coli, optionally in combination with extracts from Gingko biloba,

retroviral infections would be significantly slowed down without any indication of side effects of nucleoside preparations. The coli extracts can be used alone with the gingko extracts, but it is also possible to combine the extracts with the treatment with the nucleoside compositions to reduce the dose of the nucleoside and thereby reduce the expected side effects.

Gingko biloba leaf extract has long been used to treat peripheral and cerebral circulatory disorders. The preparation of extracts from green Gingko leaves is described, for example, in DE PS-17 67 098, DE-PS 21 17 429, EP-A 0 324 197, EP-A 0 330 567 and DE-OS 39 40 091. Another method for extracting yellow autumn leaves of Gingko biloba from EP-B 0 352 146 is known.

The leaves of Gingko biloba contain many different compounds, the predominant of which are the so-called gingko flavon glycosides. A typical representative of these compounds is 5,7,3 ', 4'-tetrahydroxyflavono-3-O-α-rhamnopyranosyl-4-O-B-D- (6'-transcoumaroyl) glycopyranoside. Small amounts of terpene lactones are also present, such as gingcolides, such as those described, for example, by Okabe et al., J. Chem. Soc. (1967) 2201-2206. Bilobalide is a close relative of terpene lactones, the use of which in the treatment of neuropathy and the like is known from U.S. Pat. No. 4,571,407.

In addition, Gingko biloba leaves are called gingkol-

they also contain acids which are 6-alkyl salicylic acid derivatives having 13 to 19 carbon atoms and 0-3 double bonds. From these gigolic acids, the corresponding known phenol can be obtained biogenetically and by processing the leaves. The process for preparing the extracts differs in essence from attempting to eliminate as much as possible unwanted accompanying components, especially when the extracts are to be used in injectable form. Many methods of removing unwanted components are known to those skilled in the art. Accordingly, commercially available gingko extracts are well tolerated in injectable form. Such products are, for example, marketed under the tradename Tebonin by Dr. Wilmar Schwabe GmbH & Co. (Karlsruhe).

EP-B 0 352 146 also mentions that the extract of yellow leaves of Gingko biloba can reduce high levels of gamma-globulin in mammals, such as those observed in AIDS patients. However, this document does not elaborate on the success of the extract in AIDS.

Bacterial and protein-free filtrates of cultures of Escherichia coli cultures have long been used in medicine, partly for motility and permeability disorders of the intestinal mucosa and for inflammation such as Morbus Crohn or natibiotic, chemo or radiation therapy. Another application of coli extracts is in the treatment of allergies such as hay fever, urticaria, eczema, food allergy and asthma. Finally, an additional area of application is the treatment of acne and migraine, and therefore these extracts, such as those described in DE-PS 38 16 298, are anabolically effective and stimulate non-pathogen-specific immune defenses. The production process is described, inter alia, in DE-PS 38 16 298. Bacterial and protein-free extracts from Escherichia coli from relatively short oligopeptides and, as in the Drug Slit. 23, 829-830 (1973). Nucleic acids could not be detected. It is striking that quantitative analysis of peptides confirmed the absence of aromatic amino acids other than histidine.

Such cell-and protein-free extracts of Escherichia coli may be administered orally or by injection. Suitable formulations are, for example, those marketed under the tradename Colibiogen by Laves Arzneimittel GmbH.

The effect of coli extracts on retroviral infections, especially when combined with gingko extracts, cannot yet be scientifically explained. However, clinical trials clearly show that such treatments improve the general condition of patients and that the proportion of T ₄ lymphocytes remains constant, and in most cases increases. According to a recently developed working hypothesis, the specific (2 ·> - · «······ tt ο ······· ♦ ·· immune defense observed loss assumed based on that of retrovirus infection of T ₄ and -lymphocytes subsequently causes a reduction ₈ -lymphocytes of T. the production of non-directed immunoglobulins whereupon by B lymphocytes excessive amount leads to a specific immune defense impairment, and then collapse. Clinical observations is believed that coli and ginkgo active ingredients in extracts reduce or prevent increased production of non-directed immunoglobulins.

It has also been found that the dose of retrovirus-specific drugs can be reduced during the administration of coli and gingko extracts, thereby significantly reducing the risk of side effects, especially secondary anemia. The dose reduction seems to be only partially justified by the fact that these extracts can be used with nucleoside formulations. It is not yet clear to what extent the combination is compatible with other biochemical mechanisms.

The pharmaceutical compositions of the invention may be administered orally, but preferably by injection. It has been found that the coli and gingko extracts can be coadministered with the nucleoside formulations, and it is preferred that the coli and gingko extracts are administered prior to the nucleoside formulations, the interval between administration of the two formulations being from a few hours to 1-2 days. range. The invention therefore also relates to a kit comprising the individual active compounds in a spatially separated form for oral or parenteral administration.

The dosage unit for coli extract is 4-9 x 10 ¹² protein-free and cell-free metabolites from chickens, while the dosage unit for gingko extract is about 50-200 mg dry matter extract. The daily dose should be individualized for each patient. No long-term side effects have been reported so far.

The invention is illustrated by the following non-limiting examples.

Example 1

A coli extract was prepared as described in DE-PS 38 16 298 by inoculating Bouillon's medium with the desired serotype E.coli strain and cultivating it at 37 for at least 5 days. Preferably, the serotype O2: K1: K6 is used. The resulting Bouillon culture at a concentration of about 4-9 x 10 colony forming units / ml is prepared in a known manner under bacterial and protein-free filtration under sterile conditions, which is either processed into a solution for injection or used as an oral preparation alone. In addition, the extracts are freed of water by gentle vacuum drying and converted into capsules or tablets in known manner.

Example 2

Gingko extract is prepared by finely chopping green or yellow leaves of Gingko biloba and extracting it with aqueous acetone or ethanol, in particular water / acetone / ethanol (50:50 v / v) for several hours at 40 to 60 ° C. After extraction, the liquid from the leaves is squeezed out and the leaves are extracted once more. The resulting aqueous extract is extracted in a first purification step with a water immiscible solvent, preferably cyclohexane. The aqueous-ketone phase is then worked up and the non-polar phase discarded. The aqueous ketone phase is concentrated to about half its volume and the precipitate formed is filtered off. The filtered solution is treated with ammonium sulfate in a known manner and extracted with butanol. The resulting extract was dried over sodium sulfate under reduced pressure and then extracted extensively with ethanol. The residue was evaporated to dryness in vacuo.

The resulting extract, which can be formulated in a conventional manner, contains on average about 25% gingcoflavon glycoside and about 5% terpene lactone, which can vary by + 10% from the origin of the leaves.

Example 3

Clinical trials to date indicate that oral administration of one teaspoon or two days of coli extract orally to ARS patients results in a significant improvement in general condition and an increase in the T ₄ lymphocyte population. Addition of an ampoule of gingko extract containing 50 mg of dry matter in addition to the coli extract, even in severe cases, increases the T ₄ lymphocyte count above 400 / μl. The coli and gingko extract administered with nucleoside treatment results in a significant reduction of the subjective and objective side effects, thus reducing the dose for longer treatment.

Claims (7)

Claims CLAIMS 1. A pharmaceutical composition for treating a retroviral infection comprising a bacterial and protein-free extract of Escherichia coli known per se. Pharmaceutical composition according to claim 1, characterized in that it further contains an extract of green or yellow leaves of Gingko biloba, which is known per se. Pharmaceutical composition according to claim 1 or 2, characterized in that the coli extract dosage unit contains 4-9 x 10 ¹² germ-derived metabolites. Pharmaceutical composition according to claim 2 or 3, characterized in that the dosage unit of gingko extract contains about 50-200 mg of extract from dry leaves. 5. Pharmaceutical composition according to any one of claims 1 to 3, characterized in that the ratio of gingcoflavon glycosides to terpene lactones in the dry matter extract is about 5: 1. 6. Pharmaceutical composition according to any one of claims 1 to 3, characterized in that the extracts are in a spatially separated form. 7. Pharmaceutical composition according to any one of claims 1 to 3, further comprising a chemotherapeutic agent for retroviral infections.