DE10021433B4 - Process for the preparation of an antiviral agent - Google Patents

Abstract

method for the preparation of an antiviral agent, characterized that he Tissue containing viruses and antigens, except blood, in Presence of protein cross-linking agents at temperatures from above Heated to 50 ° C.

Description

The The present invention relates to a novel process for the preparation an antiviral agent.

It is known to be viruses not or only insufficient in the human or animal body fought can be. So far, it has not been able to cure HIV-positive patients. This can be attributed to that the Viruses their shape individually and over to change the time are able so that existing Mechanisms of action are turned off.

In the older, Unpublished application of the inventors, a method is described to produce antiviral agents containing viruses and antigens Blood in the presence of protein cross-linking agents as preferably Formaldehyde, p-formaldehyde and / or phenol or phenol derivatives to temperatures of over about 50 ° C heated.

It has now been found that one in viral diseases in which the viruses are insufficient in the Blood, samples of diseased tissues that are infected with viruses are, in a similar manner to the production of antiviral agents can use.

The inventive agent represents an auto-carcinogen z. B. with appropriate workup subcutaneously or perorally the mucous membrane can be administered.

The the patient's own viral-containing tissue is in an amount of a few cubic centimeters, with 1-5 times the amount of physiological Saline or another physiologically acceptable aqueous diluent, by mechanical action, for example ultrasound treatment or high-speed grinders, homogenized and at elevated temperatures in the presence of cross-linking reagents such. B. formaldehyde, p-formaldehyde or phenol. These are genisat in the tissue homo individually cross-linked proteins and thereby denatured, d. H. it is on the virus or the antigen contained in the tissue acted in a specific way, which is surprisingly a means forms, which may be a multiple application to a virus suppression leads.

The Tissue removed during the reaction and then by mechanical action or by chemical anticoagulants such. B. EDTA, heparin and hirudin liquid, to ensure a good distribution of the crosslinking agent.

By The denaturing treatment forms the one anti-pathogen-specific Immune response inducing particles.

to Preparation of a (subcutaneously administered) vaccine is the treated according to the invention Material over Filter with pore sizes of about 400 microns filtered. The liquefied Means may also be given to the patient the mucous membranes of the mouth / throat (gargling).

For the cross-linking of proteins, formalin has proven to be particularly suitable this is homogenized to the tissue in an amount of at least from about 0.1 to about 1.0 volume percent in the form of a saturated formalin added.

The Denaturation temperature is above 50 ° C, preferably between 80 and 85 ° C, maintaining this temperature for about 2 hours.

Of the Effect of the composition of the invention is underlying.

Of the Term autovaccine originally describes a therapeutic Measure, when effective as an agent against a bacterial, chronic infection Taken from the infection site, depending on the case as a pure culture won and afterwards be changed physically and / or chemically again as Cure to be applied.

in the concrete case means autovaccine as a metaphor that for viral Diseases in which the viruses are located in special tissues, these tissues as a homogenate by simultaneous application of heat and Formaldehyde is treated. This treatment of the fabric material and of the antigen (pathogen) to a change the antigenic properties by denaturing the proteins (heat) at simultaneous cross-linking (formalin or formaldehyde or para-formaldehyde or phenol or its derivatives). This denaturation plus cross-linking leads to, that this Antigen (the pathogen) the immune system in a different than the native Way becomes accessible. It follows that too Viruses that may be in the native state remain undetected or an inadequate form of immune response induce (chronic inflammatory course or similar), repelled can be.

Examples for such Diseases are various tumors or carcinomas that have one viral cause, such as certain sarcomas, melanomas, sarcoids, cervical carcinomas etc. or regionally limited viral tissue diseases such as Crohn's disease referred to as small bowel disease.

These considerations are being studied by both animal and part supported by human patients. Mostly it was chronically persistent or relapsing Infections. One can therefore assume that the antigen was basically accessible to the immune system. After application of the antigen in denatured and cross-linked Form came in most cases in less than four weeks to cure the infection. This can only by the described changed of presentation of the antigen by heat and formaldehyde explained become.

example 1

Viral-related inflammatory Disease of the small intestine (Crohn's disease):

Vorliegendes Process for the preparation of an autovaccine was applied to Treatment of chronic inflammatory Bowel disease Crohn's disease. The production of the autovaccine with according to the latest state of knowledge for Crohn's disease at least co-responsible Human Herpes Virus Type 6 (hereinafter HHV6; see. also ref. 1) was carried out by endoscopically obtained 4 Cubic centimeters of the patient's intestinal mucosa from the inflamed areas. For better Passagierbarkeit the intestinal mucosa with the 3 times the amount of physiological saline (alternatively in commercial Cell culture medium z. RPMI1640 without added serum or others for cell culture suitable media) and by mechanical action (ultrasonic treatment) homogenized. Thereafter, it is mixed with 0.3 vol .-% formalin, 2 h at 80 ° C heated, 24 hours at 37 ° C incubated and passed through a sterile sieve (pore size <400 microns). The sample was for sterility checked (negative) and is ready for use. The application of the autovaccine Subcutaneous, starting with the first day of application about 2 ml and the same amount three more times at a distance of five days was applied. The amount of material that is obtained endoscopically is sufficient to perform a vaccination cycle with four applications. The Autovakzine concerned is exclusively patient specific!

Autovakziniert became a 38 year old Patient who is chronically relapsing since the age of 19 Crohn's disease suffered. Primary manifested these thrusts last during a severe recurrence phase in the form of an abscess (sonographically proven beyond doubt) in the left lower abdomen.

through PCR diagnostics became apparent before auto-vaccination positive failure in detecting HHV6 observed. In the course of About 4 weeks after the beginning of autovaccination the treated Patient a significant improvement in general condition with weight gain (6 kg), a macroscopically evident according to the doctor treating Improvement of the condition and a sonographically almost complete remission the initially fist-sized inflammation characterizing Crohn's disease only a few remaining indicated inflammatory mucosal changes. A PCR detection of HHV6 was only weakly positive. A Spontaneous remission can be excluded, according to the doctors. This statement is supported by the local immunological reaction at the injection site on the thigh (probably so-called DTH reaction) as well as the after each of four applications each occurring pain in the Area of the actual abscess in the left lower abdomen, which (initially hypothetical) for one rapid immunological response (eg activation of more specific cytotoxic cells).

Example 2

Caused by papillomaviruses Equine Sarcoids.

An autovaccine according to the invention was also used with great success for the treatment of papillomavirus-induced (Ref.2) equine sarcoid (usually benign cutaneous tumors). Tissue material used for reprocessing in this case is virus-containing tumor material obtained by surgical interventions. Due to the consistency of the material, it is in this case necessary to first dissect tumor material mechanically by means of a scalpel, razor blade or the like and subsequently by using a so-called Ultra-Turrax device at high speeds (in the specific example 20,000 rpm for a few seconds the technical equipment for homogenization may also be different) to achieve a homogeneous suspension. Excised tumor material can be used to produce an autovaccine, which must be regarded strictly as patient-specific. The autovaccine must be adjusted to a density corresponding to a McFarland approx. 5-6 with the aid of sterile pyrogen-free saline solution. The further treatment with formalin and heating is carried out as in Example 1. The vaccine dose is usually 5 ml each which are applied subcutaneously to four appointments in five-day rhythm. With autovaccines produced in this way, we treated a total of more than 100 animals for the prophylaxis of postoperative tumor recurrence. Of 31 of these animals, more detailed data were collected retrospectively from the 11 veterinary surgeons: the average age of these animals at the time of autovaccination was 7.83 +/- 0.76 years. In 21 animals, the autovaccine was used as a first treatment after surgery to prevent possible recurrences, 10 animals were post-emergence post surgical recurrences with autovaccines, which had been produced from the surgically excised recurrent material. In 27 out of 31 animals (87.1%) no further recurrence was observed after auto-vaccination, the mean observation duration being 15.133 months +/- 1.57 months. In the four animals, which had recurrences despite autovaccine, there was no correlation between the occurrence of new recurrences and the duration of observation. In two of these animals, the recurrent sarcoids were significantly smaller than the primary relapsed sarcoids.

Claims (8)

Process for the preparation of an antiviral agent, characterized in that tissue containing viruses and antigens, with the exception of blood, in the presence of protein cross-linking agents, is heated to temperatures above 50 ° C. Method according to claim 1, characterized in that that the crosslinking agents formaldehyde, p-formaldehyde and / or phenol or phenol derivatives. Method according to claim 1, characterized in that that he prior to cross-linking the tissue after addition of pyrogen-free physiological Saline and optionally mechanically comminuted with the addition of anticoagulants and homogenized. Method according to one of claims 1 to 3, characterized that he performs the mechanical crushing by means of an ultrasonic crusher. Method according to one of claims 1 to 4, characterized that the Tissue homogenate 0.3 to 1.0, preferably 0.5 volume percent of a saturated formalin be added. Method according to one of claims 1 to 5, characterized that he the tissue homogenate in the presence of the cross-linking agent at temperatures of 55 to 85 ° C heated and this temperature keeps about 2 hours. Method according to one of claims 1 to 6, characterized that he the heated mixture for producing a vaccine by a small pore Filter with a pore size of ≤ 400 μm happened. Antiviral agent containing the according to claims 1 to 7 prepared antigen and / or virus-containing preparations.