WO2005103231A2

WO2005103231A2 - Dentritic cells charged with toxic substances in order to treat kidney cell carcinomas

Info

Publication number: WO2005103231A2
Application number: PCT/DE2005/000719
Authority: WO
Inventors: Dirk Weickmann
Original assignee: Toximed Gmbh
Priority date: 2004-04-21
Filing date: 2005-04-20
Publication date: 2005-11-03
Also published as: WO2005103231A3; DE112005001476A5; DE102004019323A1

Abstract

The invention relates to a pharmaceutical active substances which are used to treat kidney cell carcinomas by using dendritic cells, which are cultivated in the presence of leukin and charged with toxins obtained from the poison of the loxosceles, scytodidae or drymusa spiders. The invention also relates to a method for the production thereof.

Description

Dendritic cells loaded with toxic substances for the treatment of renal cell carcinomas

Everyone is responsible for the ingestion of food for the preservation of life

Living beings rely on the range of plants and animals that can be reached. But not everything is safe to eat here.

Many plants and animals use to protect their own life and for their own food, on their special organism and its special

Needs coordinated, so-called biogenic, poisons. These biogenic poisons have found their place in the interplay of different types of life in the course of long development periods.

That is why every adult wild animal still recognizes dangerous plants and poisonous animals in its natural environment.

Plants or animals can have a primarily toxic effect through the production of toxins or can only acquire secondary toxicity by ingesting toxic substances from the living or inanimate environment.

The use of these biogenic poisons began in human history as long ago as they were used to kill prey with poisoned weapons.

From the beginning, however, there were certain precautions to use these poisons safely

Basic knowledge of their treatment and effectiveness is required.

The experiments carried out further, the composition of the chemical

Decoding the build-up of biogenic poisons later led to the targeted search for certain active substances as the actual cause of the effects observed.

In particular, after the demand made by Paracelsus (1493 - 1541) to isolate the active substances from medicinal plants, which contributed to the development of latrochemistry, that is, chemistry with regard to its medical field of application, these efforts should have intensified. Above all, the art of distilling fabrics has been put to the service of research and has provided a variety of essential oils and volatiles. But the methods known at that time were insufficient for the isolation of other active substances or even for their chemical breakdown. It was not until the beginning of the 19th century that the development of technical skills in chemistry had advanced far enough to usher in the era of isolating pure active substances from biological material. First, the differences in the solubility of the investigated substances in different solvents were used to separate the active substances from the accompanying substances. The differences in the distribution behavior between two immiscible liquid phases, in the volatility and in the chemical reactivity were observed, for example with precipitation agents.

The development of chromatographic processes in the middle of the 20th century made possible a huge boom in separation technology, the way to identify active substances for combating diseases. Based on the distribution between a mobile and a stationary liquid phase, on the adsorption, the molecular sieve effects, the ion exchange, the affinity (especially of proteins) for certain chemical compounds (e.g. enzyme substrates) and the mobility of charged molecules in the electric field, a large number became new separation techniques developed

Tumors, the most dangerous and feared diseases of our time, are currently being combated in a very radical and less environmentally friendly way. The following can be used as simple, characteristic keywords: steel, radiation and chemotherapy.

On the one hand, this means that, if reasonably attainable, tumors are in principle cut out with the steel of a knife, burned by a wide range of radiation, or destroyed by so-called chemotherapy with aggressive cytostatics that also attack healthy cells. A spatial limitation of the operating area is not possible with normal treatments with a scalpel or with ionizing radiation. Healthy body cells are inevitably destroyed. The undesirable side effects of chemotherapy are generally known, but in contrast, attempts have been made to enable cancer therapy that deserves its name in a more subtle way. For this purpose, the rich treasure of nature was used. For this purpose, among other things, many highly effective substances in therapeutic doses isolated from toxic organisms are used as medicinal substances A pharmaceutical active ingredient is known from DE 199 61 141 A1, in which it has been found that components of the spider venom from spiders of the Sicaridae family can be used for the treatment of tumor diseases. Mainly, a peptide toxin from the poison of this type of spider, a further antagonistic substance obtained from the poison and / or a combination of these components are used medically. This active ingredient can be used for the treatment of tumor diseases and in parallel or in support of tumor operations and residual tumor tissue can be destroyed. In therapy, genetically modified body cells (tumor cells) can be destroyed, since the active ingredient in question recognizes the changed surface structure of such cells and kills them without complications. cannot be used pharmaceutically.

From DE 100 24 383 A1 it is known to use dendritic cells loaded with toxic substances for the treatment of cancer. The toxic substances are selected here from the venom of scolopenders of the genera Scolopendra and Herriscolopendra, snakes of the genera Bitis and Naja, spiders of the genera Loxosceles, Sicarius and Pholcus, as well as scorpions of the genus Parabuthus and a combination of one or more of these toxins.

Dendritic cells are cells of the immune system that are differentiated from stem cells and provided with dendrites, which detect and destroy genetically degenerate cells. The destruction can take place, for example, by pumping immunopeptide toxins into the genetically degenerate cell by means of the dendrites after docking onto them. The dendritic cell takes on the function of a transport cell. The immunopeptide toxin lyses the degenerated cell. With a weakened immune system, the number of dendritic cells present in the body in interaction with the rest of the immune system is no longer sufficient to effectively fight degenerate cells. In order to treat a locally manifest tumor, the patient's own dendritic cells produced in vitro can be injected into the patient or around the tumor. Stem cells from the spinal cord are preferably used for this. The method according to the invention is based on human or animal stem cells, with human spinal cord stem cells being preferred. Own (autologous) stem cells are preferably used here. The spinal cord stem cells are isolated from the spinal cord by methods known per se. Spinal puncture procedures are most frequently used here. The tissue thus obtained can be taken directly into cell culture, but in general the removed spinal cord is treated in such a way that the cells remain viable for a longer period of time. For this purpose, shock freeze treatment has proven its worth. The cells treated in this way are then stored in liquid nitrogen. The removal and storage are of course carried out under the greatest possible sterility. The spinal cord stem cells are placed in conventional cell culture vessels in a medium suitable for growth. Usual media known to the person skilled in the art for cultivating spinal cord stem cells can be used. DMEM-HarrTs F12 - Medium in combination with RPMI has proven particularly successful. To avoid contamination, antibiotics, preferably mixtures of antibiotics, are added. Antibiotics that can be used include penicillin and streptomycin. Amino acids, such as glutamine, and fetal calf serum or substitutes therefor can be considered as further additives. The cells are cultivated in the incubator at a suitable temperature, the body temperature of approximately 37 ° C. being preferred.

However, this known active ingredient does not work in vivo in combination with special tumors such as:

Renal cell carcinoma,

Renal cell carcinoma (also called kidney carcinoma or adenocarcinoma of the kidney) is a disease in which cancer cells are found in certain tissues of the kidney. Kidney cancer is one of the rarer cancers. It occurs more frequently in men than in women.

The kidneys are two organs of the same type, each lying next to the backbone. The kidneys of an adult are about 13 cm long and 8 cm wide and have one typical bean-shaped shape. There are small tubes in each kidney through which the blood is filtered and cleaned of waste materials. These are excreted in the urine. The urine produced in the kidneys is passed through the ureter into the bladder, which collects the urine and keeps it in the body until urination. Renal cell carcinoma is a cancer of the cell layer that lines these tubules of the kidney. If it is part of the kidney in which the urine is collected and transported to the ureter, the so-called kidney pelvis, or if the ureter is directly affected by cancer, it is called urothelial carcinoma (carcinoma of the transitional epithelium) of the renal pelvis and ureter.

It is the object of the active ingredient according to the invention to cause tumor cells to be killed without complications, particularly in the case of renal cell carcinomas.

This object is achieved by the features in claims 1 to 3 and method claim 23.

It is essentially a pharmaceutical active ingredient that is formed from dendritic cells that are loaded with toxins of the animal species mentioned below in connection with interleukin.

The active compounds according to the invention are obtained from the poison of the following animal species: a) Loxosceles spp. (Hermit spiders) b) Scytodiae spp (food spiders) and c) the Drymusa spider species

Spiders of the genus Loxosceles occur in around 50 species in the warmer to the tropical regions of North America, Central America, South America and the Caribbean. About 20 species can be found in Africa as far as the Mediterranean region and southern Europe.

The animals, usually colored in brown tones, reach a body length (without legs) of 8 to 15 millimeters. These spiders are especially active at night and are very shy. Your network is irregular. As a shelter, they look for dark spots under stones, in their cracks, or under leaves.

Spiders of the genus Scytodidae (spiders) have only six eyes. They owe their name to their hunting method. These spiders catch their prey with ejected threads of glue and then bind them with other glue threads attached in a zigzag fashion.

The genus Drymusa is systematically classified together with the genera Loxosceles, Scytodiae and Sicarius among the six-eyed spiders.

The active substance according to the invention can optionally contain a substance antagonistic or synergistic with the respective toxin or peptide toxin and / or penetrating substance from the total poison cocktail of the animal species in question.

The antagonistic or synergistic substance is preferably a phospholipase or a hyaluronidase or a combination of both substances. It is further preferred that the antagonistic or synergistic substance is a mixture of the phospholipases and hyaluronidases and / or toxins present in other species.

The peptide toxin and the substance which is antagonistic and / or synergistically active for this purpose are preferably obtained from the total poison cocktail by a fractionation process, and it is further preferred that the pharmaceutical active substance contains a peptide toxin and a substance which has an antagonistic or synergistic effect and which consists of different fractions come. As a result, the effect of the pharmaceutical active ingredient can advantageously be tailored to the type and / or size of tumor to be treated.

The peptide toxin, and the substance having an antagonistic and / or synergistic effect, can be obtained by fractionation methods known per se for the separation of proteins from the total poison raw mixture. It is preferred that the peptide toxin and the antagonistic or synergistic acting therefor Substance can be obtained by gel chromatography, HPC, affinity chromatography and / or ion exchange chromatography.

It is also preferred that the peptide toxin is present as an active pharmaceutical ingredient in such an amount that the active ingredient has a destructive effect on tumor cells.

Furthermore, the required proportions are chosen so that the active ingredient according to the invention has no or only a slight toxic effect in the patient to be treated. Of course, the amounts of the active pharmaceutical ingredients must also be matched to the type of tumor to be treated and the physical, possibly also psychological, circumstances of the respective patient. The preliminary tests required for such coordination are to be carried out by the person skilled in the art in the context of animal tests and / or ethically justifiable tests on the patient on the basis of his technical knowledge and ability.

Also preferred is a pharmaceutical active ingredient in which the amount of peptide toxin and the substance having an antagonistic or synergistic effect thereon comprises an amount of peptide toxin and antagonistic or synergistic substance which is selected as a function of the tumor to be treated.

It is further preferred that the pharmaceutical active ingredient according to the invention contains conventional carriers and auxiliary substances, such as antibiotics, antifungals, antituberculotics, agents against parasites, cytostatics, amino acids, enzymes which promote wound healing and / or mitotic inhibitors. Preference is given to penicillin / streptomycin, poiymyxin / genetic mycin (5%), mitopodocide, vinca rosea alkaloids, bromelaina or bromelains.

The peptide toxin and the antagonistic or synergistic substance are used in combination with one another in the pharmaceutical active ingredient according to the invention. However, it is also possible to use the individual substances in active pharmaceutical ingredients and to make the special effects of the substances available for therapeutic use. It is also possible to produce the described active ingredients chemically and synthetically or by recombinant means using genetic engineering methods. As is usual with chemical substances, the present invention also includes derivatives and salts of the substances provided according to the invention. For example, the peptide toxin can comprise one or more additions, substitutions and / or deletions of amino acids, it naturally having to be ensured that the medicinal effect according to the invention is retained.

The active ingredient described is obtained by methods customary in chemical process engineering. This includes in particular fractionation processes; however, other methods can also be used, for example immunological methods, in order to get the desired substances out of the total poison cocktail.

A preferred process for the preparation of an active ingredient according to the invention, comprising at least one peptide toxin and / or at least one substance having an antagonistic or synergistic effect, at least one peptide toxin and / or at least one antagonistic or synergistic substance from the poison of animals of the above under a) to c). mentioned species, has the following steps:

Obtaining a raw poison mixture by methods known per se and fractionating the mixture in order to obtain the peptide toxin and the substances having an antagonistic or synergistic effect in fractions which are as separate as possible; optionally combination of different fractions with one another or with poisons originating from other organisms or with antagonistic or synergistic substances in order to obtain a pharmaceutically active substance

The poisons in question contain various peptide toxins and various substances which have an antagonistic or synergistic effect thereon, and other active substances which are also of medical and therapeutic relevance. All of these substances can be found in a certain ratio to be determined by a person skilled in the art can be used therapeutically in a medical active substance.

It is pointed out that the fractionation process shows, by way of example only, a possibility of obtaining the peptide toxins and the substances having an antagonistic or synergistic effect. Further configurations are possible.

In the method according to the invention, it is also preferred that the raw poison mixture is homogenized before the fractionation, and it is further preferred that the fractions are deep-frozen before further processing and more preferably lyophilized.

Surprisingly, combinations of peptide toxins contained in the animal venom and counter-directed (to this end antagonistic or synergistic) enzymes, or peptide toxins in combination with enzymes, in appropriate concentrations and proportions for the treatment of tumor diseases and in parallel or supportive to tumor operations can be used and it can (residual) tumor tissue be destroyed. Here, at least one peptide toxin or at least one substance acting antagonistically or synergistically comes from the poison of animals of the species mentioned.

For example, according to the invention, the destruction of tumor tissue not detected during the operation and the prevention of local tumor metastasis in the organism can be achieved. On the one hand, genetically defective body cells (tumor cells) can be destroyed in the therapy, since the surface protein structure of such cells has been changed and the phospholipases used according to the invention can recognize or selectively bind tumor cells whose surface structure has been changed, and can lyse. On the other hand, tissue in desired, locally delimited areas - here tumor tissue-predestined tissue areas - can be killed without complications. The mode of operation is based on native, mutually influencing modes of action of the peptide toxins and the antagonistic or synergistic substances present in the animal venom as follows:

Phospholipases are also known as hyaluronidases Penetration enzymes described. It is the case that the enzymes mentioned make tissue more permeable to the active substance according to the invention via digestive functions. In addition, they can recognize genetically defective body cells (tumor cells) and these themselves or by infiltration of necrotic or cytotoxic peptides that are coupled to them are killing. In this invention, antagonistic or synergistic substances are understood to mean, for example, phospholipases and hyaluronidases or peptide toxins from animals of the aforementioned genera, although it is not excluded that other antagonistic or synergistic substances are present in the animal venom, which can also be used according to the invention. Human phospholipases, especially type A phospholipases, would also be conceivable in an autologous blood preparation as a therapy for precursor cancer cells (genetically defective but divisible cells). However, the immune status is so poor that the amount of these phospholipases is not equivalent to the amount of genetically defective cells (Bisswanger H. (1994): Enzymkinetik. VCH Weinheim; Zollner H. (1993): Handbook of Enzyme Inhibitors, Part A. 271 272 and 383-388, VCH Weinheim; Zollner H. (1993): Handbook of Enzyme Inhibitors, Part B, VCH Weinheim).

In order to prevent unwanted cell destruction, according to the invention, depending on the type and size of the tumor to be treated, a comparison can be made with respect to absolute and relative amounts of the components of the active ingredient according to the invention in vitro on living human cells (healthy and tumorous) of the tissue type to be treated. Attention to the tendency to spread is of the greatest importance. This can be clarified in preliminary tests in comparison of the tumor tissue strength to the tissue surrounding the tumor.

The mode of action of total poison or individual substances, separated therefrom by column chromatography and characterized by their molecular weight, can take place by testing them in corresponding healthy and tumorous human cell lines.

According to the present invention, the peptide toxins preferably originate from the same organism as those which have an antagonistic or synergistic effect Substances and / or other active substances optionally contained. In this way, the effective interaction or counterplay of these substances developed by nature can be exploited.

The pharmaceutical active substances according to the invention can be produced in such a way that first a crude poison mixture is obtained by methods known per se and fractionation of the crude poison mixture is carried out by fractionation methods, likewise known per se, for the separation of proteins. This serves the purpose of the peptide toxins and to obtain the antagonistic or synergistic substances for this purpose in as separate a form as possible or in separate fractions. Subsequently, different fractions can be combined to produce a pharmaceutical active ingredient, or individual fractions can be combined with peptide toxins originating from other organisms or substances having an antagonistic or synergistic effect. Individual fractions can also be used to produce an active pharmaceutical ingredient. Hyaluronidases from snake venoms, for example from cobra venoms, can preferably be used as antagonistic substances. This can be combined with one or more fractions from substances obtained from animals of the types mentioned under a) to c).

It is also possible according to the invention to combine the fractions with other suitable active ingredients and / or with carriers and auxiliaries customary in pharmacy for the production of pharmaceutical active ingredients. For the production of the pharmaceutical active substances according to the invention, e.g. specific toxic components (necrotic and cytotoxic peptide toxins) as well as natural antagonistic substances (stop substances) of the phospholipase and hyaluronidase type are selected by column chromatographic purification.

It is currently not possible to offer comprehensive preventive therapy due to the diverse types of pathogenesis of tumor diseases. For locally diagnosed tumors (primary or subsequent), surgical removal comes first. A problem that arises here is that Metastasis when tumor tissue that has not been completely removed comes into contact with air when cut through. This problem can be solved by applying pharmaceutical active substances according to the invention to the cut surfaces during surgical tumor removal. Since the operation does not bring the desired results in the vast majority of cases, the use of the active pharmaceutical ingredients according to the invention is to be regarded as a therapy which is gentle on the patient. In so-called permissible medical attempts at healing in patients who had undergone therapy, positive results were shown.

The dendritic cells loaded with the active substances from the animal species mentioned under a) to c) can be used for the therapy of tumor diseases, especially renal cell carcinomas. The dendritic cells can be used as sole therapy, but also as accompanying therapy, without the risk of the disadvantages of conventional therapy.

The dendritic cells are loaded with one and / or more toxic substances from the poison of the animal species mentioned under a) to c). However, it is also possible to use a combination of at least two of these toxic substances from the poisons mentioned.

The poison of the animals mentioned above contains a whole cocktail of compounds, which include not only peptide toxins, but also toxins with an amido group, which, however, have no protein structure and have, for example, a molecular weight of 30 to 55 kDa. The poison cocktail also includes substances with an antagonistic effect, which ensure the spatially and temporally controlled spread of the toxins released by the animal. The toxins are preferably peptide toxins. The toxins generally have necrotic, cytotoxic and / or apoptotic properties. The toxins can be isolated from the poison cocktails of the animals by methods known per se. However, since this is tedious, it has been shown according to the invention that, after fractionation of the poison cocktail, individual fractions containing the toxins can also be used to load the dendritic cells. The toxins are present in these fractions together with other substances. However, since the fractions are selected in such a way that they contain in particular the substances which are toxic to cells, apoptotic and / or necrotic, it is not absolutely necessary to purify the toxins until they are completely pure. The toxins can also be produced synthetically, and modifications of the toxins present in the animal species mentioned and used according to the invention for loading the dendritic cells are also possible, that is to say derivatives which have been obtained by chemical modifications of the toxins. In the case of peptide toxins, derivatives are to be understood to mean, inter alia, toxins in which one or more amino acids have been added, deleted and / or replaced by other amino acids, the toxic properties of which are of course retained in each case. It should be noted that a toxin can combine not only cytotoxic, but also necrotic and / or apoptotic properties.

The dendritic cells loaded with the toxic substances can be used for the targeted therapy of tumors. Since the dendritic cells actively search for and find tumor cells and attack them with cell-destroying substances, the toxic substances are selected so that they have cytotoxic, necrotic and / or apoptotic properties. Such substances are present in the poison cocktails of the animal species mentioned.

It is also possible to combine the dendritic cells loaded in this way with other substances which can be used for tumor therapy.

Interleukin has proven to be particularly suitable here according to the invention.

The cells are cultivated in the presence of pure interleukin. Interleukin 6, interleukin 12 and interleukin 14 are preferred.

It has been shown that these measures form uniform dendritic cells that can be loaded particularly efficiently with active substances.

The choice of the most suitable interleukin differs from case to case and can be determined and optimized by a person skilled in the art through experiments.

The interleukins are substances belonging to the cytokines that act as signal substances of the immune system. As mediators, you are responsible for Induction and course of the T-cell mediated cytotoxic immune response and the B-cell activation (antibody production)

Interleukin 6 (IL 6) is formed by activated T cells, macrophages, fibroblasts and endothelial cells. It is a glycoprotein with an MG of 21000 D. Its main function is its effect as a “colony stimulating factor (CSF). It is also a growth factor for plasma cells, keratinocytes and mesangium cells. It induces the synthesis of acute phase proteins.

Interleukin 12 (IL 12) is a factor that stimulates natural killer cells.

Interleukin 14 (IL 14) is formed by T cells and B cells and causes a further proliferation of already activated B cells.

Claims

Expectations

Claim 1: Pharmaceutical active ingredient for the treatment of tumors, in particular for renal cell carcinomas, containing in a pharmaceutically effective amount: a) dendritic cells, which are loaded with b) toxins and / or optionally synergistic and / or antagonistic substances from the poison cocktail of Spiders of the genus Loxosceles, and c) the dendritic cells being cultivated in the presence of interleukin

Claim 2: Pharmaceutical active ingredient for the treatment of tumors, in particular for renal cell carcinomas, containing in a pharmaceutically effective amount :: a) dendritic cells, which are loaded with b) toxins and / or optionally synergistic and / or antagonistic substances from the poison cocktail from spiders of the genus Scytodiae, and c) wherein the dendritic cells were cultivated in the presence of interleukin.

Claim 3: Pharmaceutical active ingredient for the treatment of tumors, in particular for renal cell carcinomas, containing in a pharmaceutically effective amount: a) dendritic cells, these being loaded with b) toxins and / or optionally substances which act synergistically and / or antagonistically from the poison cocktail of Spiders of the genus Drymusa, and c) the dendritic cells being cultivated in the presence of interleukin

Claim 4: active pharmaceutical ingredient according to one of claims 1 to 3, characterized in that interleukin 6 is used.

Claim 5: active pharmaceutical ingredient according to one of claims 1 to 3, characterized in that interleukin 12 is used.

Claim 6: active pharmaceutical ingredient according to one of claims 1 to 3, characterized in that interleukin 14 is used.

Claim 7: Active pharmaceutical ingredient according to one of the preceding claims, characterized in that the dendritic cells are of human origin. Claim 8: Pharmaceutical active substance according to one of the preceding claims, characterized in that the toxins and the substance having an antagonistic effect thereon or the substance having a synergistic effect were obtained by a fractionation process from the respective total poison cocktail.

Claim 9: Pharmaceutical active ingredient according to one of the preceding claims, characterized in that the toxins obtained by fractionating the poison cocktail are used together with further substances present in the individual fractions without further purification taking place.

Claim 10: active pharmaceutical ingredient according to one of the preceding claims, characterized in that the toxins are used in pure form.

Claim 11: active pharmaceutical ingredient according to one of the preceding claims, characterized in that the toxins are present in recombined form. Claim 12: active pharmaceutical ingredient according to one of the preceding claims, characterized in that the toxins are peptide toxins.

Claim 13: Pharmaceutical active substance according to Claim 12, characterized in that the peptide toxin and the substance having an antagonistic effect thereon, or the substance having a synergistic action and / or the penetrating substance, were obtained from the respective total poison cocktail by a fractionation process

Claim 14: active pharmaceutical ingredient according to claim 12 or 13, characterized in that the peptide toxin and the substance having an antagonistic effect thereon, or the substance having a synergistic effect and / or the penetrating substance by gel chromatography, HPLC, affinity chromatography and / or ion exchange chromatography from the respective total poison - Cocktail was received

Claim 15: Pharmaceutical active substance according to one of claims 12 to 14, characterized in that the peptide toxin and the antagonistic substance, or the synergistic substance and / or the penetrating substance was obtained as follows: By using a column with an inner diameter of 1 , 5 cm and a height of 50 cm, which tapers to 1.5 mm at the bottom and which contains 20 ml of an AcA 34 same chromatographic gel, matrix: 3% acrylamide 4% agarose, fractionation range 20 to 350 kDa, exclusion limit 750 kDa, bead diameter 60-140 µm is filled. The corresponding poison was applied to the gel in a homogeneous state in 0.25M Tris / HCl, pH 6.5 to 7.3, and 1.92M glycine in distilled, deionized water. When the poison solution has passed through the gel and 165 ml of a solution of 0.25 M Tris / HCl, pH 6.5 to 7.3, and 1.92 M glycine in distilled, deionized water have been applied and the first 15 ml of the run discarded and collected in 4 ml fractions, the peptide toxins are in fractions 1, 2, 4, 7, 9 and 10 and the antagonistic substances or the synergistic substances and / or the penetrating substances are in fractions 3.5 , 6,8,11 and 12.

Claim 16: active pharmaceutical ingredient according to one of claims 12 to 15, characterized in that it contains a peptide toxin and a substance having an antagonistic effect thereon, or a substance having a synergistic effect, which originate from different fractions of the respective total poison cocktail.

Claim 17: Pharmaceutical active substance according to one of claims 12 to 16, characterized in that the amount is selected such that a spatial and / or temporally controlled spread in the tissue is ensured.

Claim 18: active pharmaceutical ingredient according to one of claims 12 to 16, characterized in that the amount of peptide toxin, antagonistic substance or synergistic substance and / or penetrating substance is selected depending on the tumor to be treated. Claim 19: Active pharmaceutical ingredient according to one of the preceding claims, characterized in that it contains conventional carriers and auxiliary substances and / or further active substances.

Claim 20: Active pharmaceutical ingredient according to one of the preceding claims, characterized in that it is isotonic solutions, protein solutions, amino acid solutions and / or germicidal solutions, preferably Ringer's solution, 0.9% NaCl solution, human albumin solution, as usual carriers and auxiliary substances and / or glutamine solution, and that it contains antibiotics, antifungals, antituberculotics, agents against parasites, amino acids, enzymes which favor wound treatment, mitotic inhibitors and / or cytostatics as further active ingredients.

Claim 21: active pharmaceutical ingredient according to one of claims 12 to 18, characterized in that a derivative of the peptide toxin and / or the antagonistically active substance or the synergistically active substance and / or the transit substance is contained in the active substance and / or the peptide toxin, or the substance with an antagonistic effect on this, or the substance with a synergistic effect and / or the penetrating substance is produced chemically and synthetically or by recombinant biotechnological methods and its effect is the active substances contained in the total poison cocktail of the respective animals or the antagonistic or synergistic substances and / or penetrating substances and derivatives correspond to this.

Claim 22: Pharmaceutical active substance according to one of the preceding claims, characterized in that the appropriate type of interleukin is determined for the respective application in preliminary tests.

Claim 23: Process for the preparation of a pharmaceutical agent for the treatment of tumors, in particular renal cell carcinomas, with the following steps: a) dendritic cells are cultivated in a pharmaceutically effective amount in the presence of interleukin, b) toxins and / or optionally synergistically with them and / or substances having an antagonistic effect and / or at least one penetrating substance obtained from the poison cocktail of spiders of the genera Loxosceles, Scytodiae or Drymusa, c) the dendritic cells are loaded with these toxins and / or other substances.

Claim 24: Method according to Claim 23, characterized in that the substances with which the dendritic cells are loaded are obtained in the following manner: obtaining a total poison raw mixture by methods known per se and fractionating the mixture to give the peptide toxin or the enzymes and optionally to obtain the antagonistic or synergistic substance and / or the penetrating substance and optionally further substances in fractions separated from one another; Combination of different fractions of the peptide toxin or of the enzymes with fractions which contain antagonistic or synergistic substances and / or penetrating substances for this purpose, or with antagonistic or synergistic substances and / or penetrating substances originating from other organisms, in order to obtain a pharmaceutically active substance receive

Claim 25: Method according to Claim 24, characterized in that the total poison raw mixture is preferably obtained from female animals in each case.

Claim 26: Method according to claim 25, characterized in that the total poison - raw mixture is homogenized before the fractionation.

Claim 27: Method according to Claim 26, characterized in that the fraction is deep-frozen and further preferably lyophilized before further processing to give a pharmaceutically active ingredient.