Pharmaceutical composition for treating a brain tumor

Abstract

The invention relates to a pharmaceutical composition for treating a brain tumor, containing, in a pharmaceutically active amount, peptide toxin from plants of the genera Dendrocnide and / or Urera and / or Nanocnide and / or Girardinia; and complete toxin or parts of the toxin of plants of the genus Laportea, and to the use of the pharmaceutical composition for treating a brain tumor, to a process for producing the pharmaceutical composition and to a kit of parts comprising the pharmaceutical composition.

Description

[0001] Pharmaceutical composition for the treatment of a brain tumor

[0002] The invention relates to a pharmaceutical composition for treating a brain tumor, comprising a pharmaceutically effective amount of peptide toxin from plants of the genera Dendrocnide and / or Urera and / or Nanocnide and / or Girardinia; and whole toxin or parts of the toxin from plants of the genus Laportea, as well as the use of the pharmaceutical composition for treating a brain tumor, a process for producing the pharmaceutical composition, and a kit of parts containing the pharmaceutical composition.

[0003] - The present invention relates to a pharmaceutical composition, the use of the pharmaceutical composition for treating a brain tumor, a process for preparing the pharmaceutical composition and a kit of parts containing the pharmaceutical composition.

[0004] - In Germany, approximately 9,000 cases of malignant brain tumors occur each year. These include grade II and III astrocytomas, as well as grade IV glioblastomas and glioblastoma multiforme. Even today, brain tumors are usually diagnosed as an incidental finding. However, bleeding in the skull and brain area of ​​patients also regularly occurs, accompanied by loss of consciousness and / or severe, sometimes very severe pain. This makes hospitalization unavoidable.

[0005] - The treatment of brain tumors is extremely difficult. Especially in the case of glioblastomas, the prognosis for patients is usually not positive. Surgical removal of the tumor can extend survival by several months and alleviate symptoms. However, a permanent cure is extremely rare and, from a conventional medical perspective, almost impossible for glioblastomas. - Currently, tumors are considered the most dangerous and feared diseases of our time. They are treated in a very radical and patient-unfriendly manner. Some simple keywords to describe them include:

[0006] Steel, radiation and chemotherapy.

[0007] This means, firstly, that tumors, if reasonably accessible, are surgically removed with a scalpel, destroyed with broad-spectrum radiation, or destroyed through chemotherapy with aggressive chemotherapeutic agents / cytostatics that also attack healthy cells. With both conventional treatments with a scalpel and ionizing radiation, it is not possible to limit the surgical area. Healthy body cells are inevitably destroyed as well. The undesirable side effects of chemotherapy are well known, as are the disadvantages of radiotherapy.

[0008] - Vaccines are not available and, from a cell biological and biochemical point of view, are rather counterproductive when it comes to the use in diseases involving cell degeneration.

[0009] However, the surgical removal of brain tumors in particular has additional side effects. A second operation is often necessary for the patient. The first operation and removal of the brain tumor creates a large scar. In addition to healthy glial cells, diseased, i.e., malignant, glial cells can accumulate there. These are often cells that originate from the tumor and form metastases. This inevitably leads to a diffuse recurrence. This can no longer be treated with conventional methods and is therefore often fatal for the patient.

[0010] - After surgery, radiation therapy and / or chemotherapy usually follow. Cisplatin is often used in various combinations for chemotherapy. Since the beginning of 2002, the oral temozolomide has also been used. IMATINIB is also an oral cytostatic drug that is widely used. In addition to the known side effects such as vomiting, nausea, depression, chemotherapy-induced cancer, and neurological disorders, the problem with the above-mentioned drugs is that they must cross the blood-brain barrier to treat brain tumors. Typically, only 0.4 to 5% of the active ingredients cross the blood-brain barrier. The composition and function of the blood-brain barrier are still only partially understood. This barrier plays a role in protecting the brain from harmful substances, but also enables a regulated energy supply.

[0011] Our body consists of individual organ systems and organs that require different but constant conditions for their function, for example, nutrients, hormones, or electrolytes. All organs are connected by the circulatory system. Since blood contains all the components for the supply and detoxification of the body, filter systems must ensure that only the necessary substances for the individual organ systems are allowed through or partially retained. In this context, we know, among others, the blood-tissue barrier, also known as the blood-parenchymal barrier, the blood-liver barrier, the blood-cerebrospinal fluid barrier, the blood-brain barrier, the cerebrospinal fluid-brain barrier, the blood-nerve barrier, the blood-retinal barrier, and the placental barrier.

[0012] - In these filter mechanisms, a so-called barrier effect prevents or restricts the passage of certain substances from the bloodstream into the respective organ system if these organ systems do not require the components or only require them in lower concentrations. - These "barriers" are not independent organs; rather, they are formed from a multitude of cells and intercellular spaces that allow blood gases, nutrients, and certain chemicals to pass through, or, as endothelial pores, retain macromolecules. They can also act as lipid membranes in the vascular wall, inhibiting the passage of non-lipid-soluble substances or exerting a selective effect on active transport processes in the capillaries. The brain and nervous tissue are protected by two filter systems: the blood-cerebrospinal fluid barrier and the blood-brain barrier.

[0013] The existence and function of the so-called blood-brain barrier has been known for over 100 years and was experimentally demonstrated by Paul Ehrlich as early as 1885. Within the central nervous system, the spaces between neurons are almost entirely filled with glial cells and their processes. The entire metabolism of nerve cells takes place via these glial or endothelial cells. They serve to integrate nerve cells and nerve fibers, as well as to nourish and insulate them. Astrocytes are one type of glial cell. They possess numerous processes with which they attach themselves to the capillary walls and form a virtually seamless endothelial lining that surrounds the capillaries on all sides. These endothelial cells are linked by connecting elements, the "tight junctions," and are equipped with a selective permeability that allows only particles with a diameter of less than 20 nm to pass through.In this way, the entire metabolism of the nerve cells passes through this endothelial network, which, like a biological filter, allows the substances present in the blood to pass through when necessary, but keeps substances harmful to brain function away from the nervous system.

[0014] This endothelial network and the endothelial cells that line the capillaries as a basement membrane are known as the blood-brain barrier. Oxygen, carbon dioxide, D-glucose, D-hexose, some L-amino acids, and lipid-soluble substances necessary for the brain's energy supply pass through unhindered. Degradation products are also released into the blood. The terminal processes of the astrocytes represent a certain barrier to numerous substances, such as certain hormones, non-lipid-soluble, water-soluble, and chemical substances, as well as proteins, thus ensuring the maintenance of a constant environment for the neurons of the nervous system.

[0015] The cell structure of astrocytes is arranged in such a way that it forms an effective barrier against higher-molecular substances and organisms. However, even under normal conditions, it is not completely impermeable, so some particles can always penetrate this barrier. In cases of infection, trauma, inflammation, poisoning, hypoxia, fever, and tumors, the tight junctions between the endothelial cells are expanded by the swelling of the astrocytes and become significantly more permeable to other substances. The change in the width of the junction occurs through the swelling and deflation of the endothelial cells. The basement membrane of the capillaries is also not a closed layer. Depending on the density of the fiber network, pores develop in the membrane, which are actively involved in substance exchange.

[0016] Long before antibiotic treatment became possible, the permeability of the blood-brain barrier was increased by artificially inducing fever, similar to the process that occurs in infections. This was used to treat central nervous system syphilis and to treat shock in psychiatry, allowing drugs to reach the brain directly. After the conditions affecting the blood-brain barrier cease, this temporary permeability returns.

[0017] - Termozolomide, a lipophilic alkylating agent, is currently undergoing clinical trials for the chemotherapy of patients with brain metastases from solid tumors. It crosses the blood-brain barrier and increases the radiosensitivity of tumors during simultaneous radiotherapy. - In contrast, however, attempts have also been made to enable cancer therapy in a more subtle way based on natural substances.

[0018] - For this purpose, many highly effective substances isolated from poisonous organisms are used in therapeutic doses as medicinal substances.

[0019] The use of these biogenic poisons began early in human history. However, safe use of these poisons required a certain basic understanding of their treatment and efficacy from the very beginning. Further attempts to decipher the chemical composition of biogenic poisons later led to the targeted search for specific active ingredients as the actual cause of observed effects.

[0020] - The development of chromatographic techniques in the mid-20th century made a tremendous boom in separation technology, the way to identify active substances for combating diseases, possible. Starting with the distribution between a mobile and a stationary liquid phase, adsorption, molecular sieving effects, ion exchange, and affinity (especially of proteins) to certain chemical compounds (e.g.

[0021] enzyme substrates) and the mobility of charged molecules in the electric field, a variety of new separation techniques have been developed.

[0022] - Especially in recent times, many pharmaceutical active ingredients have been isolated and further developed from biogenic toxins (from fungi, bacteria, plants and animals).

[0023] For example, PCT / EP00 / 12902 discloses a pharmaceutical active ingredient in which it was discovered that components of the venom of spiders of the Sicariidae family can be used to treat tumors. The medicinal use primarily involves a peptide toxin from the venom of this spider species, another antagonistic substance extracted from the venom, and / or a combination of these components. The use of peptide toxins from, for example, the genus Sicarius is proposed for the treatment of breast cancer, lung cancer, adenocarcinomas, liver cancer, and melanomas. However, the treatability of brain tumors such as astrocytomas and glioblastomas is not mentioned. Consequently, the problem of crossing the blood-brain barrier is not discussed either.

[0024] - The active substances described in PCT / EP00 / 12902 can be used for

[0025] It can be used to treat tumor diseases, as well as in parallel or supportive treatment with tumor surgery, and to destroy residual tumor tissue. This therapy can destroy genetically modified body cells (tumor cells) because the active ingredient in question recognizes the altered surface structure of such cells and kills them without complications. The total venom of these spider species, a cocktail of various substances, is not suitable for pharmaceutical use due to its lethal effect even at low doses.

[0026] However, this known drug is not effective in vivo in any combination against a brain tumor, especially not against a specific type of brain tumor, namely oligodendroglioma or oligodendrocytoma. Furthermore, successful therapy requires that this drug largely crosses the blood-brain barrier.

[0027] - Another combination of toxins is disclosed in PCT / EP2006 / 063281. However, the combination of different toxins disclosed therein partially neutralizes each other, so that the desired tumor cell-destroying effect is only inadequately achieved.

[0028] Surprisingly, it has been found that the peptide toxins from Loxosceles degrade the peptide toxins from Latrodectus, preventing the Loxosceles peptide toxins from significantly penetrating the blood-brain barrier. On an electrophoresis gel, after mixing the peptide toxins from Latrodectus and Loxosceles, no distinct bands were observed, but rather a smear caused by fragments of the peptide digestion of the Latrodectus peptide toxins. Spiders of the genus Loxosceles, like spiders of the genus Sicarius, belong to the family Sicariidae. Due to the close relationship between the two spider genera, no effect was expected from a mixture of peptide toxins from the genus Sicarius with those from the genus Latrodectus.

[0029] - It is therefore an object of the present invention to provide a composition which, while overcoming the blood-brain barrier as effectively as possible, brings about a complication-free killing of cancerous body cells from the area of ​​the brain tissue, especially the rare brain tumor oligodendroglioma.

[0030] - Another object of the present invention is to provide a process which enables the preparation of a pharmaceutical composition for the treatment of brain tumors.

[0031] - It is a further object of the present invention to provide a combination of active ingredients which enables effective treatment of brain tumors.

[0032] These objects are achieved by the features of the independent claims. Preferred developments and refinements of the invention can be found in the subclaims.

[0033] - A first aspect of the present invention therefore relates to a pharmaceutical composition for the treatment of a brain tumor, comprising in a pharmaceutically effective amount 1.a) peptide toxin from plants of the genus Dendrocnide and / or plants of the

[0034] Genus IJrera and / or plants of the genus Nanocnide and / or plants of the genus Girardinia and

[0035] 2. b) Peptide toxin from plants of the genus Laportea containing

[0036] The peptide toxin from plants of the genus Dendrocnide and / or plants of the genus Urera and / or plants of the genus Nanocnide and / or plants of the genus Girardinia is capable of destroying tumor cells. Combined with the toxin from plants of the genus Laportea, the tumor-destroying toxin is transported past the blood-brain barrier into the brain, where it can attack the brain tumor. The toxin from plants of the genus Laportea thus acts as a penetrant of the blood-brain barrier. Only this special combination enables the destruction of tumor tissue in the brain.Unexpectedly, the peptide toxins of Laportea were not degraded by the peptide toxins of Dendrocnide and / or Urera and / or Nanocnide and / or Girardinia, so that the Laportea peptide toxins could cross the blood-brain barrier as transfer substances for the other tumor cell-destroying toxins and, in combination with the peptide toxins of Dendrocnide and / or Urera and / or Nanocnide and / or Girardinia, destroy the tumors in the brain area, for example, oligodendrocytes.

[0037] The peptide toxins can be extracted in a conventional manner. For full details, please refer to the description in PCT / EP00 / 12902. Using various chromatographic methods, such as HPLC, the respective total toxin mixtures can be separated into individual fractions, which can then be analyzed for their potency.

[0038] The tumor-destroying effect of individual peptide toxin fractions can be determined in cell culture experiments. The efficacy of the tumor-destroying peptide toxin fractions in combination with the blood-brain barrier penetrating substance from Laportea is then determined in known animal models and cell experiments. Examples of this are given in PCT / EP00 / 12902, which is incorporated herein by reference and whose content is incorporated in its entirety into the present application. As described in PCT / EP00 / 12902 using Sicarius peptide toxins, the skilled person can also isolate the active components for the peptide toxins from the plant genera Dendrocnide, Urera, Nanocnide, and Girardinia and from the total toxin cocktail7 extract of plants of the genus Laportea and test them as described above.In this regard, reference is also made to PCT / EP2006 / 063281, the content of which is also incorporated in its entirety into the present application and to which the applicant refers here. This application shows the use of peptide toxins from Laportea, which are capable of penetrating the blood-brain barrier and simultaneously exhibit the ability to allow other peptide toxins, namely those from Dendrocnide, Urera, Nanocnide, and Girardinia, to penetrate the blood-brain barrier.

[0039] With Laportea, the plant's total toxin / total toxin extract can also be used as an alternative to the peptide toxin fractions. Preferably, only those fractions from the Laportea total toxin cocktail / total toxin extract are used in the pharmaceutical composition that contain the active ingredients capable of passing through the blood-brain barrier or that enable the antitumor agents from Dendrocnide and / or Urera and / or Nanocnide and / or Girardinia to cross this barrier.

[0040] According to a preferred development of the present invention, the pharmaceutical composition additionally contains inactivated Parapoxvirus ovis, preferably of the strain D1701. This is inactivated Parapoxvirus ovis of the strain D1701, to which Polygeline and Aqua ad injectionem have additionally been added.

[0041] According to a further preferred development of the present invention, the plants of the genus Dendrocnide are selected from the group consisting of the species Dendrocnide corallodesme and / or Dendrocnide cordata and / or Dendrocnide cordifolia and / or Dendrocnide excelsa and / or Dendrocnide gigantea and / or Dendrocnide meyeniana and / or Dendrocnide moroides and / or Dendrocnide peltata and / or Dendrocnide sinuata and the plants of the genus Urera are selected from the group consisting of the species Urera caracasana and / or Urera baccifera and / or Urera expansa and / or Urera kaalae llrera nitida and / or Urera simplex and the plants of the genus Nanocnide are selected from the group consisting of the species Nanocnide japonica and / or Nanocnide lobata and / or Nanocnide closii and the plants of the genus Girardinia selected from the group consisting of the species Girardinia bullosa and / or Girardinia diversifolia.

[0042] The plants of the genera Dendrocnide and / or Urera and / or Nanocnide and / or Girardinia and / or Laportea are preferably extracted as follows in order to obtain the total toxic cocktail of the plants:

[0043] The leaves of the plants are harvested at temperatures between 24 and 31 degrees Celsius. The leaves are extracted using common peptide solvents. The resulting total toxin cocktail is separated into its individual components using column chromatography and / or other conventional methods.

[0044] Example 2 describes the extraction of Dendrocnide peptide toxin, Urera peptide toxin, Nanocnide peptide toxin, and Girardinia peptide toxin in more detail. The details for the extraction of peptide toxins from Dendrocnide, Urera, Nanocnide, and Girardinia, which are also applicable to the extraction of peptide toxins from Laportea, are provided in PCT / EP00 / 12902, so that the skilled person can easily obtain the active fractions of the plant (Urticaceae) toxins using this prior art.

[0045] According to another preferred embodiment of the present invention, the peptide toxin from plants of the genus Dendrocnide is the HTDen1 and / or HTDen2 peptide toxin. The preparation of these peptide toxins is described in more detail in Example 2.

[0046] According to another preferred embodiment of the present invention, the composition additionally contains Lachesis D6. Lachesis D6 (Lachesis means pit viper) from the DHU (German Homeopathic Union) is preferably used as a solvent for the dendrocnide peptide toxin and / or the urea peptide toxin and / or the nanocnide peptide toxin and / or the girardinia peptide toxin. Lachesis D6 can be used advantageously, particularly in the preparation of the composition.

[0047] According to another preferred embodiment of the present invention, the plants of the genus Laportea are selected from the group consisting of the species Laportea bulbifera, Laportea canadensis, Laportea cuspidata, Laportea grossa, Laportea interrupts, Laportea mooreana, Laportea ruderalis, Laportea septentrionalis.

[0048] According to a particularly preferred embodiment of the present invention, the plants of the genus Laportea are preferably the species Laportea bulbifera, Laportea canadensis and Laportea ruderalis.

[0049] Mixtures of different toxins from the genera Dendrocnide or Urera or Nanocnide or Girardinia can also be made according to another embodiment of the present invention. The mixture can involve different species of the genus Dendrocnide or Urera or Nanocnide or Girardinia or Laportea, or even mixtures of different populations of a species.

[0050] A second aspect of the present invention relates to the use of a pharmaceutical composition as described above for the manufacture of a medicament for the treatment of brain tumors and glioblastomas, in particular for the treatment of an oligodendroglioma.

[0051] The inventive combination of toxin or parts of the toxin from plants of the genus Dendrocnide or Urera or Nanocnide or Girardinia with a toxin or parts of the toxin from plants of the genus Laportea overcomes the blood-brain barrier. The toxin from the Laportea plants serves as a penetrating substance, allowing the barrier to be penetrated.

[0052] Another aspect of the present invention relates to a method for preparing a composition comprising the steps of: providing an isotonic (0.9%) sodium chloride solution with Dendrocnide peptide toxin and / or Urera peptide toxin and / or Nanocnide peptide toxin and / or Girardinia peptide toxin, optionally adding a further Dendrocnide peptide toxin and / or Urera peptide toxin and / or Nanocnide peptide toxin and / or Girardinia peptide toxin to the sodium chloride solution; and

[0053] - Addition of a total toxin or peptide toxins from plants of the genus Laportea with a molecular weight of 5 kDa to 130 kDa and the ability to penetrate the blood-brain barrier;

[0054] Mix the ingredients, especially by shaking the mixture.

[0055] According to a preferred development of this aspect, an immunomodulator is additionally added before mixing, preferably Parapoxvirus ovis, particularly preferably strain D1701.

[0056] According to another preferred development of this aspect, at least one of the Dendrocnide peptide toxins and / or Urera peptide toxins and / or Nanocnide peptide toxins and / or Girardinia peptide toxins is dissolved in Lachesis D6 before being added to the NaCl solution, preferably both Sicarius peptide toxins.

[0057] According to another preferred development of this aspect, the first dendrocnide peptide toxin is a dendrocnide peptide toxin HTDen1 and / or the second dendrocnide peptide toxin is a dendrocnide peptide toxin HTDen2.

[0058] Furthermore, peptide toxins from Dendrocnide species are preferably used in the pharmaceutical preparation according to the invention as penetrating substances, which facilitate the uptake of the peptide toxins into the tumor cell. Such substances and their extraction are described, for example, in

[0059] PCT / EP00 / 12902. A further aspect of the present invention relates to a kit of parts for the treatment of brain tumors and glioblastomas, comprising a pharmaceutical composition as described above and an extract from frankincense and / or bamboo. The extract from frankincense and / or bamboo leaves is particularly preferred here.

[0060] The main components of frankincense are resins containing boswellic acids and essential oils. In addition to boswellic acids, which belong to the group of pentacyclic triterpenes, the resins also contain a tetracyclic triterpene, tirucalic acid.

[0061] The H15 Ayurmedica preparation is preferably used in the present invention. These are tablets, each containing 400 mg of standardized dry extract of Boswellia serata. The dosage is adjusted as needed. It can vary widely and typically ranges from 3 tablets per week to 25 tablets per day, preferably 10 tablets per week to 20 tablets per day.

[0062] Frankincense extract is used as an alternative to cortisone to prevent cerebral edema during treatment with the pharmaceutical composition according to the present invention. Edema occurs more frequently during the treatment of brain tumors, especially when these tumors are destroyed rapidly, i.e., when they diminish rapidly.

[0063] The different toxins are explained in detail below.

[0064] The tumor cells are destroyed by peptide toxins obtained from the venom of plants of the genera Dendrocnide and / or Urera and / or Nanocnide and / or Girardinia. The preferred species are Dendrocnide corallodesme, Dendrocnide cordata, Dendrocnide cordifolia, Dendrocnide excelsa, Dendrocnide gigantea, Dendrocnide meyeniana, Dendrocnide moroides, Dendrocnide peltata, Dendrocnide sinuata, Urera caracasana, Urera baccifera, Urera expansa, Urera kaalae, Urera nitida, Urera simplex, Nanocnide japonica, Nanocnide lobata, Nanocnide closii, Girardinia bullosa, and Girardinia diversifolia. The peptide toxins can be isolated using known separation methods, such as gel electrophoresis or chromatography, especially column chromatography. The extraction of the crude toxin cocktail / extract from the plants is described below.

[0065] To overcome the blood-brain barrier, active ingredients are preferably extracted from the venom of plants of the genus Laportea. These belong to the nettle family (Urticaceae). They can be found in a wide variety of environments, sometimes even in cities. Many species prefer forests as their habitat.

[0066] The following types are preferred:

[0067] 1.a) Laportea bulbifera

[0068] 2. b) Laportea canadensis

[0069] 3.c) Laportea cuspidata

[0070] 4. d) Laportea grossa

[0071] 5.e) Laportea interrupta

[0072] 6.f) Laportea mooreana

[0073] 7.g) Laportea ruderalis

[0074] 8. h) Laportea septentrionalis

[0075] The two species Laportea bulbifera and / or Laportea ruderalis are particularly preferred for the extraction of peptide substances to overcome the blood-brain barrier when children are to be treated.

[0076] Particularly preferred in the composition according to the present invention are the species Laportea bulbifera and Laportea grossa.

[0077] The Laportea (Laportea canadensis), as used in homeopathy, is found mainly in America along the Pacific coast to Canada.

[0078] Laportea bulbifera occurs in Sri Lanka, India, Bhutan, Myanmar, Thailand, Vietnam, China, Korea, Russia, Japan, and Indonesia. Latrodectus cuspidata occurs in China, Japan, Korea, and Myanmar.

[0079] Laportea grossa is found in RSA.

[0080] Laportea interrupts is found in India, Sri Lanka, Indonesia, Myanmar, Thailand, Malaysia, Vietnam, Yunnan, Taiwan and Australia

[0081] Laportea mooreana

[0082] Laportea ruderalis is known from the Solomon Islands, Christmas Island, Micronesia, French Polynesia, Samoa, Tonga and the Cook Islands

[0083] Laportea septentrionalis

[0084] Laportea venom consists of 5 to 14 different proteins with molecular weights ranging from 15 kDa to 100 kDa. The venom cocktail or parts of the venom cocktail of the Laportea species listed under a) to h) are used, particularly the species Laportea bulbifera and Laportea grossa. The Laportea species listed under a) and g) can also be used on children up to about 12 years of age.

[0085] The substances used in the pharmaceutical composition according to the invention can be obtained naturally from plants. These toxins were originally developed as a defense against predators. This natural mode of action can be maintained by a function-preserving, gentle extraction of the toxin base substance (e.g., by manually harvesting the leaves).

[0086] According to one embodiment of the invention, manual leaf extraction from the plants is provided. This yields genuine, unadulterated native toxins. Analysis and / or quality control of the raw toxin mixture can be performed using electrophoretic methods.

[0087] The room temperature is usually between 25 and 31 degrees Celsius, and the humidity is between 50 and 90 percent. The time of day is irrelevant. It is also possible to produce the described active ingredients contained in the toxins of various plants in recombinant form by chemical synthesis or genetic engineering methods. As is customary with chemical substances, the present invention also encompasses derivatives and salts of the substances provided according to the invention. For example, the peptide toxin can comprise one or more substitutions and / or deletions of amino acids, although it must, of course, be ensured that the medicinal effect according to the invention is retained. The described active ingredient is obtained by methods customary in chemical engineering.

[0088] The active ingredients of the present invention are preferably used in the form of a pharmaceutical composition in which they are mixed with suitable carriers or excipients in doses such that the condition is treated or at least alleviated. Such a composition may contain (in addition to the active ingredients and the carrier) diluents, fillers, salts, buffers, stabilizers, solubilizing agents, and other materials well known in the art. The term "pharmaceutically acceptable" is defined as a nontoxic material that does not interfere with the efficacy of the biological activity of the active ingredient. The choice of carrier depends on the route of administration.

[0089] The pharmaceutical composition may additionally contain other agents that enhance the activity of the active ingredient or complement its activity or use in treatment. Such additional factors and / or agents may be included in the pharmaceutical composition to achieve a synergistic effect or to minimize side effects or undesirable effects.

[0090] Techniques for formulating and administering the compounds of the present application can be found in "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA, latest edition. A therapeutically effective dose further refers to an amount of the compound sufficient to achieve symptomatic improvement, such as treatment, cure, prevention, or amelioration of such conditions. Suitable routes of administration may include, for example, oral, rectal, transmucosal, or intestinal administration and parenteral administration, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, or intranasal injections. Subcutaneous or oral administration to a patient is preferred.

[0091] 1-5 ml, preferably 2-4 ml, of the composition is administered at intervals of approximately 1-4 days. However, the amount of composition administered can be adjusted accordingly by a specialist. This adjustment can also be made during treatment.

[0092] With conventional treatment methods, edema often develops in both primary and secondary or recurrent tumors, causing increased pressure pain. Cortisone is often used to combat edema formation, often in very high doses. However, cortisones can also trigger tumors. Furthermore, cortisone's mode of action can be highly addictive, so discontinuation usually requires a gradual tapering off in multiple, gradually decreasing doses.

[0093] The inventors of the present invention have also discovered that cortisones interfere with the penetration of the compositions of the invention into the brain. Surprisingly, however, the inventors have discovered that frankincense and / or bamboo extract can be used as a complete substitute for edema formation.

[0094] As already mentioned above, H15 preparations are preferred as frankincense extract. Bamboo extract, in particular the extract of bamboo leaves, is also suitable. However, dried frankincense leaves and / or bamboo leaves in capsule form can also be administered directly in combination with the pharmaceutical composition according to the present invention. The following examples illustrate the preparation and mode of action of the pharmaceutical composition according to the present invention. These examples are intended to be exemplary only and do not limit the invention in any way.

[0095] To prepare a pharmaceutical composition in the form of an injection solution or solution for oral administration, first add 1 ml of a saturated solution of dendrocnide peptide toxin HTDen1 in Lachesis D6 DHU (German Homeopathic Union) to 30 ml of a 0.9% NaCl solution. Then, add 1 ml of a saturated solution of dendrocnide peptide toxin HTDen2 in Lachesis D6 DHU. Both steps are preferably carried out at room temperature.

[0096] This mixed cocktail is completed by adding a substance from Laportea bulbifera or Laportea septentrionalis. The toxins from different populations of Laportea spp. can also be used.

[0097] The resulting mixture is then shaken 10 times towards the center of the Earth.

[0098] The resulting composition can be stored at about 7°C under exclusion of light for about 12 months. The experimentally determined

[0099] (electrophoretic) loss of efficacy is about 4% after 12 months.

[0100] In a 1250 ml cell culture flask, the efficacy of the different toxin extracts is determined by counting cells after 24, 48, and 100 hours. It is evident that the cell population decreases only slightly. A decrease in the cell population in vitro corresponds to a reduction in the tumor size in vivo.

[0101] It is evident that after increasing incubation of the cells, hardly any cell growth occurs. Typically, dendrocnide toxin, urera toxin, nanocnide toxin, and Girardinia toxin progressively kill the tumor cells. A peptide toxin derived from dendrocnide is used as the penetrating enzyme. This penetrating substance is available in sufficient quantities and facilitates the diffusion of peptide toxins such as dendrocnide toxin into the tissue and cells.

[0102] Therapy examples

[0103] For the treatment of brain tumors, patients are administered a subcutaneous injection or an oral solution of 2-4 ml of a composition according to the invention every two or three days, depending on the severity of the disease. The course of therapy is monitored using positron emission tomography (PET) and the tumor marker LSA (lipid-bound sialic acid). Therapy can be adjusted accordingly, if necessary.

[0104] In addition, up to 20 tablets of H15 or 3 capsules of frankincense leaves are taken daily to prevent the formation of edema, which can also occur if the tumor is destroyed too quickly.

[0105] The capsules containing frankincense leaves also preferably contain dry extracts of Bambusa ventricosa.

[0106] Treatment process based on various patient examples

[0107] - Patient, born in 1997, diagnosed with glioblastoma at age 5. Inoperable, received only three radiation treatments until July 2004. In August 2004, she began therapy with the composition according to the present invention, initially 2 ml daily for five days, then 2.5 ml every other day until August 2008, and every three days from September 2008. Since September 2004, the girl returned to kindergarten and then to school. The now young woman is working and feeling well. The LSA "started" at 27 and has stabilized at approximately 21.4 since November 2005. The woman receives two frankincense capsules once a week to prevent severe edema. The PET scan has been unremarkable since 2009. The last severe headaches noted were in July 2012.

[0108] Patient, born in 1968, grade III astrocytoma, diagnosed in April 2016.

[0109] In June 2016, he consulted a doctor in Freiburg after his first surgery. He received nine radiation treatments after the surgery and began temozolomide chemotherapy, which he tolerated poorly. By January 2017, a CT scan showed extensive, diffuse tumor spread, with the cell type shifting toward glioblastoma. In May 2017, the patient wanted to begin nettle toxin therapy. He initially began with a daily dose of 5 ml from May 15-22, 2017. Then, until October 15, 2017, he took 4 ml doses orally three times a day (the patient is afraid of needles). From October 17 until now, he has been taking 2.5 ml every two days. The patient's general condition is good, and the PET scan currently no longer shows any tumor. The LSA was approximately 28 after surgery, 25 at the end of October 2017, and has settled at approximately 21 since 2019. The patient has been taking 5 H15 tablets twice daily since May 2017.

[0110] Example 2: General representation of a column chromatographic separation process

[0111] Column chromatographic separation methods are preferred for separating the active peptide toxin fractions from the total toxin cocktails of plants of the genera Dendrocnide, Urera, Nanocnide, Girardinia, and Laportea. A general description of a column chromatographic separation method applicable according to the invention is given below.

[0112] Of course, a person skilled in the art can also use other methods for separating peptide mixtures. The individual fractions obtained by the separation process are tested for their ability to destroy tumor cells, either in cell cultures or in animal experiments. The usability of individual fractions from the Laportea total toxin cocktail as agents that cross the blood-brain barrier is preferably tested in animal models and in modified cell experiments.

[0113] General representation of a column chromatographic separation process:

[0114] To separate the individual components / fractions, the total venom is dissolved in 5 mL of protein solvent for column chromatography, which consists of 0.25 M Tris / HCl, pH 6.5 to 7.3, and 1.92 M glycine in distilled and deionized water. A saturated solution of the total venom cocktail is prepared in a Teflon vial. To ensure homogeneous mixing of the total venom cocktail with the protein solvent, the mixture is vortexed for 60 seconds, avoiding foam formation.

[0115] After homogenization, the solution is poured via a Plexiglas funnel into a transparent, upright Plexiglas column with an inner diameter of 1 cm, a wall thickness of 2 mm, and a height of 50 cm. The column is open but tapered to approximately 1.5 mm at the bottom. The column contains 15 mL of gel (ACA 34; matrix: 3% acrylamide, 4% agarose; fractionation range (MW): proteins from 20-350 kDa, cutoff: 750 kDa, bead diameter: 60-140 μm). The introduced toxin solution displaces the buffer solution present in the gel upon penetrating the gel. After the poison solution has completely penetrated the gel, 165 mL of solvent (0.25 M Tris / HCl, pH 6.5-7.3, 1.92 M glycine) is added to the column in portions (without allowing the gel to run dry). This solvent displaces the poison solution as it passes through the gel. The first 15 mL that drips out of the bottom of the column are the remaining gel buffer and are discarded.Subsequently, 40 fractions of 4 mL each are collected. Due to the chemical and physical properties of this separation system, the individual fractions are 4 mL. SDS-PAGE is used to verify that only one component is present in each fraction. Only one band is detected per fraction.

[0116] For SDS-PAGE, Roti Load 1 + 2 (Carl Roth GmbH & Co KG, Karlsruhe: SDS, glycerol, bromophenol blue, phosphate buffer, Roti Load 1 with mercaptoethanol, Roti Load 2 without mercaptoethanol) is used as loading buffer for the fraction on the gel for peptide binding and protein protection.

[0117] The individual fractions after column separation are collected separately in clean, previously sterilized and screw-capped Teflon vials.

[0118] Example 3: Production of Urticaceae Peptide Toxins. The leaves of the Urticaceae species are extracted as described in more detail above to obtain the total toxin cocktail. The total toxin is then isolated using column chromatography, for example, HPLC. Fractions 2, 7, 8, and 11 contain the brain tumor-killing substances from the Dendrocnide species Dendrocnide moroides and Dendrocnide excelsa. Fractions 5 and 7 contain the brain tumor-killing substances from the Urera species Urera baccifera and Urera nitida.

[0119] Using SDS-PAG electrophoresis and Coomassie blue staining, the following (average) molecular weights of the freeze-dried substances were obtained: Dendrocnide moroides and Dendrocnide excelsa:

[0120] Urera baccifera and Urera nitida:

[0121] Fraction 4 corresponds to the HT1 peptide toxin, fraction 10 to the HT2 peptide toxin.

[0122] The peptide toxins from Girardinia and Nanocnide, like HT1 and HT2, were tested on commercially available standard glioblastoma and astrocytoma cell cultures and also showed the desired effect.

[0123] Kit of parts can be in the form of a kit, a device and / or a system arrangement according to the invention.

Claims

Patent claims 1 . Pharmaceutical composition for the treatment of a brain tumor, comprising in a pharmaceutically effective amount a) peptide toxin from plants of the genus Dendrocnide, plants of the genus Illrera, plants of the genus Nanocnide, plants of the genus Girardinia; and^h) total toxin or peptide toxins from plants of the genus Laportea having a molecular weight of 15 kDa to 100 kDa and the ability to cross the blood-brain barrier.

2. Pharmaceutical composition according to claim 1, characterized in that it additionally contains inactivated Parapoxvirus ovis, preferably strain D1701.

3. Pharmaceutical composition according to one or more of the preceding claims, characterized in that the plants of the genus Dendrocnide are selected from the group consisting of the species Dendrocnide corallodesme and / or Dendrocnide cordata and / or Dendrocnide cordifolia and / or Dendrocnide excelsa and / or Dendrocnide gigantea and / or Dendrocnide meyeniana and / or Dendrocnide moroides and / or Dendrocnide peltata and / or Dendrocnide sinuata and the plants of the genus Urera are selected from the group consisting of the species Urera caracasana and / or Urera baccifera and / or Urera expansa and / or Urera kaalae and / or Urera nitida and / or Urera simplex and the plants of the genus Nanocnide are selected from the group consisting of the species Nanocnide japonica and / or Nanocnide lobata and / or Nanocnide closii and the plants of the genus Girardinia selected from the group,consisting of the species Girardinia bullosa and / or Girardinia diversifolia, with the species Dendrocnide moroides, Dendrocnide excelsa, Urera baccifera, Urera kaalae, Urera nitida and Girardinia diversifolia being particularly preferred.

4. Pharmaceutical composition according to one or more of the preceding claims, characterized in that the peptide oxide from plants of the genus Dendrocnide is the HTDenl and / or HTDen2 peptide toxin.

5. Pharmaceutical composition according to one or more of the preceding claims, characterized in that it additionally contains Lachesis D6.

6. Pharmaceutical composition according to one or more of the preceding claims, characterized in that the plants of the genus Laportea are selected from the group consisting of the species Laportea bulbifera, Laportea canadensis, Laportea cuspidata, Laportea grossa, Laportea interrupta, Laportea mooreana, Laportea ruderalis, Laportea septentrionalis 7. Pharmaceutical composition according to one or more of the preceding claims, characterized in that the plants of the genus Laportea are preferably the species Laportea bulbifera and / or Laportea canadensis and / or Laportea ruderalis.

8. Pharmaceutical composition according to one or more of the preceding claims, characterized in that it further contains peptide toxins from Dendrocnide, Urera, Nanocnide and / or Girardinia.

9. Composition according to any one of claims 1-8 for use in a method of treating brain tumors and glioblastomas.

10. A method for producing a composition according to any one of claims 1-8, comprising the steps of: - providing an isotonic sodium chloride solution with dendrocnide peptide toxin and / or urera peptide toxin and / or nanocnide peptide toxin and / or girardinia peptide toxin; optionally adding another dendrocnide peptide toxin and / or urera peptide toxin, and / or nanocnide peptide toxin and / or girardinia peptide toxin to the sodium chloride solution; and adding a total toxin Z extract or peptide toxins from plants of the genus Laportea, with a molecular weight of 15 kDa to 100 kDa and the ability to penetrate the blood-brain Barrier; mixing of the components, in particular by shaking the mixture.

11. A method according to claim 10, wherein an immunomodulator, preferably inactivated Parapoxvirus ovis, is additionally added prior to shaking.^pj 12. A method according to claim 10 or 11, wherein at least one of the Dendrocnide peptide toxins and / or Urera peptide toxins and / or Nanocnide peptide toxins and / or Girardinia peptide toxins are dissolved in Lachesis D6.

13. The method according to one or more of claims 10-12, wherein the first dendrocnide peptide toxin is a dendrocnide peptide toxin HTDenl.

14. The method according to one or more of claims 10-13, wherein the second dendrocnide peptide toxin is a dendrocnide peptide toxin HTDen2.

15. Kit of parts for the treatment of brain tumors and glioblastomas comprising i) a pharmaceutical composition according to one or more of claims 1-8; and ii) extract of frankincense and / or bamboo

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