Kit of parts for therapy of medulloblastomas and diffuse astrocytomas by means of alcoholic and / or aqueous extracts of various cuscuta spp.

Abstract

The invention relates to a pharmaceutical composition for treatment of a medulloblastoma and / or a diffuse astrocytoma, containing in a pharmaceutically active amount an aqueous or alcoholic extract of plants of the genus Cuscuta, and to the use of the pharmaceutical composition for treatment of a medulloblastoma and / or an astrocytoma, to a method for producing the pharmaceutical composition and to a kit of parts containing the pharmaceutical composition.

Description

[0001] Kit of parts for the treatment of medulloblastomas and diffuse astrocytomas using alcoholic and / or aqueous extracts of various Cuscuta spp.

[0002] The present invention relates to a pharmaceutical composition, the use of the pharmaceutical composition for treating a medulloblastoma and / or a diffuse astrocytoma, a process for preparing the pharmaceutical composition and a kit of parts containing the pharmaceutical composition.

[0003] At least 10,000 cases of malignant brain tumors occur in Germany each year. These include grade II and III astrocytomas, grade IV glioblastomas, glioblastoma multiforme, and medulloblastomas, which primarily occur in children. Even today, brain tumors are still usually diagnosed as an incidental finding. However, bleeding in the skull and brain area of ​​patients also regularly occurs, which is accompanied by loss of consciousness and / or severe, sometimes very severe, pain. Therefore, hospitalization is urgently required.

[0004] Treating brain tumors is extremely difficult. Especially in cases of glioblastoma, the prognosis for patients is generally poor. 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, is currently virtually impossible for glioblastomas.

[0005] Tumors are currently considered the most dangerous and feared diseases of our time. They are treated in a very radical and patient-unfriendly manner. Simple keywords to describe this are: steel, radiation, and chemotherapy. This means that tumors, if reasonably accessible, are essentially surgically excised with a scalpel, destroyed with broad-spectrum radiation, or destroyed via 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 spatially 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.Vaccines are not available, and from a cellular biological and biochemical perspective, their use in diseases involving cellular degeneration is rather counterproductive. 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 predisposes to a diffuse recurrence. This can no longer be treated with conventional methods and is therefore often fatal for the patient.

[0006] 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 these 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.

[0007] 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-parenchyma 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.

[0008] 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.

[0009] 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, forming a nearly seamless endothelial lining that surrounds the capillaries on all sides. These endothelial cells are connected 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.

[0010] 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 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.

[0011] The cell structure of the astrocytes is arranged in such a way that it forms an effective barrier against higher molecular weight substances and organisms. However, even under normal conditions it is not completely impermeable, so that some particles can always penetrate this barrier. In cases of infection, trauma, inflammation, poisoning, hypoxidosis, fever and in the area of ​​tumors, the tight junctions between the endothelial cells are stretched by the swelling of the astrocytes and become significantly more permeable to other substances. The change in the width of the junctions occurs through the swelling and deswelling 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.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 be delivered directly to the brain. After the conditions affecting the blood-brain barrier cease, this temporary permeability returns.

[0012] 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 concurrent radiotherapy.

[0013] In contrast, attempts have also been made to enable cancer therapy in a more subtle way based on natural substances.

[0014] For this purpose, many highly effective substances isolated from poisonous organisms are used in therapeutic doses as medicinal substances.

[0015] 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.

[0016] The development of chromatographic techniques in the middle of the 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 the affinity (especially of proteins) to certain chemical compounds (e.g.

[0017] enzyme substrates) and the mobility of charged molecules in the electric field, a variety of new separation techniques have been developed.

[0018] Especially in recent times, many pharmaceutical active ingredients have been isolated and further developed from biogenic toxins (from fungi, bacteria, plants and animals).

[0019] For example, PCT / EP00 / 12902 discloses a pharmaceutical active ingredient in which components of the venom of spiders of the Sicariidae family have been found to be useful for treating tumors. The medicinal uses primarily involve a peptide toxin from the venom of this spider species, another antagonistic substance derived from the venom, and / or a combination of these components.

[0020] The use of peptide toxins from, for example, the genus Sicarius is proposed for the treatment of breast cancer, lung cancer, adenocarcinoma, liver cancer, and melanoma. 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.

[0021] The active substances described in PCT / EP00 / 12902 can be used to treat tumor diseases, as well as in parallel or adjunctive therapy during tumor surgery, and to destroy residual tumor tissue. Genetically modified body cells (tumor cells) can be destroyed during therapy, as the active substance 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.

[0022] 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 diffuse astrocytoma and / or medulloblastoma. Furthermore, successful therapy requires that this drug largely crosses the blood-brain barrier.

[0023] 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 insufficiently achieved.

[0024] 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.

[0025] It is therefore an object of the present invention to provide a composition which, while overcoming the blood-brain barrier as effectively and as efficiently as possible, brings about a complication-free killing of cancerous body cells from the area of ​​the brain tissue, especially medulloblastomas and / or diffuse astrocytomas.

[0026] Another object of the present invention is to provide a method that enables the preparation of a pharmaceutical composition for the treatment of medulloblastomas and / or diffuse astrocytomas (Mb / d A). It is a further object of the present invention to provide a combination of active ingredients that enables effective treatment of Mb / d A.

[0027] These objects are achieved by the features of the independent claims. Preferred developments and refinements of the invention can be found in the subclaims.

[0028] A first aspect of the present invention therefore relates to a pharmaceutical composition for the treatment of Mb / d A, containing in a pharmaceutically effective amount

[0029] 1. a) aqueous or alcoholic extract of plants of the genus Cuscuta

[0030] An aqueous or alcoholic extract of plants of the genus Cuscuta is capable of destroying cells of an Mb / d A.

[0031] Composition and preparation according to the invention:

[0032] We have now found substances or substance mixtures in various Cuscuta spp., Cuscuta epithymum, Cuscuta epilinum, Cuscuta europaea (Ansbach), Cuscuta campestris, Cuscuta gronovii, and Cuscuta sp. (Italian maquis), that can kill certain brain tumor cells (Mb / d A) in vitro and in vivo. Cuscuta seeds have been used in traditional Chinese medicine to treat various diseases. In our research, we found that a methanol extract or an aqueous extract of the aforementioned Cuscuta species can kill certain brain tumor cells.

[0033] Furthermore, we found that an ethanol extract of the fresh stem areas (Figure 1) of Cuscuta epithymum ssp., incorporated into honey, can kill Mb / d A cells.

[0034] Thus, an ethanolic or aqueous extract incorporated into honey is suitable for the treatment of Mb / d A.

[0035] Approximately 200 species of Cuscuta are known worldwide. Many contain pharmaceutically and medicinally interesting substances. Only a few species have been studied for their therapeutic effects. We investigated extracts of the following Cuscuta species and subspecies for their ability to kill brain tumor cells (especially Mb / d A):

[0036] Cuscuta epilinum

[0037] Cuscuta epithymum ■ Cuscuta europaea (Ansbach)

[0038] ■ Cuscuta europaea (Lyon)

[0039] ■ Cuscuta lupuliformis

[0040] ■ Cuscuta campestris - Cuscuta gronovii

[0041] ■ Cuscuta reflexa

[0042] ■ Cuscuta chinensis

[0043] ■ Cuscuta sp. (Italian Macchie)

[0044] Only extracts of Cuscuta epilinum, Cuscuta epithymum, Cuscuta europaea (Ansbach), Cuscuta campestris, Cuscuta gronovii, and Cuscuta sp. (Italian Macchie) showed a demonstrable effect according to the invention against brain tumor cells (medulloblastoma and / or diffuse astrocytoma cells).

[0045] Particularly preferred in the composition according to the present invention are the

[0046] Species: • Cuscuta epilinum, an endangered species originally distributed between Iran and Central Asia, as well as in Europe. Today, there are still introduced populations in North America.

[0047] • Cuscuta epithymum, widespread worldwide but in sharp decline everywhere

[0048] • Cuscuta europaea, distributed from the temperate zones of Eurasia to northern India

[0049] • Cuscuta europaea (Ansbach), found in the district of Ansbach on the Altmühl

[0050] • Cuscuta campestris, originally widespread in North America, the Caribbean and western South America

[0051] • Cuscuta gronovii, native to North America from Canada to the USA. Also found on Hispaniola.

[0052] • Cuscuta sp. (Italian maquis), found scattered in the maquis between Venice and Istria

[0053] The composition according to the invention is prepared according to one aspect of the present invention in the following manner: a)

[0054] A specific amount of stem is placed in a specific amount of 96% ethanol, or preferably 100% ethanol, or alternatively in a specific amount of injectable water. b)

[0055] The extract from a) is left to stand for at least 24 hours at temperatures of preferably 25 degrees Celsius. The stems are then filtered off. c)

[0056] This active extract from b) is added to 150 mL of liquefied organic blossom honey while stirring continuously. Alternatively, the pure extract can also be used.

[0057] Culture of the Cuscuta sp. preferred according to the invention for optimal active ingredient synthesis:

[0058] To ensure that plants produce optimal quantities of the active ingredients required for the inventive use, it is best to cultivate the respective host plants in plant pots with high-quality plant substrates. More than 30 different components can be detected in the ethanol extract of Cuscuta epithymum, Cuscuta epilinum, Cuscuta europaea (Ansbach), Cuscuta campestris, Cuscuta gronovii, and Cuscuta sp. (Italian maquis). The quantity of active ingredients in the plants can be increased by using the correct substrate for the host plants. Cultivation in plant containers is more efficient than cultivation directly in the soil. When planted directly in the soil, under unfavorable conditions, far too few active ingredients are produced to obtain effective extracts suitable for therapy. For the inventive application, it is important to use the fresh stems of the plants (see drawing 1).

[0059] We observed effects at the following concentrations:

[0060] Therefore, the higher concentrations and the extracts in 100% ethanol are the most effective.

[0061] A further aspect of the present invention relates to a kit of parts for the treatment of brain tumors (Mb / d A), comprising a pharmaceutical composition as described above and an extract from frankincense and / or bamboo. Particular preference is given to using the extract from frankincense and / or bamboo leaves. The main components of frankincense are resins containing boswellic acids and essential oils. In addition to the boswellic acids, which belong to the group of pentacyclic triterpenes, a tetracyclic triterpene, tirucalic acid, is also found in the resins.

[0062] 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.

[0063] 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.

[0064] 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.

[0065] As already mentioned above, H15 preparations are preferred as frankincense extract. Bamboo extract, especially bamboo leaf extract, 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.

[0066] According to a preferred development of this aspect, an immunomodulator is additionally added before mixing, preferably Parapoxvirus ovis, particularly preferably strain D1701.

[0067] Dosage forms:

[0068] It has been shown that the extracts can be taken orally. The active ingredients are absorbed via the digestive tract, and they can also cross the blood-brain barrier. The ethanolic extract dissolves particularly well in honey, so in addition to directly taking the ethanolic and / or aqueous extract neat, it is also advisable to take it dissolved in honey.

[0069] Treatment process based on a patient example:

[0070] Patient, born in 1980, diffuse astrocytoma, diagnosed in May 2016. Came after his 1st

[0071] From surgery to therapy. He received three radiation treatments after the surgery and began temozolomide chemotherapy in parallel. A CT scan showed a strong, diffuse spread of the tumor by March 2018, with the cell type becoming malignant. He initially began with a daily dose of 5 ml of an alcoholic extract with the composition according to the present invention dissolved in honey from June 2 to 30, 2016. Since then, he has been receiving 2.5 ml every two days. The patient's general condition is good, and no new tumor growth is currently visible in the PET scan. The LSA was 28 after the surgery and has stabilized at approximately 19.8 since 2020. The patient has been taking 5 H15 tablets twice daily since summer 2018.

[0072] According to the invention, young (or so-called “fresh”) stems have at least one of the following properties:

[0073] Young stems emerge from the main shoot / stem and are at least 3 to 4 cm long before branching again (see Figure 1). These are usually not yet fully colored and are very light green to yellowish. In the sun, they turn reddish, orange, and yellow after a few days, depending on the host plant and population. The young stems are about 7 days old at harvest.

[0074] Harvesting should ideally take place at temperatures between 20 and 28 degrees Celsius, preferably between 23 and 25 degrees Celsius, if weather permits.

[0075] The weights of Cuscuta stems are listed in the table according to their concentrations. A saturated solution of 10 g of Cuscuta stems per 100 mL of ethanol produces a solution that is best used for incorporation into honey.

[0076] For the purposes of this application, bee honey, organic blossom honey, and flower honey can be used synonymously. Blossom honey refers to honey that is not forest honey and is not collected from just one (main) plant. Thus, it is not (pure) acacia or dandelion honey, but rather from hectares of many plant species.

[0077] The composition and extract according to the invention may comprise an extract of frankincense and / or bamboo.

[0078] Young stem sections 1, 2, 3, 4 are shown in Figure 1; these are preferably to be harvested and used according to the invention.

Claims

Patent claims 1. Extract for combating medulloblastomas and / or diffuse astrocytomas containing stems of Cuscuta epithymum, Cuscuta epilinum, Cuscuta europaea, Cuscuta campestris, Cuscuta gronovii and Cuscuta sp.

2. Extract according to claim 1, characterized in that it contains honey.

3. Extract according to claim 2, characterized in that the honey is in the form of organic honey, bee honey, blossom honey and / or organic flower honey.

4. Extract according to at least one of the preceding claims, characterized in that the stems are young.

5. Extract according to at least one of the preceding claims, characterized in that the extract contains at least 96% ethanol and / or aqua ad injectabilia.

6. Extract according to at least one of the preceding claims, characterized in that it is an ethanolic or aqueous extract.

7. Extract according to at least one of the preceding claims, characterized in that the extract comprises inactivated Parapoxvirus ovis and / or Parapoxvirus ovis strain D1701.

8. A process for preparing a therapeutic composition according to any one of claims 1 to 7.

9. A process for the preparation of a therapeutic composition according to any one of claims 1 to 7, comprising: a) introducing a certain amount of stems of Cuscuta epithymum, Cuscuta epilinum, Cuscuta europaea, Cuscuta campestris, Cuscuta gronovii and Cuscuta sp. into a certain amount of at least 96% ethanol and / or injectable water; b) allowing the extract from a) to stand for at least 15 hours; c) filtering off the leaves; and d) incorporating an active extract obtained from process step c) into a predetermined amount of liquefied honey.

10. A method according to claim 8, characterized in that the resting takes place at temperatures of approximately 23 to 26 degrees Celsius.

11. A method according to claim 8 or 9, characterized in that the resting period lasts between 22 and 30 hours.

12. Method according to one of claims 8 to 10, characterized in that the predetermined amount of liquefied honey is selected such that an absorption saturation occurs when the active extract is incorporated into the honey.

13. A kit of parts for the treatment of brain tumors such as medulloblastomas and / or diffuse astrocytomas, 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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