WO2004075907A9 - Gesundheitsfördernde zusammensetzungen aus lipidhaltigen pilzen und thiocyanaten - Google Patents
Gesundheitsfördernde zusammensetzungen aus lipidhaltigen pilzen und thiocyanatenInfo
- Publication number
- WO2004075907A9 WO2004075907A9 PCT/DE2004/000421 DE2004000421W WO2004075907A9 WO 2004075907 A9 WO2004075907 A9 WO 2004075907A9 DE 2004000421 W DE2004000421 W DE 2004000421W WO 2004075907 A9 WO2004075907 A9 WO 2004075907A9
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- biomass
- thiocyanates
- nanoparticles
- fungi
- compositions according
- Prior art date
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/074—Ganoderma
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
Definitions
- the invention relates to health-promoting agents that consist of biomasses of lipid-containing fungi and thiocyanates and may contain additional active ingredients.
- the biomass can be cultivated in the presence of thiocyanates.
- micro and nanoparticles are obtained, which preferably have an average diameter of 10 nm - 10 ⁇ m. Possible areas of application are medicine, food production, prophylactic use in humans and animals and the therapy of infectious diseases.
- mushrooms there are several options for the use of mushrooms as food or nutritional supplements. The most common is the use of the fruiting body as a food or seasoning. Large-scale production of mushroom mycelium using biotechnological processes is also practiced.
- An example of a biotechnologically obtained product is a health food product made of Fusarium graminearum, which due to its low energy and high fiber values as well as a favorable lipid profile is used directly for human nutrition (Moore-Landecker, E .: Fundamcntals of the fungi, fourth edition, 545, Prentice Hall, Upper Saddle River, New Jersey 07458 (1996); Abbcy, CD.V .: Fungi as source of food; Nutriations and Food Science, 85, 2-9 (1983).
- the lipid content of fungi can vary from less than 1% to 15-20% of the dry weight. On average, around 2 to 8% can be expected.
- Mushroom fat contains all types of lipids such as free fatty acids (mainly unsaturated fatty acids), mono-, di- and triglycerides, sterols (especially ergosterol), sterol esters and phospholipids (Breene, WM: Nutritional and medicinal value of specialty mushrooms, Journal of Food Protection 53: 883-894 (1990).
- Thiocyanates are almost ubiquitous in nature. Thiocyanates are formed in the mammalian organism and are therefore constantly ingested by humans with plant and animal food. In the following situations, an additional intake of thiocyanates was recommended as a health-promoting measure for humans and farm animals (medical and biological significance of thiocyanates (rhodanides) ed. W, Weuffen, VEB Verlag Volk und Pass Berlin 1982): > Vaccination prophylaxis. If the thiocyanate balance is optimized while the antigen is being fed in, the expected immune response should start earlier and be stronger> Increase in the non-specific defense against infections.
- the encapsulation of active substances allows a high active substance loading and a uniform release of the active substance (Eur. J. Pharm and Biopharm 52 (2001) 159-163).
- the encapsulation of the model drug enabled an in vivo release in a period of 8 hours.
- Various manufacturing processes based on lipids are already known for the production of nano- and microparticles. Processes for the production of lipid microparticles based on phospholipids have already been described which have antifungal properties and can be used in the pharmaceutical or cosmetic field (DE 69 00 2905 T2). Other processes describe lipid nanoparticles based on extracted mono-, di- and triglycerides, oils or waxes.
- lipid nanoparticles also describe substances based on lipids for parenteral use (WO 98/56362 AI). Special techniques for the production of lipid nanoparticles are also presented (eg EP 0526 666 A).
- the object of the present invention is therefore to provide new active substances and active substance carriers with improved properties compared to the prior art for various purposes.
- the object was achieved by health-promoting agents consisting of lipid-containing terrestrial mushrooms on the one hand and inorganic thiocyanates or thiocyanates of organic bases on the other.
- the biomasses of the mycelium and / or the fruiting body have been implemented in a cost-effective, direct way with the thiocyanates to give novel substances with special effects.
- the compositions according to the invention can contain further additional active ingredients. It has surprisingly been found that it has been possible to use the health-promoting ingredients of the lipid-containing marine organisms particularly efficiently and to enable various applications which cannot be achieved with the native biomass.
- An advantageous embodiment of the invention consists in using terrestrial fungi cultivated as biomass, the cultivation taking place in the presence of inorganic thiocyanates or thiocyanates of organic bases. During cultivation, it is possible to achieve an enrichment with active ingredients by adding to the culture medium. It is thus possible to provide active substance carriers with improved properties for thiocyanates, vitamins and other active substances for various purposes compared to the prior art, which use the natural ingredients in a special way.
- the biomass of the fungi after conversion into micro and nanoparticles, can serve as an active substance carrier for the minerals and / or free radical scavengers and / or vitamins and / or nutritional supplements and other active substances.
- the invention thus also relates to a composition of biomasses of lipid-containing fungi and mineral substances and / or free radical scavengers and / or food supplements and / or vitamins, in particular vitamin C.
- the conversion to micro- and nanoparticles can advantageously be carried out from the mycelium of the fungi after the biotechnological extraction, the enrichment with active substances being carried out by additions to the culture medium during the biotechnological extraction. It is also possible to enrich with active ingredients during further processing. In particular, it has been possible to develop medicinal feed that does not contain antibiotics.
- fruiting bodies and / or mycelium from one of the fungi listed below: a) Auricularia auricula-judae - Judas ear b) Ganoderma lucidum - shiny lacquer porcelain c) Grifola frondosa - Maitake d) Hericium erinaceus - hedgehog goatee e) Lentinula edodes - Shii-take
- Fungus biomass active ingredient dissolve or suspend Melt aqueous surfactant solution
- the lipid-containing fungi are first heated so that the fatty acids contained therein are liquefied.
- One or more active substances solid or liquid
- the active ingredient is suspended, dispersed or adsorbed in the fatty acids of the lipid-containing fungi.
- a surfactant-water mixture is produced in parallel. This surfactant-water mixture is heated to a temperature above the melting temperature of the fatty acids. The two phases are combined at the selected temperature.
- a pre-suspension is then produced using a stirrer (rotor-stator principle) or using ultrasound.
- the pre-suspension is then homogenized using a high-pressure homogenizer, the number of homogenization cycles and the working pressure being selected according to the desired particle size and stability of the preparation. Between the individual cycles, it must be ensured that the manufacturing temperature is set again and again. The surfactant is used to stabilize the suspension. [0023] If there are problems with the level of the temperature (for example sensitive active substances) during production, it is possible to carry out the entire process even at room temperature. In this case, the process is carried out in the same manner as described above, the active ingredient being adsorbed on the lipid-containing marine microorganisms or being dispersed when a small amount of water is added.
- Fungus biomass active ingredient dissolve or suspend Mixture of active ingredient and fungal biomass in solvent
- the lipid-containing fungi and the active ingredient are suspended in an evaporable organic solvent.
- This mixture is then predispersed (stator-rotor principle or ultrasound), homogenized (high-pressure homogenizer) and then spray-dried or freeze-dried (Scheme 2).
- suitable cryoprotectors are used.
- suitable evaporator e.g. a rotary evaporator.
- the particles from fungi can then be redispersed in suitable aqueous surfactant solutions. After this, redispersion (stator-rotor principle or ultrasound) and homogenization (high-pressure homogenizer) is necessary.
- Co-surfactant org. Solvent dissolve aqueous surfactant solution
- High-pressure homogenizer JJ emulsion solvents evaporate vacuum rotary evaporators drug-laden fungal biomass particles This method is based on the preparation of an emulsion of water and a solution of the biomass from fungus active ingredient in a suitable organic solvent (Scheme 3).
- An emulsifier is used to disperse the active biomass substance.
- the emulsifier and biomass are dissolved in a suitable organic solvent.
- An aqueous phase containing a water-soluble cosurfactant is added to this solution.
- This mixture is then predispersed (stator-rotor principle or ultrasound).
- the organic solvent is removed by evaporation, the biomass containing the active ingredient precipitating in the form of solid particles.
- compositions according to the invention are also effective without the addition of an additional active ingredient, because the valuable ingredients of the mushrooms are made more readily available by the method according to the invention.
- mushroom ingredients such as proteins, minerals and vitamins, polyunsaturated fatty acids, products of secondary metabolism as well as aromatic and flavoring substances in a particularly favorable form.
- the principles presented are also suitable for storing various health-promoting components and / or minerals and / or radical scavengers and / or vitamins and / or nutritional supplements in the biomass.
- ultrafine solid particles in the colloidal size range of approximately 15-1000 nm are produced.
- the biologically active material or the active substance molecule can be enclosed, encased, adsorbed or adhered in the solid matrix (nano-pellet) or shell (nanocapsule) in the dissolved or highly dispersed state.
- one or more active pharmaceutical ingredients can be incorporated into the micro- and nanoparticles.
- compositions of biomasses of lipid-containing fungi in combination with thiocyanates and / or hydrothiocyanates of organic bases can also be used without the conversion into micro and nanoparticles are used and used as health-promoting agents or serve to produce health-promoting agents.
- the preparation according to the invention has the advantage that the release of the incorporated substances can be controlled by choosing the temperature, the active ingredients and the surfactants.
- it is advantageous to disperse the particles in distilled water or in an aqueous medium with additives such as electrolytes, polyons, mono-, di- and polysaccharides, isotonizing agents, buffer substances.
- additives such as electrolytes, polyons, mono-, di- and polysaccharides, isotonizing agents, buffer substances.
- An advantageous embodiment is characterized in that one or more active substances, vitamins or nutritional supplements are added to the biomass in solid and / or liquid form. It is expedient to suspend, disperse or adsorb the added active substances in the fatty acids of the biomass.
- the biomass is combined with a surfactant-water mixture in a further production step. It is advisable to first prepare a pre-suspension with the aid of a stirrer (rotor-stator principle) or with the aid of ultrasound. According to the invention, this pre-suspension is then homogenized with the aid of a high-pressure homogenizer, the number of homogenization cycles and the working pressure being selected according to the particle size and stability of the preparation corresponding to the inventive purpose.
- biomass and active ingredients are suspended in an evaporable organic solvent.
- This mixture is then predispersed (stator-rotor principle or ultrasound) and homogenized (high-pressure homogenizer).
- the solvent is then removed by spray drying or freeze drying or using a rotary evaporator.
- the biomass can be redispersed in suitable aqueous surfactant solutions and then redispersed (stator-rotor principle or ultrasound) and then homogenized (high-pressure homogenizer).
- a coemulsifier or an emulsifier can be used to disperse the biomass / active substance mixture.
- micro and nanoparticles produced by the processes described enable use in a wide variety of fields. Surprisingly, the bioavailability of the new substances was significantly better than that of pure substances.
- the present invention allows for the first time the use of lipid-containing terrestrial mushrooms as active ingredient carriers, for example for antibiotics.
- the invention allows the use of compositions from biomasses of lipid-containing fungi in combination with thiocyanates as health-promoting agents and as food, feed, food supplements and feed supplements. Use in diet products is also possible. A combination with medicinal products is also practicable. It can also be used as a nutritional feed. Further uses according to the invention are the improvement of the rearing results and for the prophylaxis and therapy of infectious diseases in animal husbandry and animal breeding.
- the health-promoting agents can be used in the form of oils, sprays and / ointments and in capsules.
- lipid-containing fungi contain substances which can act as non-specific immunostimulants, ie. H. they stimulate leukocytes and act as activators of the reticoloendothelial system. After the conversion of the biomass of these organisms into micro- or nanoparticles according to the invention, these ingredients enable further applications. For example, use in cover materials for wound care is advantageous.
- Micro- and nanoparticles doped with antibiotics according to the invention allow a controlled release of the antimicrobial active substances and a simultaneous immune stimulation.
- These micro- and nanoparticles can advantageously also be doped with inorganic thiocyanates or hydrothiocyanates with organic bases.
- the invention enables use for the targeted substitution of deficiency states.
- the use of the products produced according to the invention for immune stimulation and for the targeted substitution of the thiocyanate deficiency in dialysis patients is particularly advantageous.
- the particles can be produced by means of stirrers (rotor-stator principle) and high-pressure homogenization, which have been used for decades for the production of fat emulsions for parenteral nutrition and are therefore available for the production of biomass particles on an industrial scale. You are from the State authorities for the production of parenterals are accepted and therefore do not require any new, complex approval procedures.
- the essence of the invention consists of a combination of known (health-promoting and healing effects of certain fungi, health-promoting properties of vitamins and thiocyanates) and new elements (cultivation in thiocyanate-containing nutrient solutions, conversion of the biomass into micro or nanoparticles with the incorporation of Vitamins and other active ingredients, in particular thiocyanates), which mutually influence one another and, in their new overall effect, result in a benefit in use and the desired success, which lies in the fact that active ingredient carriers have been made available which make the incorporated active ingredients particularly bioavailable.
- Example 1 Cultivation of various mushrooms of the Ganoderma genus with and without the addition of thiocyanate
- the cultures of G. pfeifferi, G. resinaceum, G. applanatum or G. adspersum were homogenized as finely as possible and then transferred to a sterile 500 ml Erlmeyer flask and pH 5.4 to 300 ml with malt medium refilled.
- a shake culture batch with 10 flasks 10 ml of homogenate were filled up with 200 ml of malt medium in sterile 500 ml flasks.
- the flasks of the mixture were cultivated at room temperature (20 ° C) under natural light conditions. On days 6, 8, 12, 14, 15, 17, 20, 22 and 30, 3 to 5 pistons were harvested in each case.
- the pH was determined, the mycelium and medium were separated from one another by filtration. The mycelium was transferred to crystallization dishes, lyophilized and then the dry weight was determined.
- the mycelium suspension of 100 ml of filled homogenisate was required, which was introduced into a sterile fermenter and made up to 1000 ml with malt medium. This was operated without air supply on a magnetic stirrer, so that the medium was continuously mixed here as well. The cultivation period was 6-18 days.
- NaSCN NaSCN was added to the malt medium in a concentration of 1 ⁇ 10 -3 mol / l.
- the culture was carried out and the products worked up as in the control.
- a cultivation in a SCN 'containing medium is possible, the growth is influenced positively.
- An SCN ' accumulation in the biomass can be guaranteed in this way.
- Example 2 Extracts obtainable from the mycelium of the species Ganoderma pfeifferi after cultivation in fermenters
- the mycelium was separated from the medium by filtration, transferred to tared crystallization dishes, lyophilized analogously to example 2 and the dry weight determined on a precision balance (Sartorius, Göttingen, G).
- the dried mycelia were filled into extraction thimbles and extracted with dichloromethane (E. Merck, Darmstadt) in a Soxhlet apparatus for 24 hours. After evaporation of dichloromethane residues, the mycelium residue was extracted 3 times for 2 h with 80% ethanol (E. Merck, Darmstadt) with constant shaking at room temperature. The mycelial residue was separated with a Büchner funnel and air dried.
- the mixture was then shaken three times for 2 hours at room temperature with distilled water, and the fungal residue was again separated off by means of a Buchner funnel.
- the residue of the cold-water extract was extracted 3 times with distilled water at 70 ° C. and then filtered.
- a cultivation in a 10 1 fermenter allows the production of mycelium in larger quantities.
- Example 3 Carrying out the experiments according to Example 2, but with the addition of OJ-1%
- Example 4 Extracts obtained by extracting the fruiting body or mycelium with a lipophilic solvent
- dichloromethane is best suited to obtain a high yield of biologically active extracts.
- Example 5 Extracts obtained by extracting the culture medium with ethyl acetate
- the medium of the mushroom culture was concentrated in a vacuum rotary evaporator (Büchi Labortechnik AG, Flawil, Switzerland) to about 50 ml and then extracted by repeated shaking with ethyl acetate. The process was repeated until the ethyl acetate phase had no color.
- Extracts are obtained which make up about 6% of the total mycelium mass. The extracts are separated if the extraction is carried out in stages. Tab. 2 Yields of aqueous extracts and share in the total mass of the mycelium
- Example 7 Production of micro- and nanoparticles from mycelium of Shii-take mushrooms Shii-take (Lentinula edodes) were used
- the biomass is heated to a temperature of 50 ° C.
- an aqueous emulsifier solution is heated to the appropriate temperature (50 ° C).
- both phases are combined at the desired homogenization temperature.
- the mixture is processed using an Ultra Turrax T25 from Janke and Kunkel GmbH & Co KG (Staufen, Germany) in an emulsification process at 8000 revolutions per minute and a duration of 30 seconds.
- the suspension is then homogenized four times with a Micron Lab 40 piston gap high-pressure homogenizer (APV-Gaulin, Lübeck) at a pressure of 500 bar and a temperature of 50 ° C.
- AAV-Gaulin, Lübeck Micron Lab 40 piston gap high-pressure homogenizer
- Example 8 Production of micro- and nanoparticles from mycelium Shii-take (Lentinula edodes) -Vitamin C combinations
- the mycelium of the Shii-take fungus (Lentinula edodes) was used.
- Example 9 Production of Microparticles and Nanoparticles from the Fruit Body of the Mushroom Judas Ear (Auricularia auricula-judae) Fruiting bodies of the Judas ear mushroom were used.
- the biomass is dispersed at a temperature of 25 ° C in an aqueous emulsifier solution.
- the mixture is then processed using an Ultra Turrax T25 from Janke and Kunkel GmbH & Co KG (Staufen, Germany) in an emulsification process at 8000 revolutions per minute and a duration of 30 seconds.
- the suspension is then homogenized four times with a Micron Lab 40 piston gap high-pressure homogenizer (APV-Gaulin, Lübeck) at a pressure of 500 bar and a temperature of 50 ° C.
- API-Gaulin, Lübeck Micron Lab 40 piston gap high-pressure homogenizer
- the biomass is dispersed at a temperature of 25 ° C in an aqueous emulsifier solution.
- the mixture is then processed using an Ultra Turrax T25 from Janke and Kunkel GmbH & Co KG (Staufen, Germany) in an emulsification process at 8000 revolutions per minute and a duration of 30 seconds.
- the suspension will then homogenized four times with a Micron Lab 40 piston gap high-pressure homogenizer (APV-Gaulin, Lübeck) at a pressure of 500 bar and a temperature of 50 ° C.
- API-Gaulin, Lübeck Micron Lab 40 piston gap high-pressure homogenizer
- Example 11 Production of micro- and nanoparticles from mycelium of the Maitake mushroom (Grifola frondosa)
- the mycelium of the mushroom Maitakev (Grifola frondosa) was processed into micro and nanoparticles.
- the biomass is dispersed at a temperature of 25 ° C in an aqueous emulsifier solution.
- the mixture is then processed using an Ultra Turrax T25 from Janke and Kunkel GmbH & Co KG (Staufen, Germany) in an emulsification process at 8000 revolutions per minute and a duration of 30 seconds.
- the suspension is then homogenized four times with a Micron Lab 40 piston gap high-pressure homogenizer (APV-Gaulin, Lübeck) at a pressure of 500 bar and a temperature of 50 ° C.
- API-Gaulin, Lübeck Micron Lab 40 piston gap high-pressure homogenizer
- Example 12 Production of micro- and nanoparticles from fruiting bodies of Mai-take (Grifola frondosa) with vitamin C according to the solvent method
- the biomass is dissolved in n-hexane, the lipids being dissolved from the biomass.
- the biomass is stirred in the solvent for about 3 hours.
- the solvent is then removed using a rotary evaporator.
- a lipid layer forms on the piston surface.
- a solution containing vitamin C is required for redispersion: vitamin C is placed in a Plantacare solution and homogenized four times after a 90-second dispersion process.
- the resulting vitamin C-containing solution is placed in the flask with the lipid layer.
- the lipid layer loosens during the subsequent rotation of the piston and thereby forms encapsulated micro and nanoparticles.
- the rotation takes about 10 hours at a temperature of 40 ° C.
- the biomass is heated to a temperature of 50 ° C. and then the marine active ingredient norhchexanthone used is dispersed or dissolved therein. Separately, an aqueous emulsifier solution is heated to the appropriate temperature (50 ° C). The two phases are then combined at the desired homogenization temperature.
- the mixture is then processed using an Ultra Turrax T25 from Janke and Kunkel GmbH & Co KG (Staufen, Germany) in an emulsification process at 8000 revolutions per minute and a duration of 30 seconds.
- the suspension is then homogenized four times with a Micron Lab 40 piston gap high-pressure homogenizer (APV-Gaulin, Lübeck) at a pressure of 500 bar and a temperature of 50 ° C.
- API-Gaulin, Lübeck Micron Lab 40 piston gap high-pressure homogenizer
- Example 14 Production of micro- and nanoparticles from fruiting bodies of the mushroom Maitake - Vitamin E
- the vitamin E is dispersed in the biomass.
- An aqueous emulsifier solution is prepared separately.
- the mixture is then processed using an Ultra Turrax T25 from Janke and Kunkel GmbH & Co KG (Staufen, Germany) in an emulsification process at 8000 revolutions per minute and a duration of 30 seconds.
- the suspension is then homogenized four times with a Micron Lab 40 piston gap high-pressure homogenizer (APV-Gaulin, Lübeck) at a pressure of 500 bar and a temperature of 50 ° C.
- API-Gaulin, Lübeck Micron Lab 40 piston gap high-pressure homogenizer
- the provitamin Q10 is dispersed in the biomass.
- An aqueous emulsifier solution is prepared separately.
- the mixture is then processed using an Ultra Turrax T25 from Janke and Kunkel GmbH & Co KG (Staufen, Germany) in an emulsification process at 8000 revolutions per minute and a duration of 30 seconds.
- the suspension is then mixed with a Micron piston gap high-pressure homogenizer Lab 40 (APV-Gaulin, Lübeck) homogenized four times at a pressure of 500 bar and a temperature of 50 ° C.
- API-Gaulin, Lübeck Micron piston gap high-pressure homogenizer Lab 40
- Example 16 Testing of micro- and nanoparticles from mycelium of Shii-take mushrooms / vitamin C "produced according to Examples 7 and 8 on an animal model (cow udder teat)
- the udder teats were processed 1 hour after killing the cow.
- the teat was removed from the fat layer.
- the teats were then placed on metal rods of the appropriate size and fastened with clamps.
- the teats were rubbed with 70% alcohol.
- 20 ⁇ l of test substance was pipetted on and triturated with a glass spatula and dried at room temperature for 30-45 minutes.
- the contamination was carried out with 10 ⁇ l of the North German strain, MF 0.5 diluted 1:10. After incubation at 30 ° C for 1.5 hours, the skin areas were spread on Müller-Hinton plates and incubated at 37 ° C.
- m In (number of samples without detection of bacteria / total number of samples).
- Example 17 Testing of micro- and nanoparticles from mycelium of Shii-take mushrooms / vitamin C "produced according to Examples 7 and 8 on the animal model mouse ear
- Mouse ears were used as the test model.
- the donor animals as a source of infection remained untreated.
- Accepted animals were treated with "Shii-take mushrooms / vitamin C" prepared according to Example 8 once a day.
- 10 ⁇ l of test substance was pipetted on and ground with a glass spatula and dried at room temperature
- the donor animals were contaminated with 5 ⁇ l of the North German strain, MF 0.5 diluted 1:10, for which an untreated ear was contaminated and then incubated for 90 minutes at 30 ° C.
- the ears treated with biomass were stamped accordingly The contaminated ears were pressed onto the untreated ears for 10 seconds with pressure.
- the testing of the micro- and nanoparticles, produced according to Examples 7 and 8, also showed a significant reduction in the number of transmitted germs in the donor-acceptor test, with which the transmission of infection was simulated by skin contact.
- the number of repetitions on the mouse ear was 163 for the untreated controls, germ growth was detected on skin areas.
- Example 18 Testing of micro- and nanoparticles "Mycelium of Shii-take mushrooms / vitamin C" produced according to Examples 7 and 8 on the pot-bellied pig Methodology: A pot-bellied pig was available for testing. 2 hours after killing the animal, skin was cut out from the underside of the belly, near the teats, the fat layer was largely removed and cut into pieces. This was stretched over devices so that approx. 1 cm 2 was available for processing. These skin areas were shaved or the bristles cut with scissors and then rubbed twice with 70% ethanol. 20 ul test substance was applied and incubated for 45 min. The contamination was carried out with 10 ⁇ l of the North German strain, MF 0.5 diluted 1:10. After incubation at 30 ° C for 1.5 hours, the skin areas were spread on Müller Hinton plates and incubated at 37 ° C. After the incubation, the germs were counted.
- the number of repetitions on the cow udder teat was 158 for the untreated controls.
- the number of skin areas with germ detection was 2 after application of the micro- and nanoparticles and without germ detection 4. This means that after treatment with the preparation according to the invention according to example 8 on average, only 0.4 germs / cm are to be expected.
- the microbial count reduction is highly significant in the chi-square test.
- Example 20 Testing of micro- and nanoparticles from the fruiting body of Maitake (Grifolia frondosa) / Vitamin C "produced according to Example 12 on the skin model according to Example 17
- Results The testing of the micro- and nanoparticles, produced according to Example 12, showed a significant reduction in the number of transmitted germs in the donor-acceptor test, which was used to simulate the transmission of infection by skin contact.
- the number of skin areas with germ detection was 1 and without germ detection 5. It follows that after the pretreatment according to the invention, on average only 0J8 germs are observed. The reduction of the transmitted germs is highly significant in the chi-square test.
- Example 21 Testing of micro- and nanoparticles, produced according to the examples on the skin model according to example 9
- Example 13 The micro- and nanoparticles Norlichexathon-Vitamin E-Q10 (solvent method) were prepared according to Example 13, 14, 15 and mixed in a ratio of 1: 1: 1. The test was carried out according to the method shown in Example 16.
- the number of preparations tested with norhchexanthone was 22.
- the number of skin areas with germ detection was 19 and without germ detection 3.
- 2 germs are to be expected after pretreatment with preparations according to the invention according to Example 14.
- the biomass (maitake) is heated to a temperature of 50 ° C. and then ubiquinone Q1 is dispersed or dissolved therein. Separately, an aqueous emulsifier solution is heated to the appropriate temperature (50 ° C). Then both phases are combined at the desired homogenization temperature. The mixture is then processed using an Ultra Turrax T25 from Janke and Kunkel GmbH & Co KG (Staufen, Germany) in an emulsification process at 8000 revolutions per minute and a duration of 30 seconds.
- the suspension is then homogenized four times with a Micron Lab 40 piston gap high-pressure homogenizer (APV-Gaulin, Lübeck) at a pressure of 500 bar and a temperature of 50 ° C.
- AMV-Gaulin, Lübeck Micron Lab 40 piston gap high-pressure homogenizer
- Example 23 Prevention of the transmission of MRSA in skin contact with the preparation according to Example 22
- Tails of aseptic mice were used for the experiments. In order to exclude any foreign contamination, it was soaked in 70% alcohol for 5 min before the start of the experiment and then dried under the laminar box.
- the donor mouse tails as a source of infection were contaminated by placing (30 s to 4 min) in a diluted MRSA culture (North German strain, MF standard 0.5 or 0.3) and then incubated at 37 ° C. for 24 h. With smears from these donors, bacterial counts> 100,000 were detected.
- MRSA culture North German strain, MF standard 0.5 or 0.3
- test substances were massaged into the mouse tails, which simulate the receptive organism (acceptors), twice a day.
- the infection was transmitted from the donor to the acceptor by skin contact with the donors for 30 s to 1 min on the shaker at 600 rpm. 2 hours and 24 hours after contamination, the donors were spread on blood Müller-Hinton agar plates. After the agar plates had been incubated at 37 ° C. for 24 h, the colonies were counted.
- Example 24 Particle size determination of the micro and nanoparticles, produced according to Example 8
- Example 25 Detection of the radical scavenging property of the extracts by means of chemiluminescence
- the detection was carried out as follows:
- Example 26 Testing the radical scavenger properties of fungi using the ⁇ -diphenyl- ⁇ -picrylhydrazyl radical scavening effect (DPPH assay)
- the DPPH assay was used to test the radical scavenger properties of the mushroom / vitamin combinations.
- the DPPH radical (2JDiphenyl-l-picrylhydrazyl) has its absorption maximum at 517 nm and a violet color, which is due to the unpaired electron on the nitrogen atom.
- the color changes from violet to yellow when the radical bonds with a hydrogen atom of a radical scavenger and the reduced DPPH-H (2J-diphenyl-ß-picrylhydrazyl) is formed. This also reduces the absorption, which can be measured with a photometer.
- the reference substance is dissolved in ethanol and the following dilutions are made: 10 ⁇ g / ml, 50 ⁇ g / ml, 100 ⁇ g / ml, 500 ⁇ g / ml.
- ImM DPPH dissolved in ethanol corresponds to 294 ⁇ g / ml.
- 4 mg of the mushroom / vitamin combinations are dissolved in 4 ml of ethanol and the following dilution is made: 10 ⁇ g / ml, 50 ⁇ g / ml, 100 ⁇ g / ml, 500 ⁇ g / ml.
- Experimental procedure 500 ⁇ l of the sample are pipetted into an Eppendorf tube.
- Example 28 Use of the extracts to inhibit the activity of proteases
- proteases are involved in numerous post-translational processes of regulating the function of the macroorganism. If the proteases are inhibited, a variety of pharmacological effects can be expected.
- An enzyme inhibition by aqueous and ethanolic extracts according to Examples 2 and 3 was demonstrated using the example of the angiotensin converting enzyme according to Melzig et al., Pharmazie 51 (1996), 501-503. Result: The aqueous and alcoholic extracts have an inhibition of the angiotensin converting enzyme in concentrations of 100-200 ⁇ g / ml.
- Example 29 Use as a vitalizing and germ-reducing additive for pharmaceutical preparations which protects against free radicals
- the addition of the extracts prevents oxidative decomposition, has a preservative effect due to its antibacterial properties and, due to its radical scavenger properties, provides protection against skin-damaging radical formers.
- the Ganoderma extracts have an anti-inflammatory effect.
- the combination of these properties offers good prerequisites for use as an additive for pharmaceutical preparations.
- Example 30 Use of alcoholic and aqueous extracts from the mycelium of G. pfeifferi as vitalizing, pain-relieving and germ-reducing food supplements
- the protection of the cells against toxic radicals can also be of nutritional importance when using nanoparticles obtained from fungi of the genera Auricularia, Ganoderma, Grifola, Hericium and Lentinu la and supplemented with cofactors of antioxidative systems such as thiocyanate and vitamins C and E and trace elements can be important as food supplements.
- This effect which is specific for nanopaticles and is obtained from the biomass of fungi, is supplemented very advantageously by the immunostimulation by thiocyanates, which has been demonstrated by many examples. Immune stimulation is also well documented, particularly in the case of fungi of the genus. So there is a synergistic effect.
- the specific inhibitory effect of the extracts from G. pfeifferi on the neutral endopeptidase can be used for pain relief, since they interfere with the degradation of neuropeptides and endorphins that attack the opiate receptors in the sense of inhibiting the enkephalinase.
- Example 31 Use as a health care product and nutritional supplement
- the neutral endopeptidase and the angiotensin converting enzyme detected in Example G cascade-like functions of the acid organism are interfered with.
- Thiocyanates lead to a higher polarization of the cell membranes.
- the proven inhibition of neutral endopeptidase inhibition according to Melzig et al., Pharmazie 51 (1996), 501-503, causes an increase in renal sodium excretion and thus a reduction in blood pressure.
- Example F The inhibition of the angiotensin converting enzyme demonstrated in Example F also leads, according to Graefe (biochemistry of antibiotics, Spektrum Akademischer Verlag, Heidelberg, Berlin, New York, 1992), to a reduction in blood pressure. Thiocyanate has been used extensively to lower blood pressure. Result: Since in many patients with metabohmic syndrome an increased cholesterol level is combined with an increased blood pressure, the known effects of fungi of the genus Ganoderma in G. pfeifferi are very advantageously supplemented by the hypotensive effect. An inhibition of the serum-mediated lipopylysaccharide binding by 40-60%) was achieved by ethanolic extracts mycelium according to Examples 2 and 3 in concentrations between 0J and 0.2%.
- lipopolysaccharides When infected with Gram-negative bacteria, lipopolysaccharides are released from their cell wall. After binding to the lipopolysaccharide-binding protein, they can stimulate macrophages and mononuclear cells to release endogenous mediators and thus lead to fever, changes in the white blood picture and abnormal widening of the vessels in the periphery of the body.
- Figure 1 Growth curves of the biomass Ganoderma Pfeifferi in the SCN-containing medium
- FIG. 3 growth curves of the biomass Ganoderma applanatum in the SCN-containing medium
- Figure 5 Effect of micro- and nanoparticles from Shii-take mushrooms (produced according to Examples 7 and 8) in comparison to the synergistic combination of Shii-take mushrooms with vitamin C.
- FIG. 6 proof of the interruption of infection chains, simulated in the donor-acceptor model, by the preparation according to the invention according to Examples 7 and 8
- Figure 7 Effect of micro- and nanoparticles from Shii-take produced according to Example 8
- Figure 8 Detection of the interruption of infection chains, simulated in the donor-acceptor model, by the preparation according to the invention according to Example 12
- Figure 9 Particle size distribution of micro and nanoparticles produced according to Example 8.
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Abstract
Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
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EP04715255A EP1603409A2 (de) | 2003-02-27 | 2004-02-27 | Gesundheitsfördernde zusammensetzungen aus lipidhaltigen pilzen und thiocyanaten |
DE112004000789T DE112004000789D2 (de) | 2003-02-27 | 2004-02-27 | Gesundheitsfördernde Zusammensetzungen aus lipidhaltigen Pilzen und Thiocyanaten |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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DE10309681 | 2003-02-27 | ||
DE10309681.7 | 2003-02-27 |
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WO2004075907A2 WO2004075907A2 (de) | 2004-09-10 |
WO2004075907A3 WO2004075907A3 (de) | 2005-01-13 |
WO2004075907A9 true WO2004075907A9 (de) | 2005-02-24 |
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PCT/DE2004/000421 WO2004075907A2 (de) | 2003-02-27 | 2004-02-27 | Gesundheitsfördernde zusammensetzungen aus lipidhaltigen pilzen und thiocyanaten |
Country Status (3)
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EP (1) | EP1603409A2 (de) |
DE (2) | DE112004000789D2 (de) |
WO (1) | WO2004075907A2 (de) |
Cited By (1)
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US9249438B2 (en) | 2001-09-03 | 2016-02-02 | Glycanova As | Production of fungal extracellular immune stimulating compounds |
Families Citing this family (3)
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EP1896600A2 (de) | 2005-06-15 | 2008-03-12 | Medimush A/S | Gegen krebs gerichtete kombinationsbehandlung und kit |
EP1901757A2 (de) * | 2005-06-30 | 2008-03-26 | Ernst-Moritz-Arndt-Universität Greifswald | Präparate auf der basis von naturstoffen |
DE102008035442A1 (de) * | 2008-07-26 | 2010-01-28 | Ernst-Moritz-Arndt-Universität Greifswald | Zubereitungen aus Artemisia Annua, ihre Umwandlung in Mikro- und Nanopartikel und ihre Verwendung |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
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JPS59124984A (ja) * | 1982-12-29 | 1984-07-19 | Morinaga & Co Ltd | 抗酸化性物質及びその製造法 |
DE3502926A1 (de) * | 1985-01-30 | 1986-07-31 | Bayer Ag, 5090 Leverkusen | Thiocyanatomethylthio-gruppen enthaltende verbindungen |
JPS6212721A (ja) * | 1985-07-10 | 1987-01-21 | Yasahiro Morita | 椎茸霊芝の製造法 |
JPS62142119A (ja) * | 1985-12-17 | 1987-06-25 | Hitoshi Nagaoka | アトピ−性皮膚炎治療用クリ−ム組成物 |
WO1988005660A1 (en) * | 1987-01-29 | 1988-08-11 | Co., Ltd. Nikkei | Preparation or mixture of extract of basidiomycetes belonging to genus polyporaceae or lentinus edodes sing. |
DE3942433A1 (de) * | 1989-12-22 | 1991-06-27 | Degussa | Heterocyclische thiocyanate, verfahren zu ihrer herstellung und verwendung als mikrobiozide |
JP2711202B2 (ja) * | 1992-10-06 | 1998-02-10 | 東洋製薬株式会社 | ブドウ球菌抗菌剤 |
JPH08157313A (ja) * | 1994-11-30 | 1996-06-18 | Somar Corp | 工業用防カビ剤 |
DE50002587D1 (de) * | 1999-03-09 | 2003-07-24 | Ganomycin Ges Fuer Biomedizini | Biologisch aktive verbindungen aus ganoderma pfeifferi dms 13239 |
DE19911679C2 (de) * | 1999-03-09 | 2003-06-18 | Ganomycin Ges Fuer Biomedizini | Biologisch aktive Extrakte aus Pilzen der Art Ganoderma pfeifferi |
-
2004
- 2004-02-27 WO PCT/DE2004/000421 patent/WO2004075907A2/de active Application Filing
- 2004-02-27 DE DE112004000789T patent/DE112004000789D2/de not_active Expired - Fee Related
- 2004-02-27 EP EP04715255A patent/EP1603409A2/de not_active Withdrawn
- 2004-02-27 DE DE102004010690A patent/DE102004010690A1/de not_active Ceased
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9249438B2 (en) | 2001-09-03 | 2016-02-02 | Glycanova As | Production of fungal extracellular immune stimulating compounds |
Also Published As
Publication number | Publication date |
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WO2004075907A2 (de) | 2004-09-10 |
EP1603409A2 (de) | 2005-12-14 |
DE112004000789D2 (de) | 2006-07-20 |
DE102004010690A1 (de) | 2004-10-21 |
WO2004075907A3 (de) | 2005-01-13 |
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