EP1603409A2

EP1603409A2 - Salutary compositions consisting of fungi containing lipids and thiocyanates

Info

Publication number: EP1603409A2
Application number: EP04715255A
Authority: EP
Inventors: Wolf-Dieter Juelich; Gerold Lukowski; Ulrike Lindequist
Original assignee: Ernst Moritz Arndt Universitaet Greifswald
Current assignee: Universitaet Greifswald
Priority date: 2003-02-27
Filing date: 2004-02-27
Publication date: 2005-12-14
Also published as: WO2004075907A2; DE112004000789D2; DE102004010690A1; WO2004075907A9; WO2004075907A3

Abstract

The invention relates to salutary compositions, which consist of biomasses of fungi containing lipids and thiocyanates and which can contain additional active substances. The biomasses can be cultivated in the presence of thiocyanates. Microparticles and nanoparticles, preferably with an average diameter of between 10 nm and 10 µm can be obtained by the conversion of the biomasses. Potential areas of application are medicine, foodstuff manufacture, a prophylactic application for humans and animals and the treatment of infectious diseases.

Description

Health promoting compositions of Iipid mushrooms and thiocyanates

description

The invention relates to health-promoting compositions which consist of biomasses of lipid-containing fungi and thiocyanates and may contain additional active ingredients. The biomass of the mushrooms can be cultivated in the presence of thiocyanates. By converting the biomass, micro and nanoparticles are obtained, which preferably have an average diameter of 10 nm - 10 μm. Possible areas of application are medicine, food production, prophylactic use in humans and animals and the therapy of infectious diseases.

[0002] 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 .: Fundamentals of the fungi, fourth edition, 545, Prentice Hall, Upper Saddle River, New Jersey 07458 (1996); Abbey, 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 (Chang, ST; Miles, PG: The nutritional attributes and medicinal value of edible mushrooms. In Edible mushrooms and their cultivation, 27-40, CRC Press, BocaRaton (1989); Garcha, HS, Klianna, PK, Soni, GL: Nutritional importance of mushrooms. In: Mushroom Biology and Mushroom Products, ed. Chang, ST, Buswell, JA, Chiu, SW: The Chinese University Press, JonKong, 227-236 (1993); Breene, WM: Nutritional and medicinal value of specialty mushrooms, Journal of Food Protection 53, 883-894 ( 1990); Bötticher, W .: Technologie der Pilzverwertung, Verlag Eugen Ulmer, Stuttgart 21-55, (1974).

[0004] Thiocyanates are almost ubiquitous in nature. Thiocyanates are Mammal organism formed and 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 Gesundheit Berlin 1982):

> Vaccination prophylaxis. If the thiocyanate balance is optimized while the antigen is being fed, the expected immune response should start earlier and be stronger. Increase in the non-specific defense against infections. In phases of increased stress (e.g. premature babies, increased risk of infection in the event of frequent respiratory infections or in the case of virus flu, in the keeping period of animals), the optimization of the immune response with thiocyanates should be promising. Use in diseases with involvement of the immune system. With such diseases, e.g. of rheumatic forms, asthmatic diseases, allergic skin diseases and possibly neoplasms may have therapeutic consequences.

> Protective effect with toxic loads.

Numerous documents have been provided for the fundamental importance of an adequate supply of ttaocyanate with food or feed (Bredereck, G., W.-D. Jülich, W. Weuffen and W. Schindler: Use of inorganic rhodanides in piglet rearing. Arch.exp.Vet.med. 1977; 31: 665 - 670) Rotermund, L., W. Weuffen, W. Kurzweg, A. Kramer and W.-D. Jülich: The protective system in industrial animal production. Mh. Vet. Med. 1978; 33: 524-531

Weuffen, W., W.-D. Jülich, B. Thürkow, H. Blohm, E. Uecker, I. Böning and E. Schüemann: Studies on the influence of thiocyanate (rhodanide) on the serum protein of cattle and pigs, Mh. Vet. Med. 1982; 37: 887 - 889 Jülich, W.-D., and W. Weuffen: Relevance and segregation of the thiocyanate serum level determination in tumor patients depending on the prevalence. Wiss. Z. Ernst-Moritz-.Arndt-Univ. Greifswald, Med. R. 1984; 32: 37-39

Kramer, W., W. Weuffen, H. Schroeder, W.-D. Jülich, U. Grimm, S. Hecht, K. Schirdewan, W. Schwenke, V. Bauernhorst and E. Stark: Thiocyanate levels in serum and erythrocytes in patients with endogenous or contact eczema and cutan-vascular allergy. Wiss. Z.

Ernst-Moritz-Arndt-Univ. Greifswald, Med. R. 1984; 33: 40-42

Hübner, G., and W.-D. Jülich: On the influence of thiocyanate on the thyroid gland. Z.

Ernst-Moritz-Arndt-Univ. Greifswald, Med. R. 1984; 32: 26-27 Kramer, A., W. Weuffen, A. Apitzsch and W.-D. Jülich: Stimulation of Antibodies and Liver

Protection with Thiocyanate during Rabies Vaccination Annais of Allergy. , 1985; 55: 405

Weuffen, W., A. Kramer, H. Ambrosius, V. Adrian, H. Below, W.-D. Jülich, St. Koch, B.

Thürkow and F. Verbeck: On the importance of the endogenous active ingredient and environmental factor thiocyanate. Zbl. for hygiene and environmental medicine 1990; 423: 615-652 Weuffen, W., A. Kramer, H. Below, H. Böhland, W.-D. Jülich, B. Thürkow and U. Burth:

Thiocyanate ion as a physiologically important active ingredient in living nature. pharmacy

1990; 45: 16-30

Kramer, A., W.-D. Jülich and A. Schulz: Alimentary significance of Thiocyanat.Hyg. Med.

1993; 18: 56 Jülich, W.-D., A. Kramer, V. Hingst, H.-G. Sonntag, A. Schulz, M. Daut, G. Bahlmann: Zum

Influence of a thiocyanate-rich diet on hepatitis B vaccination, especially in problem patients in hemodialysis. Hyg. Med. 1996; 21: 34-35

Kramer, A., M. Völzke, N. Völzke, B. Thürkow, T. Hiepe, W.-D. Jülich, A. Weber and W.

Weuffen: Promotion of wool formation and body mass development in sheep through aliminary thiocyanate supplementation. Berliner Tierärztl. Wschr., 1996; 109: 419-428

Kramer, A., M. Kühn, W.-D. Jülich: Reduction of those induced by cyclophosphamide

Leucopoies suppression by thiocyanate. Hyg. Med. 1996; 21:34

Jülich, W.-D., A. Kramer, V. Hingst, H.-G. Sunday, C, Schürt, M. Daut, A. Schulz, G.

Bahlmann, H. Zöllner, R. Hampel, E. Panzig: Investigations on the stimulation of the immune response in the case of hepatitis B vaccination by an alimentary in thiocyanate supplementation in healthy subjects and in dialysis patients. Umweltmed Research Practice

1997; 2: 71-83

The use of the thiocyanates for the objectives recommended by Weuffen has not so far been successful despite the well-documented positive effects. A major reason should therefore be the difficulty in finding a suitable form of application for the thiocyanates. Attempts have therefore even been made to achieve an enrichment of thiocyanate and to use corresponding products for dietetic purposes (Zöllner, H., W.-D. Jülich, Ch. Bimek, A. Kramer: Influence of thiocyanate bound to polycationic carriers on the yield of spelled and the thiocyanate content in the grain. Deutsche Lebensmittel-Rundschau 96 (2000), 103-107; Zöllner, H., A. Kramer, W.-D. Jülich, Ch. Bimek: Increasing the yield and improving the quality of speit after fertilization with thiocyanate. J. Plant Nutr. Soil Be. 164: 105-106 (2001).

The encapsulation of active ingredients allows a high active ingredient loading and a uniform release of the active ingredient (Eva. 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. Active pharmaceutical ingredients are also encapsulated with the help of these lipids obtained (WO 94/20072 AI). Other 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 0 526 666 A).

Disadvantages of the prior art

In particular, the use of antibiotics, which are also used in human medicine, as medicinal feed has rightly come under increasing criticism. [0009] No feed supplement used hitherto meets all the requirements of animal breeding with regard to effectiveness and avoidance of side effects.

Even in the field of human nutrition, it has so far not been possible to use the vitalizing effects of physiological active ingredients such as thiocyanate and natural ingredients to a sufficient extent.

The object of the present invention is therefore to provide new active ingredients and active ingredient carriers with improved properties over the prior art for various purposes.

[0012] The object was achieved by health-promoting compositions consisting of lipid-containing terrestrial fungi on the one hand and inorganic thiocyanates or thiocyanates of organic bases on the other. According to the invention, 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. [0015] An advantageous embodiment of the invention consists in using terrestrial mushrooms 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.

In this way it has been possible to use the health-promoting ingredients of the mushrooms particularly efficiently. At the same time, 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. This enables various applications that cannot be achieved with the individual components. The invention thus also relates to a composition of biomasses of lipid-containing fungi and minerals and / or radical scavengers and / or nutritional supplements and / or vitamins, in particular vitamin C.

The conversion into 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 bioteclinological 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.

For various applications, it is advantageous to use 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

The conversion into micro and nanoparticles leads to new, valuable products that cannot be achieved in known ways and contains three alternatives:

1. Homogenization process

Scheme 1: Manufacturing process of drug-laden fungal biomass particles (homogenization principle)

Fungus biomass active ingredient

Loosen or suspend

Melt aqueous surfactant solution

Stirring (rotor stator) or ultrasound

dispersion

High-pressure homogenizer 1 1 drug-laden fungal biomass particles

The lipid-containing fungi are first heated so that the fatty acids contained therein are liquefied. One or more active substances (solid or liquid) are added to this biomass (Scheme 1). 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. Then use a stirrer

(Rotor-stator principle) or with the help of ultrasound a pre-suspension is produced. The pre-suspension is then homogenized with the aid of a high-pressure liomogenizer, 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. 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.

Scheme 2: Manufacturing process of biomass particles (solvent homogenization process)

Fungus biomass active ingredient

Loosen or suspend

Mixture of active ingredient and fungal biomass in solvent

predispersal

high-pressure homogenizer

Redispersion of the fungal biomass particles in aqueous surfactant solution

Stir or ultrasound

Nachdispergierungsverfahren

high-pressure homogenizer

&

Enddispergierungsverfahren

Active ingredient biomass particles 2. Solvent homogenization process

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). When freeze drying, it should be noted that suitable cryoprotectors are used. It is also possible to remove the organic solvent using a 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.

3. Solvent emulsion process

Scheme 3: Manufacturing process using the solvent emulsion process

Mushroom biomass + active ingredient emulsifier

Co-surfactant org. solvent

JJ ^lots

Solution aqueous surfactant solution

Rotor stator or ultrasound

emulsion

high-pressure homogenizer

&

emulsion

Solvents evaporate vacuum rotary evaporators containing mushroom biomass particles loaded with active substances 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). After a homogenization step using a high-pressure homogenizer, the organic solvent is removed by evaporation, the biomass containing the active ingredient precipitating in the form of solid particles.

The use of these methods leads to novel biomass active ingredient - particles with an average diameter that is between 10 nm and 10 microns depending on the type of manufacture.

The 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. In this way it is possible to use 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. With the method according to the invention, 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.

[0029] In addition, according to the invention, one or more active pharmaceutical ingredients can be incorporated into the micro- and nanoparticles.

According to the invention, the 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. According to the invention, 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. To achieve the purpose according to the invention, it may be necessary to add one or more substances that stabilize the dispersion. 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. According to the invention, 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.

In another manufacturing method according to the invention, 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. If necessary, the biomass can be redispersed in suitable aqueous surfactant solutions and then redispersed (stator-rotor principle or ultrasound) and then homogenized (high-pressure homogenizer). If necessary, a coemulsifier or an emulsifier can be used to disperse the biomass / active substance mixture.

The micro and nanoparticles produced by the processes described enable use in a wide variety of fields. Surprisingly, the bioavailability of the new substances compared to the pure substances was significantly better. 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.

[0038] Numerous 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.

It is particularly advantageous that the particles can be produced by means of agitators (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 features of the invention emerge from the elements of the claims and from the description, both individual features and several in the form of combinations representing advantageous designs for which protection is sought with this document.

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 biomasses into micro or nanoparticles with incoφeration 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 were able to be made available which make the incorporated active ingredients particularly bioavailable.

The invention is to be explained on the basis of exemplary embodiments, without being limited to these examples.

embodiments

Example 1: Cultivation of various mushrooms of the Ganoderma genus with and without the addition of thiocyanate

The growth of G. pfeifferi, G. resinaceum, G. applanatum and G. adspersum with and without the addition of thiocyanate was examined comparably. The same culture conditions were observed for all mushrooms. [0045] All cultivation steps were carried out under aseptic conditions. The preculture was carried out on solid and later in liquid Hagem medium. For this purpose, small pieces were transferred from a slant agar tube to the Hagem agar by means of sterile vaccination and spread out. After a growth time of approx. 3 weeks with a natural day-night rhythm, pieces were again punched out of this agar, transferred to sterile 100 ml Erlmeyer flasks and incubated with 50 ml liquid Hagem medium pH 5.4. The stand culture could be used for the following experiments after 2-3 weeks. Shake culture and fermenter culture

With a homogenizer, the cultures of G. pfeifferi, G. resinaceum, G. applanatum and 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. For 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. After 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.

In order to produce a fermenter culture, 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.

Cultivation with the addition of thiocyanate

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.

Result [0050] A cultivation in a SCN ^'containing medium is possible, the growth is infiuenced positive. An SCN ^" enrichment 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

Methodology: Freshly harvested young fruiting bodies of the species G. pfeifferi were cleaned and broken up under sterile conditions. Tissue pieces were removed from the area of the transition from the hat to the stem using sterile tweezers and at room temperature temperature cultivated on Hagem agar. After the fungus showed visible growth, a few pieces of mycelium were punched out and crushed with a Hagem liquid medium in a Buhler homogenizer (Edmund Buhler, Tübingen, G). The further culture then takes place in 250 ml Erlenmeyer flasks with Hagem liquid medium according to H. Kreisel and F. Schauer (methods of the ecological laboratory, Fischer-Verlag Jena 1987). In each case 100 ml of culture medium were mixed with 10 ml of the vaccine suspension prepared as described above. The cultivation was carried out at room temperature and constant light conditions for 30 days, with continuous mixing using a shaking machine (Innova 2100, New Brunswick Scientific Co, INC. EDISON, New Jersey, USA) being achieved at a speed of 125 rpm. The further scale-up was carried out by transferring it to a bioreactor of 10 1 volume. 6 1 autoclave HAGEM medium (liquid) was added to the fermenter. This was mixed with 60 ml vaccine suspension. The cultivation in the fermenter was carried out at room temperature with constant gassing with sterile air, continuous stirring and in a day-night rhythm. To determine the mycelium dry weight, 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.

All extracts were concentrated in a vacuum rotary evaporator (Büchi Labortechnik AG, Flawil, Switzerland) at 40 ° C. and then lyophilized (Gefriertrocknungsanlagen GmbH, Osterode, G). Result: The cultivation of G. pfeifferi is possible with liquid medium. The yield is on average 5 g mycelium / 1.

A cultivation in a 10 1 fermenter allows the production of mycelium in larger quantities.

Example 3 Methodology: Carrying out the experiments according to Example 2, but with the addition of 0.1-1%

Potassium thiocyanate.

Result: increase in the yield of mycelium from 0.34 + 0.0568 g / 100 ml to 0.691 +

0.048 g / 100 ml by adding 0.5% potassium thiocyanate.

Example 4: Extracts obtained by extracting the fruiting body or mycelium with a lipophilic solvent

Methodology: Lyophilized mycelium was filled into extraction tubes and extracted with various lipophilic solvents in a Soxhlet apparatus for 24 hours. All extracts were concentrated in a vacuum rotary evaporator (Büchi Labortechnik AG, Flawil, Switzerland) at 40 ° C, lyophilized (Gefriertrocknungsanlagen GmbH, Osterode, G) and then that

Dry weight determined.

Result: 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

Methodology: 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.

Result: Yields between 0.1 and 0.5% were obtained. The extracts obtained from fungal cultures without SCN addition showed an inhibitory effect against S. aureus with inhibitory areas up to 18 and 20 mm in the agar diffusion test, those from fungal cultures with SCN addition of 23 and 20 mm.

Tab. 1 Inhibitory effect against S. aureus

Example 6: Extraction of culture mycelium with water

Methodology: Culture mycelia according to Examples 2 and 3 were extracted with water at 70 ° C., in a further experiment first with water at 20 ° C. and then with water at 70 ° C.

Results: 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

Table 3: Recipe of the - Shii-take— micro- and nanoparticles

The biomass is heated to a temperature of 50 ° C. Separately, an aqueous emulsifier solution is heated to the appropriate temperature (50 ° C). Then both phases are combined at the desired homogenization temperature. Then 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.

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.

Table 4: Shii-take (Lentinula edodes) recipe - micro- and nanoparticles

5 g of vitamin C were then incorporated into the biomass. 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 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. The two micro and nanoparticles B30 and vitamin C were mixed together.

Example 9 Production of Micro and Nanoparticles from the Fruit Body of the Mushroom Judas Ear (Auricularia auricula-judae The 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.

Table 5: Prescription of the Judas ear (Auricularia auricula-judae) mushrooms - micro and nanoparticles

Example 10 Production of Micro and Nanoparticles from the Fruit Body of the Mushroom Ganoderma Lucidum - (Glossy Lacquer Porling

Fruiting bodies of the Ganoderma lucidum mushrooms were used.

Table 6: Recipe of the Ganoderma lucidum mushrooms - particles

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.

Example 11: Production of micro and nanoparticles from Mvcel of the Maitake mushroom (Grifola frondosa)

The mycelium of the mushroom Maitakev (Grifola frondosa) was processed into micro and nanoparticles.

Table 7: Recipe of the Nostocales - micro and nanoparticles

Example 12: Production of micro- and nanoparticles from fruiting bodies of Mai-take (Grifola frondosa with vitamin C according to the solvent method

Fruit bodies of the Maitake (Grifola frondosa) mushroom were used.

Table 8: (Solvent process)

First of all, 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.

Example 13 Production of Micro and Nanoparticles from Fruit Bodies of the Maitake Mushroom - Norlichexanthon

Table 9: Recipe of the biomass, northern exanthone, micro and nanoparticles

The biomass is heated to a temperature of 50 ° C. and then the marine active ingredient norlichexanthone 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.

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.

Table 10: Recipe for biomass, vitamin E, micro and nanoparticles

Example 15 Production of Micro- and Nanoparticles from Fruit Bodies of the Mushroom May-take (Grifola frondosa) - Provitamin Q10

Table 11 Recipe for Biomass-Pro vitamin Q10 micro and nanoparticles

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.

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)

[0080] The two substances were mixed together 1: 1.

Methodology:

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 μl 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 μl 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.

In the controls, only bacterial counts> 100 were found. From this, the mean number of germs n and the scatter were calculated.

In the examination of the preparations, bacterial counts between 0 and 100 were found. From this the mean number of bacteria m was calculated on the basis of the Poisson distribution according to the following formula: m = In (number of samples without detection of bacteria / total number of samples).

Statistical evaluation: The number of skin areas with and without germ detection for the control without treatment and after treatment with the test product was checked for significance using the chi-square test. Results

The testing of the micro- and nanoparticles, produced according to Example 8, brought about a marked reduction in the transmitted contamination with MRSA on the skin model (cow udder teat). The number of repetitions on the cow udder teat was 163 for the untreated controls. S. aureus was detectable in all 163 samples. The mean bacterial count ... S. aureus was also detected in all samples that were treated with wool wax alcohol ointment. After application of the test product, 11 skin areas with proof of germs and 11 without proof of germs were found. From this, an average bacterial count of 0.7 is to be expected (Fig. 5). The bacterial count reduction is cantly high sig- ^'. Example 17: Testing of micro- and nanoparticles from Mvcel of Shii-take mushrooms / vitamin C "produced according to Examples 7 and 8 on the animal model mouse ear

methodology

[0086] Mouse ears were used as the test model. The donor animals as a source of infection remained untreated. Accepted animals were treated for 3 days once a day with “Shii-take mushrooms / vitamin C” prepared in accordance with Example 8. 10 μl of test substance were pipetted on and ground with a glass spatula and dried at room temperature The donor animals were contaminated with 5 μl 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.

After incubation at 30 ° C for 1.5 hours, the skin areas of the acceptor ears were spread on Müller-Hinton plates and incubated at 37 ° C.

Evaluation: The evaluation and statistical backup was carried out as described in Example 9.

Results:

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 by means of skin contact was simulated. The number of repetitions on the mouse ear was 163 for the untreated controls, germ growth was detected on skin areas. In pretreatment with the preparation according to Examples 7 and 8, 12 skin areas with and 5 without germ detection. This means that after a corresponding pretreatment, only 1.2 germs / cm ² can be expected on average (Fig. 6).

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 μl 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.

Result

Despite the skin disinfection carried out, numerous coagulase-negative apathogenic staphylococci of the normal skin flora of the pig were detectable on the smear. The coagulase-positive aureus strains with which the contamination had been carried out were detectable in the untreated control in addition to S. epidermidis; in the animals treated according to the invention, all colonies examined were coagulase-negative.

Example 19: Testing of micro and nanoparticles from Mvcel of Shii-take mushroom / vitamin C produced according to Example 8 on the skin model according to Example 17

Result

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 microparticles 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 ^{2 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 (Grifolina 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 0.18 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

Results

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.

Result

The number of preparations tested with the Norlichexanthon was 22. The number of skin areas with germ detection was 19 and without germ detection 3. On average, 2 germs are expected after pretreatment with preparations according to the invention according to Example 14.

Example 22: Preparation of ubiquinone Q1 biomass particles

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. Tab. 12 Recipe of the ubiquinone biomass particles

Example 23: Prevention of the transmission of MRSA in skin contact with the preparation according to Example 22

methodology

Tails of aseptic mice were used for the experiments. In order to rule out 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.

The 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 smeared on blood Müller-Hinton aga plates. After incubating the Agaφlatten at 37 ° C for 24 h, the colonies were counted.

Results:

When the acceptors are streaked out immediately after skin contact, the plates are completely covered (number of germs> 10,000). This applies equally to pre-treated and to Controls without pre-treatment. This means that a massive transmission of MRSA can be simulated well in this model. The same number of bacteria is transmitted in the test and control groups.

2 h and 24 h, on the other hand, only low bacterial counts can be detected in the pretreated actuators, while the bacterial counts in the untreated controls remain high. (Tab. 1). With this test model, too, the interruption of the infection routes can be demonstrated by pretreatment with the formulations according to the invention.

Tab. 13 Germ counts on mouse tails (acceptors) depending on the pretreatment after contact with MRSA colonized mouse tails (donors).

These tests were carried out with other vitamins and antioxidant agents. Reductions were observed both in the mouse ear animal model and in the cow udder teat model in the case of the combination of norlichexanthon and pro vitamin Q10. However, the greatest germ reduction of approach germs was achieved with the combination of vitamin E, norlichexanthone and pro vitamin Q10. Example 24 Particle size determination of the micro and nanoparticles, produced according to Example 8

Methodology The particle size by photon correlation spectroscopy was determined using a Zetasizer III (Malvern, UK).

Example 25: Detection of the radical scavenging property of the extracts by means of chemiluminescence

The detection was carried out as follows:

A: 30 ml of human blood were preincubated with 170 ml of PBS + luminol (0.33 mM final) at 37 ° C. for 20 min.

B: 30 μl of human blood were preincubated with 170 ml of PBS + luminol (0.33 mM final) and 20 ml of substance dilution at 37 ° C. for 20 min. [Olli] To A, a 1: 100 dilution of the extract according to Examples B and C and 20 μl zymosan (10 mg / ml) were pipetted into A. 20 μl of zymosan (10 mg / ml) were pipetted into B.

As control, we used PBS instead of zymosan as a stimulator with and without substance in batches A and B. After intensive mixing, the kinetics of the luminescence was measured over 60 min. The tests were carried out on two different days with blood from two different blood donors.

Result: The extracts clearly inhibit the luminol-induced and the spontaneous oxygen radical release or intercept the released radicals. This effect is enhanced by the addition of thiocyanate.

Example 26: Testing the radical trapping properties of fungi using the α. α-Diphenyl-ß-picrylhydrazyl radical scavening effect (DPPH assay)

The DPPH assay was used to test the radical scavenger properties of the mushroom / vitamin combinations. principle The DPPH radical (2,2-diphenyl-l-picrylhydrazyl) has its absorption maximum at 517 nm and a violet coloration, 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 (2,2-diphenyl-ß-picrylhydrazyl) is formed. The absorption is also reduced, which can be measured with a photometer.

method

The comparison substance is dissolved in ethanol and the following dilutions are made: 10 μg / ml, 50 μg / ml, 100 μg / ml, 500 μg / ml.

ImM DPPH dissolved in ethanol corresponds to 294 μg / ml. 4 mg of the mushroom / vitamin combinations are dissolved in 4 ml of ethanol and the following dilution is made: 10 μg / ml, 50 μg / ml, 100 μg / ml, 500 μg / ml. Test procedure:

500 μl of the sample are pipetted into an Eppendorf vessel. 375μl ethanol and 125 μl DPPH solution are added. The solution is shaken and incubated for 30 min at room temperature in the dark. After this time, the absorption at 517 nm is measured in the photometer. The measured absorptions are then expressed in% radical scavenger activity in comparison to the blank value (875 μg / ml ethanol + 125 μg / ml DPPH.

100

Quantity in% = 100 - [4 sample control

Tab. 14 Result Auricularia auricula-judae - Judas ear mycelium / ascorbic acid

Tab. 15 Result Ganoderma lucidum - shiny lacquered porcelain mycelium / ascorbic acid

Tab. 16 Result Grifola frondosa - Maitake mycelium / ascorbic acid

Tab. 17 Result Hericium erinaceus - porcupine mycelium / ascorbic acid

Tab. 18 Result Lentinula edodes - Shii-take-mycelium / ascorbic acid

Example 27: Inhibition of neutral endopeptidase

Methodology: The detection of the specific inhibition of the neutral endopeptidase was carried out according to Melzig et. al., Pharmacy 51 (1996) 501-503.

Result: The extract of the mycelium according to Example B with 80% alcohol results in an inhibition of 68% at a concentration of 100 μg / ml and 84% at 200 μg / ml. In the corresponding ethanolic extract of the culture mycelium with SCN addition according to Example C, the activity of the enzyme was inhibited by 70% even at a concentration of 50 μg / ml.

[0123] The aqueous extracts from the mycelium are also effective. After incubation with 50 μg / ml of the cold-water extract, an inhibition of the enzyme activity by 65% was found. An influence of the SCN ^" could not be recognized.

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 μg / 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 trapping properties, provides protection against free radical damage to the skin.

Furthermore, 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 Mvcel of G. pfeifferi as vitalizing. pain-relieving and germ-reducing nutritional supplements

Methodology: The protection of the cells against toxic radicals, demonstrated according to Example E, can also be of nutritional significance 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. On the other hand, the specific inhibitory effect of the extracts from G. pfeifferi on the neutral endopeptidase can be used for pain relief, since after they interfere in the sense of inhibiting the enkephalinase with the breakdown of neuropeptides and endoφhins that attack the opiate receptors.

Result: The combination of radical scavenger properties, immunostimulating effect, pain relieving effect and properties preserving against microbial spoilage creates very favorable conditions for use as a nutritional supplement.

Example 31: Use as a health care product and nutritional supplement

Methodology: Aqueous and alcoholic extracts as well as nanoparticles from mushrooms of the genus Ganoderma obtained from the biomass inhibit the cholesterol accumulation in cultivated human aortic intimal cells. By inhibiting 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. 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 metabolic 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 0.1 and 0.2%.

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 count and abnormal widening of the vessels in the periphery of the body.

Legend to the figures

Figure 1: Growth curves of the biomass Ganoderma Pfeifferi in the SCN-containing medium

Figure 2: Growth curves of the biomass Ganoderma resinaceum in the SCN-containing medium

Figure 3 growth curves of the biomass Ganoderma applanatum in the SCN-containing medium

Figure 4: Growth curves of the biomass Ganoderma adspesum 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.

Figure 6: Evidence 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

Claims

claims

1. Compositions consisting of biomasses of terrestrial fungi on the one hand and inorganic thiocyanates or thiocyanates of organic bases on the other.

2. Compositions according to claim 1, characterized in that additionally contain one or more active ingredients.

3. Compositions according to claim 2, characterized in that they additionally contain one or more minerals and / or radical scavengers and / or nutritional supplements and / or vitamins, in particular vitamin C.

4. Compositions according to one of claims 1 to 3, characterized in that they additionally contain one or more dispersion-stabilizing substances.

5. Compositions according to one of claims 1 to 4, characterized in that as lipid-containing terrestrial mushrooms

a) Auricularia auricula-judae - Judas ear b) Ganoderma lucidum - glossy lacquered porling c) Grifola frondosa - Maitake d) Hericium erinaceus - porcupine ect.) Lentinula edodes - Shii-take.

6. Compositions according to one of claims 1 to 5, characterized in that the biomasses are converted into fungi into micro- and nanoparticles.

7. Compositions according to claim 6, characterized in that the micro- and nanoparticles have an average diameter of 10 nm - 10 microns.

8. A process for the preparation of compositions according to claim 1, characterized in that a) the cultivation of the mushrooms in the presence of inorganic thiocyanates or thiocyanates of organic bases or b) the biomass of the mycelium and / or the fruiting body of the mushrooms inorganic thiocy- anates or thiocyanates of organic bases can be added.

9. A process for the preparation of compositions according to claim 2 or 3, characterized in that a) the cultivation of the fungi takes place in the presence of inorganic thiocyanates or thiocyanates of organic bases and an enrichment with active ingredients by additives to the culture medium during the cultivation or b) the biomass of the mycelium and / or the fruiting body of the mushrooms, inorganic thiocyanates or thiocyanates of organic bases, and one or more active ingredients are additionally added.

10. The method for producing compositions according to claim 6, characterized in that the biomasses or the cultivated biomasses are converted into micro- and nanoparticles with a diameter of 10 nm - 10 μm by homogenization or emulsion formation.

11. The method according to claim 10, characterized by the following steps:

Heating the biomass mushrooms until the lipids contained therein liquefy, if necessary adding one or more active ingredients or additives

Mixing the biomass or the loaded biomass with a surfactant-water mixture heated to above the melting temperature of the fatty acids and combining the two phases. Establishing a pre-suspension high pressure homogenization in one or more homogenization cycles

12. The method according to claim 11, characterized in that the heating of the microorganisms and the surfactant-water mixture is omitted and the active ingredients are adsorbed at room temperature on the lipid-containing fungi or dispersed with the addition of a small amount of water.

13. The method according to claim 10, characterized by the following steps:

Suspend the biomass of the mushrooms and possibly the additives in an organic solvent and predisperse this mixture High pressure homogenization and subsequent spray or freeze drying

Redispersion in an aqueous surfactant solution, redispersion and high pressure homogenization in one or more

homogenization

14. The method according to claim 10, characterized by the following steps:

Emulsion formation from water and biomass and, if necessary, with the additives. Dissolving the emulsion in a suitable organic solvent. Adding a water-soluble co-surfactant and predispersion. High pressure homogenization and removal of the solvent.

15. Use of compositions according to at least one of claims 1 to 7 as active ingredient carriers.

16. Use of compositions according to at least one of claims 1 to 7 as health-promoting agents.

17. Use of compositions according to at least one of claims 1 to 7 for the production of health-promoting agents.

18. Use of compositions according to claims 1 to 7 for therapeutic and prophylactic applications in humans and / or animals.

19. Use of the compositions according to claims 1 to 7 as

a) Food b) Feed c) Food Supplements d) Feed Supplements

20. Use according to claim 19 as a nutritional feed.

21. Use according to claim 18 and 19 for improving the rearing results and for the prophylaxis and therapy of infectious diseases in animal husbandry and animal breeding

22. Use according to claim 21 in combination with other medicaments.

23. Use according to claim 15 as an antibiotic carrier.

24. Use of compositions according to claims 1 to 7 in the form of oils, sprays and / ointments and in capsules.

25. Use of compositions according to claims 1 to 7 for the targeted substitution of deficiency states.

26. Use of compositions according to claims 1 to 7 for immunostimulation.

27. Use of compositions according to claims 1 to 7 for the targeted substitution of the thiocyanate deficiency in dialysis patients.