EP2320869A1

EP2320869A1 - Surfactant-free emulsions with anti-aging effect, production thereof and use of same

Info

Publication number: EP2320869A1
Application number: EP09781131A
Authority: EP
Inventors: Wolf-Dieter Juelich; Hans-Peter Welzel; Ulrike Lindequist; Ottmar Geiger
Original assignee: HC Berlin Pharma Ag; Ernst Moritz Arndt Universitaet Greifswald
Current assignee: HC Berlin Pharma Ag; Universitaet Greifswald
Priority date: 2008-07-26
Filing date: 2009-07-27
Publication date: 2011-05-18
Also published as: DE102008035443A1; WO2010012686A1

Abstract

The invention relates to a method for producing surfactant-free emulsions from the annual mugwort (Artemisia annua) and fungi of the genus ganoderma, which are characterised by improved properties, especially free-radical scavenger properties and an anti-aging effect, and can therefore have multiple uses in cosmetics.

Description

Surfactant-free emulsions with anti-aging effect, their preparation and their use

The object of the invention is a process for the preparation of surfactant-free emulsions from the annual mugwort (Artemisia annua) and fungi of the genus Ganoderma, which are characterized by improved properties, in particular by free radical scavenging properties and an anti-aging effect and therefore diverse can be used in cosmetics.

State of the art

1. Solid state stabilized emulsions

The production of solid-state stabilized emulsions has been known for over 100 years.

2. Previous use of Artemisia annua

The annual mugwort {Artemisia annua) has been used for millennia as a medicinal plant. The plant is native to Europe and Asia. It grows 2 to 3 m high and has fresh green, heavily divided leaves and tiny cream-colored flower heads. For medical purposes, the aerial parts of the plant are used. The preparation and use of extracts from Artemisia and their use in the fields of nutrition, cosmetics and medicine is known from the literature (eg Applied Biochemistry and Biotechnology, 1990, VoI 24/25, pp 213-222).

The ingredients of the annual mugwort {Artemisia annua) have a high activity against the malaria parasite. In contrast to other malaria drugs, the drug has few side effects and also a very favorable resistance, whereby it can be combined with other substances.

The actual active ingredient is artemisinin (artemesin, cotexin), a secondary plant substance (sesquiterpene with peroxide group), which occurs in the leaves and flowers of annual mugwort {Artemisia annua). Like all phytochemicals, these are associated in the biomass with other similar substances, since the biochemical pathways are not single-tracked. So are 9 other related substances, which also against the Malaria parasites act in the plants. Over 200 ingredients could be detected in the plant parts in total.

The mode of action of artemisinin discussed in the literature is closely related to the chemical structure. The peroxide group contained in the molecule is unstable in the presence of high concentrations of iron ions and forms free radicals. Such high levels are found in red blood cells and also in malaria pathogens that accumulate iron. If artemisinin gets into erythrocytes, free radicals are formed and the parasite may be killed. There is also evidence that artemisinin derivatives inhibit certain enzymes.

From the artemisinin or related plant substances, several semi-synthetic derivatives have been prepared, but all must contain the Peroxidgruppierung to the efficacy.

There are first indications of an effect on various human cancers. According to results of the German Cancer Research Center Heidelberg, this effect is also based on the formation of free radicals by the peroxide grouping in the presence of iron ions. The examination of artemisinin as a potential anticancer drug is still at an early stage.

Further applications of the peroxide-containing structures on the skin, for example for local wound treatment, are described in US2007269537.

It is expected that the general demand for artemisinin products will increase to several hundred million treatments in the next few years. To produce this volume, enormous amounts of the herbal active ingredient are needed. It would therefore be of great advantage if there were the highest possible utilization of the resulting in large excess biomass.

From one hectare of plants can harvest one to two tons of leaves from which about two to three kilograms of artemisinin can be obtained. Extraction from vegetable raw materials is complicated by the fact that the artemisinin content of the plants is very low and has considerable fluctuations. It is usually between 0.1 and 0.4%, based on the dry weight. Ideally, can be from a hectare Plant material to harvest one to two tons of leaves from which by extraction with non-polar solvents such as hexane and subsequent treatment with petroleum ether about 2 to 3 kg of artemisinin can be obtained. The result is an oily-yellow extract of the plant, which is refined to gel and then to white, crystalline powder containing artemisinin as a so-called Blutschizontozid.

The remaining biomass remains largely ignored in the previous utilization concepts, since the use of the plant so far focused on the use of artemisinin as an antimalarial agent.

In contrast to Artemisinin relatively few applications for the other ingredients of the species Artemisia have been described.

3. Potential combination partners for the preparation of surfactant-free emulsions from Artemisia annua

Artemisia annua is mainly used in combinations. For example, KR20030051517 describes a dietary supplement containing, in addition to Hovenia dulcis Thunberg and Semisulcospira libertine, 8% of Artemisia luayomogi as its main constituents.

In KR20030005127 a dietary supplement is described, which in addition to Hoving Dulis Thunb and Alnus rubra Hovenia dulcis contains as main ingredients an addition of 10% Artemisia capillaris. Artemisia capillaris is described in CN1615927 among other things as an anti-hypertensive agent, the prior art is a mixture with various Chinese plants. CN 1969679 describes the combined use of Artemisia capillaris and G. lucidum in the form of a tea. In JP2000143437 a cosmetic is described which contains at least 2 of the following plant extracts: Veronica undulata Wall, Ottelia alismoides pers., Artemisia apiacea Hance, Artemisia annua L., Andrographis paniculata Nees, Dichroa febrifuga Lour., Eclipta prostrata L., Dipsacus asper Wall, Dipsacus japonicus Miq., Boehmeria nivea Gaud., Polygonum aviculare L., Sterculia lychnophera Hance, Carpesium abrotanoides L. and Polyporus mylittae Cook. A combination of sugar alcohols, such as xylitol or erythritol, and plant extracts, which may also contain, among others, Artemisia capillaries, is described in JP2001081008 for external application to the skin. A dietary supplement containing an Artemisia capillaris Thunb. Extract is described in KR20050024920. This nutritional supplement is said to counteract skin aging.

Lindenquist: Ganoderma In: Hagers Handbook of Pharmaceutical Practice / Ed. F. Bruchhausen, 5th completely revised edition, Springer- Verlag Berlin- Heidelberg New York 1998, Volume 2 Drogen AK (Ed. W. Blaschek), pp. 750-761 The beneficial ingredients of the genus Ganoderma are often combined with extracts from Artemisia spp. The combination described in KR20020078314 contains eg 3% Artemisia capillaris Thunb. And 2% G. lucidum Also described in JP2006143711 is a combination of extracts of plants and fungi which also contains, inter alia, Artemisia argyi Levl., Et Vant and G. lucidum The foodstuff described in KR20040032288 contains Artemisia capillaris Thunb in combination with brewer's yeast and G. lucidum.

KR20050001730 describes a dietary supplement with immuno-activating and anti-tumor activity, which i.a. G. lucidum (FR) Karst and Artemisia Messerschmidtiana Besser contains.

Triterpenes isolated from Ganoderma concinna can induce apoptosis in HL-60 cells (Gonzalez AG, Leon F, Rivera A, Padron JI, Gonzalez-Plata J, Zuluaga JC et al., New Lanostanoids from the Fungus Ganoderma concinna J Nat Prod 2002; 65: 417 '-2I). Triterpenes, available from Ganoderma applanatum, are effective against mouse skin tumors (Chairul, Tokuyama T, Hayashi Y, Nishizawa M, Tokuda H, Chairul SM, Hayashi Y. Applanoxidic acids A, B, C and D, biologically active tetracyclic triterpenes from Ganoderma applanatum, Phytochemistry 1991; 30: 4105-9; Chairul, Chairul SM, Hayashi Y. Lanostanoid triterpenes from Ganoderma applanatum, Phytochemistry 1994; 35: 1305-8). A derivative of illudin was effective in clinical studies (Murgo A, Cannon DJ, Blatner G, Cheson BD, Clinical Trials of MGI-114, Oncology 1999, 13: 233-8).

The administration of lentinan in addition to chemotherapy in patients with gastric cancer, colon cancer and other tumors has proved to be successful (Hazama S, Oka M, Yoshino S, Iizuka N, Wadamori K, Yamamoto et al., Clinical effects and immunological analysis of intraabdominal and intrapleural injection of lentinan for malignant ascites and pleural effusion of gastric carcinoma., Cancer & Chemotherapy 1995; 22: 1595-7). CN1351886 describes an agent of 20 components of Chinese medicine, which contains, inter alia, G. lucidum and Artemisia capillaris and which is to be used in liver diseases. CN1092295 also teaches the use of G. lucidum and Artemisia capillaris in a combination of many herbal extracts to combat various

Diseases.

JP2048517 describes a hair care product, which i.a. Artemisia apiacea Hance and

G. lucidum contains.

As can be seen from this list, although different species of the genus Artemisia are used in the described combinations of extracts of the genera Artemisia and Ganoderma, while the use of Ganoderma species has hitherto been limited to the use of the species G. lucidum. A combination of Artemisia annua with Ganoderma pfeifferi is not yet known, although extracts from G. pfeifferi from the German patent application DE 10 2005 031 363 Al are known. All applications described in the prior art use extracts from Artemisia annua. Extracts use only some of the ingredients. The ratio of the ingredients to each other is changed by the extraction process and no longer corresponds to the physiological conditions.

The use of triterpenes from Ganoderma species for the production of solid-stabilized emulsions has not been described. Also surfactant-free dermatic bases with anti-aging effect based on a combination of Artemisia annua and Ganoderma spec. are not known yet. In particular, neither Artemisia annua nor Ganoderma spec. containing spray emulsions based on solids-stabilized emulsions used for UV protection, although especially sunscreen and personal care sprays have established themselves in the market and are perceived by the user due to their ease of use as pleasant.

4. State of the art in intended applications anti-aging and sunscreen

Skin aging refers to the complex biological process of alteration of the skin associated with aging. While the intrinsic aging, ie the genetically controlled diminished reactivity of skin cells can not be influenced, extrinsic factors (environmental factors such as ultraviolet light, chemical light) can be influenced by extrinsic factors Reagents, mechanical stress, stress, heat and cold) caused cell aging can be reduced by anti-aging preparations. In particular, under the influence of free radicals, there is an exhaustion of the cell processes, cell division, increased permeability of the cell membranes and an insufficient supply of the cells.

As a visible sign of this environmental cell aging, the skin gets deep wrinkles and wrinkles, their dry surface tends to tears and pseudo colors, the epidermis is thinner. It is produced less fat, the skin loses its elasticity and is no longer so regenerative. Because UVA radiation penetrates deep into the skin, it produces singlet oxygen in the dermis. This causes the production of enzymes that damage the collagen fibers and thus reduce the firmness of the skin. At the same time, elastic fibers swell, resulting in loss of elasticity of the skin.

The problem of all cosmetic preparations in connection with anti-aging, however, is that so far, especially the visible external signs of aging are judged. The state of the art creams can only reduce these visible signs, as long as the cause - the environmental cell aging - is not eliminated. Existing wrinkles can not be made to disappear but essentially moisturise and moisturize the skin so that it temporarily appears smoother. Since the influence on the environmental aging of the cells could not be detected experimentally, the aging process of skin cells by the prior art corresponding means is practically not affected.

A major cause of premature skin aging is UV radiation. In conventional sunscreens based on titanium dioxide (TiO ₂ ), the reflection and adsorption by microfine particles of titanium dioxide or zinc oxide, which reflect the incident UV light, exploited to avoid erythema (sunburn). Act as inorganic UV filters. In modern preparations, the pigment particles are reduced to about 200 nanometers.

When exposed to ultraviolet (UV) light, photocatalytic substances such as titanium dioxide (TiO ₂ ;) highly absorb UV radiation. In the reaction of titanium dioxide with UV radiation, however, free radicals are formed in the presence of water and oxygen. It has been proven that by the photocatalytic effect of Titanium dioxide in addition to premature cell aging when using titanium dioxide in UV protection also damage the DNA caused by free radicals whose formation by the protective agent does not prevent, but is even enhanced as a result of the photocatalytic reaction.

Virtually no sunscreen therefore today dispenses with the addition of radical scavengers. This makes sense because reactive oxygen species are involved in all inflammatory processes and, above all, attack the unsaturated compounds (amino acids, proteins, lipids) that make up the cell walls and DNA structures of the cell nuclei. Free radicals and reactive oxygen species also play a role in polymorphic photodermatosis - often referred to by the layman as a sun allergy. In sunscreen products there are therefore a large number of substances which are intended to neutralize free radicals: vitamin E, vitamin C, glucosylrutin, follicular glucitol, ginkgo extract, thermal water, silymarin, superoxide dismutase, green tea extract, bacterial lysates, organic melanin, ferulic acid or carboxymethyl glucan. For some substances, however, it is questionable whether they also act as free-radical scavengers when applied topically. A potent radical scavenger is the flavonoid glucosylrutin, which is applied in combination with vitamin E as a Pre Sun cream for the prevention of polymorphic light dermatosis days before staying in the sun (Hadschiew 1997). The same principle follows the recommendation to saturate the skin early with a highly concentrated vitamin E cream (example Optolind E) (Heinrich 1994).

Almost every sunscreen contains vitamin E (tocopherol). Pure vitamin E creams achieve a sun protection factor of about 3. The peroral intake in the epidermis can not achieve sufficiently high tocopherol concentrations. In addition, studies have shown that exposure to sunlight can reduce skin vitamin E levels by up to fifty percent (Thiele 1998). Therefore, a local application is required. As an acetate, vitamin E penetrates well into the epidermis, where it is cleaved by esterases into free vitamin E. Due to its structure, it can be stored well in the cell wall and protects it from the attack of radicals. To characterize the quality of a sunscreen protection against sunburn in the future will no longer be in the foreground. Although the SPF helps to prevent erythema when used properly, it does not provide guidance on how the consumer can avoid immunosuppression or irreparable nuclear damage. This requires new measurement criteria. In the photocatalytic reaction, very reactive free OH radicals are formed which have a strong antimicrobial action (A. Heller: Chemistry and Applications of Photocatalytic Oxidation of Thin Organic Films Acc. Chem. Res., Vol 12 (1995) 503 / D. Bahnemann: Photocatalytic Detoxification of Polluted Waters.The Handbook of Environmental Chemistry, Springer Verlag 1999, Volume 2, Part L, 285-351. It becomes a very strong reduction under photocatalytic activation by UV light the initial germ content of E.faecium is 0.01%, that of S. aureus is 0.001%, and that of E. coli is 0.00002%, and this strong biocidal activity is undesirable in skin applications.

Object of the invention

It was therefore an object of the invention to develop new surfactant-free skin care products, on the one hand to avoid the disadvantages mentioned and on the other hand allow new applications of the biomass of Artemisia annua as a cosmetic or pharmaceutical agents.

Presentation of the invention

The object is achieved according to the features of the claims, according to the invention by a pharmaceutical or cosmetic composition comprising extracts of plants of the species Artemisia annua and extracts of fungi of the genus Ganoderma, wherein the extracts of plants of the species Artemisia annua obtained with polar solvents after removal of artemisinin (sesquiterpene peroxides) in a nonpolar solvent extraction step.

The sub-task to find applications for those ingredients of Artemisia annua, which do not carry peroxide grouping and therefore are not suitable for malaria therapy, according to the invention, on the one hand by a two- or three-stage extraction method and on the other hand solved by suitable combinations.

In the multistage extraction process according to the invention is extracted in the first step with nonpolar solvents. By extraction with pentane, hexane, Petro lether or other non-polar solvents, the removal of artemisinin and lipids, which can be fed to a separate use. The extraction method with nonpolar Solvents such as pentane or petroleum ether can be carried out in a known manner both batchwise and with continuous processes such as extraction by means of Soxhlet apparatus or percolator. The resulting plant material is suitably freed from adhering residual solvent, for example by vacuum drying.

In the context of this invention, it was found that the resulting biomass - in the presence of iron ions - in contrast to the starting material has no cytotoxicity. It is therefore much better suited as a component in cosmetics than the direct use of the plant material corresponding in the prior art. The resulting biomass contains active ingredients with a high radical scavenging capacity, which protect the plant from self-damage by free radicals. To utilize this radical scavenger capacity, the resulting biomass is subjected to a second extraction according to the invention. This, in turn, may be alcoholic, aqueous-alcoholic or aqueous, surprisingly finding active substances with free radical scavenging properties in each extract.

In the three-stage extraction according to the invention first artemisinin and lipids are removed from the plant material as in the two-stage process with nonpolar solvents such as pentane, hexane or petroleum ether.

The extracted plant material is freed, for example by vacuum drying, of the adhering residual solvent and subjected to a second extraction with lower alcohols such as methanol or ethanol, resulting in an alcoholic extract. In a third extraction stage can now after removal of residual alcohol, for. B. in vacuo or even without removal of the still adhering alcohol, other agents are obtained by aqueous extraction.

As extraction method for the alcoholic extraction, both discontinuous and continuous methods such as extraction by means of Soxhlet apparatus or percolator are suitable. The aqueous-alcoholic or aqueous extraction is expediently carried out discontinuously, for example in the form of a maceration.

In a particularly advantageous manner, the object of improving the applicability of the biomass of Artemisia annua as a health-promoting agent, was solved by either dried and crushed plants of the species Artemisia annua or the residues the above-described extraction of the plants with pentane, hexane, petroleum ether or other non-polar solvents on the one hand with dried and crushed mycelium and / or fruiting bodies of fungi of one or more species of the genus Ganoderma on the other hand mixed and the mixture of an alcoholic, aqueous-alcoholic or aqueous extraction is subjected.

It is also inventive to carry out the multi-stage extraction of the dried and shredded plants of the species Artemisia annua and fungi of the genus Ganoderma separately and to mix the various extracts obtained in a suitable ratio.

The high triperpene content of the selected species of the genus Ganoderma forms the prerequisite for the preparation of surfactant-free emulsion systems as a basis for innovative dermatics and cosmetics. Triterpenes are biologically active phytochemicals that have antimicrobial, especially antiviral and anti-inflammatory properties. Surprisingly, the content of some triterpenes in G. pfeifferi is partially increased up to a factor of 100 compared with G. lucidum. The content of the triterpene Lucialdehyd B in G. lucidum is, for example, 0.14 mg / 100 g, in G. pfeifferi, however, 19.2 mg / 100 g.

To produce surfactant-free emulsion systems, the plant and mushroom extracts obtained according to the invention, if appropriate with addition of further triterpenes, in particular of triterpenic acids such as ursolic acids, betulinic acid and / or boswellic acids, are suspended in natural waxes, preferably in jojoba oil. Homogenising gives a solid-stabilized emulsion with an anti-aging effect, which makes it possible to produce various cosmetic products without further emulsifiers, especially without synthetic surfactants. In accordance with the invention, micropigments incorporated as solids, preferably metal oxides and / or other sparingly soluble or insoluble metal compounds in water, as well as possibly also the UV filters - in addition to their actual purpose - also contribute to the stabilization of the formulation. The micropigments bind to the oil phase of the system and thereby simultaneously improve the spreadability of the formulation on the skin. Due to the stabilizing effect of the micropigments can be largely dispensed with other surface-active excipients. This reduces the risk of skin intolerance altogether. In surfactant-free sunscreen emulsions eliminates possible interactions of surfactants with the skin. As relatively small surfactant molecules, surfactants tend to interact in particular with the intercellular lipids of the stratum corneum and thus can weaken the epidermal barrier function. In addition, due to their penetration-promoting properties, they are able to channel ingredients of the formulation with irritative or allergenic potential more strongly into the skin. It has long been known that the combination of UVA radiation and certain ingredients in cosmetics, especially lipids and surfactants, can trigger the so-called Mallorcan acne. When using the inventive preparations which are free of synthetic surfactants and at the same time have a high UVA protection, this typical skin reaction is not expected. The jojoba oil contained in the preparations according to the invention enhances the cytoprotective effect and reduces the risk of side effects.

A further advantage of surfactant-free emulsion systems is that they are very suitable as bases for spray products. Surfactant-containing spray emulsions feel sticky on the skin. In contrast, the inventive solid-stabilized emulsions protect the skin without leaving a greasy film on the skin. In particular, for use as a sunscreen agent, it is advantageous to additionally stabilize the spray emulsions according to the invention by adding copolymers of the polyvinylpyrrolidone. Polymers based on polyvinylpyrrolidone (PVP) or other polymeric film formers, such as sodium polystyrene sulfonate, form after evaporation of the solvent, a film that contributes significantly to the protective function.

The sunscreen according to the invention act prophylactically by the combination of UV filters, radical scavengers and anti-inflammatory substances in preventing UV-related premature skin aging and, moreover, anti-inflammatory.

In higher concentrations, the extracts of Artemisia annua have a high radical scavenger capacity, but significantly decreases at higher dilutions. Surprisingly, it has been found that even small additions of extracts, for example from G. pfeifferi, cause a strong increase in free-radical scavenging capacity. For example, the aqueous extract from the combination Artemisia annua: Ganoderma pfeifferi = 99: 1 had a radical scavenging activity of 89% in a dilution of 1 to 100 in the DPPH test. Without the addition of G. pfeifferi, the radical scavenging activity of Artemisia annua in this dilution is not sufficient (radical scavenging activity of the aqueous or alcoholic extract in the DPPH test between 34 and 38%). The combination with G. pfeifferi thus additionally reduces the oxidative stress and enhances the endogenous radical defense and thus supports the anti-aging effect. The ingredients ganomycin A and B, which are only found in G. pfeifferi, inhibit luminol-induced and spontaneous oxygen radical release at a concentration of 10 μg / ml. The combination increases the proliferation of human keratinocytes. Under the influence of the combination, the glucose consumption of human cells increases, indicating a stimulation of the metabolism. It has been found that the formation of proteins is promoted and cell aging is slowed down.

The increased cell permeability under UV influence - measurable by determining the LDH activity in the medium - is reduced. The regeneration capacity of the cells after UV damage is significantly increased.

In the photocatalytic reaction, u.a. very reactive free radicals that have a strong antimicrobial effect. Accordingly, the physiological skin flora is severely damaged. The combination according to the invention greatly reduces this effect.

In the combination of Artemisia annua and Ganoderma spec. Both in the joint extraction of biomass and in the combination of the extracts virtually all mixing ratios are possible. It can be used advantageously also mixtures of different types of Ganoderma. In an advantageous embodiment of the invention, extracts of different types of Ganoderma are mixed. In particular, it is advantageous to mix the aqueous extract of G. lucidum with the ethanolic extract of G. pfeifferi. The aqueous extract of G. lucidum contains agents that induce apoptosis of tumor cells, activate phagocytosis by macrophages and stimulate the immune response, the alcoholic extract of G. pfeifferi contains active ingredients that stimulate the metabolism of healthy cells. The stimulating effect on healthy cells is only detectable in lipophilic extracts of G.pfeifferi. However, the glucan content of the aqueous extract, which stimulates the immune system, is significantly lower in G.pfeifferi than in G. lucidum or G. tsugae. The combination of the ethanolic extract of G. pfeifferi and the aqueous extract of G. lucidum or G. tsugae produces the desired effect.

A significant advantage of the preparations according to the invention is that the cell protection is completed by triterpenes from Ganoderma species. In particular, triterpenes from Ganoderma concinna can initiate apoptosis of cells at an early stage which has degenerated despite the radical scavenging effect. Tetracyclic triterpenes and lanostanoid triterpenes from Ganoderma applanatum support the body in the defense against tumors, so that their admixture brings particular benefits.

Active ingredients derived from the fruiting body and the mycelium of the genus Ganoderma not only supplement the action of the ingredients of Artemisia annua as radical scavengers, but additionally bring about an inhibition of the activity of neutral endopeptidases and / or an inhibition of the activity of the angiotensin converting enzyme , The cosmetic and pharmaceutical preparations according to the invention may contain further plant extracts. Particularly suitable are the extracts of special plants such as green tea, witch hazel, chamomile, marigold, Paeonie, aloe vera, horse chestnut, sage, willow bark, hawthorn, lime blossom, almonds, spruce needles, sandalwood, coconut, kiwi, guava, lime, mango, apricot , Wheat, melon, orange, grapefruit, avocado, rosemary, mallow, meadowfoam, thyme, lemon balm, marshmallow, mallow, ginseng, birch, with extracts from the divisible forming tissue being preferred. Furthermore, it may be preferable to use in the compositions according to the invention mixtures of several different plant extracts.

The cosmetic and pharmaceutical preparations according to the invention can also be combined with algae extracts. Suitable algae extracts come from green algae, brown algae, red algae or blue algae (cyanobacteria).

It has been found that the formulations according to the invention, which use the natural substances of various plants and fungi, make better use of their active ingredients by the solid-state-stabilized emulsions according to the invention and bring them to penetration more rapidly than formulations with large drops of oil.

Sprays based on the surfactant-free emulsions according to the invention can be used both for protection against UV radiation and as after-sun products. Preparations for protection against UV radiation in the context of the present invention preferably contain at least one UV-A, UV-B and / or broadband feed substance. The formulations may, although not necessary, optionally also contain one or more organic and / or inorganic pigments as UV filter substances, which may be present in the water and / or the oil phase. The pigments can be used advantageously in the context of the present invention also in the form of commercially available oily or aqueous Predispersions are used. The pigments may advantageously be surface-treated (coated), forming, for example, a hydrophilic, amphiphilic or hydrophobic character, This surface treatment may consist in using a thin hydrophilic and / or hydrophobic pigment inorganic and / or organic layer The various surface coatings contribute according to the invention to stabilize the emulsion.

Inorganic surface coatings in the context of the present invention may consist of aluminum oxide (Al 2 O 3), aluminum hydroxide Al (OH) ₃ or aluminum oxide hydrate (also: alumina, CAS No .: 1333-84-2), sodium hexametaphosphate (NaPOs) ₆ , Sodium metaphosphate (NaPO ₃ ) ", silicon dioxide (SiO ₂ ) (also: silica, CAS No .: 7631-86-9), or iron oxide (Fe ₂ O ₃ ). These inorganic surface coatings may be present alone, in combination and / or in combination with organic coating materials.

Organic surface coatings in the context of the present invention may consist of vegetable or animal aluminum stearate, vegetable or animal stearic acid, lauric acid, dimethylpolysiloxane (also: dimethicone), methylpolysiloxane (methicone), simethicone (a mixture of dimethylpolysiloxane with an average chain length of 200 to 350 dimethylsiloxane units and silica gel) or alginic acid. These organic surface coatings may be present alone, in combination and / or in combination with inorganic coating materials.

The list of said UV filters, which can be used in the context of the present invention, should of course not be limiting. Advantageously, the preparations of the invention contain the substances which absorb UV radiation in the UV-A and / or UV-B range, in a total amount of z. 0.1% by weight to 30% by weight, preferably 0.5% to 20% by weight, in particular 1.0% to 15.0% by weight, based in each case on the total weight of the preparations, of cosmetic preparations available which protect the skin from the entire range of ultraviolet radiation.

After-sun products and anti-aging products based on the surfactant-free emulsions according to the invention may contain further active ingredients. In order to increase the protective effect, preferably vitamins and antioxidants are selected from the group consisting of amino acids (eg glycine, histidine, tyrosine, tryptophan) and their derivatives, imidazoles, (eg urocaninic acid) and their derivatives, peptides such as D, L-carnosine, D-carnosine, L-carnosine and their derivatives (eg anserine), Carotenoids, carotenes and their derivatives, chlorogenic acid and its derivatives, lipoic acid and its derivatives, aurothioglucose, propylthiouracil and other thiols (eg thioredoxin, glutathione, cysteine, cystine, cystamine and their glycosyl, N-acetyl, methyl, ethyl, Propyl, amyl, butyl and lauryl, palmitoyl, oleyl, γ-linoleyl, cholesteryl and glyceryl esters) and their salts, diauryl thiodipropionate, distearyl thiodipropionate, thiodipropionic acid and derivatives thereof (esters, ethers, peptides, lipids, nucleotides , Nucleosides and salts) and sulfoximine compounds (eg buthionine sulfoximines, homocysteine sulfoximine, buthione sulfones, penta, hexa, heptathionine sulfoximine) in very low tolerated dosages (eg pmol to μmol / kg), furthermore (metal) chelators, Hydroxy acids (eg citric acid, lactic acid, malic acid), humic acid, bile acid, bile extracts, bilirubin, biliverdin, EDTA, EGTA and their derivatives, unsaturated fatty acids and their derivatives, vitamin C and derivatives (eg ascorbyl palmitate, magnesium ascorbyl phosphate, ascorbyl acetate), tocopherols and Derivatives (eg vitamin E acetate), vitamin A and derivatives (eg vitamin A palmitate) and benzylic resin, rutinic acid and derivatives thereof, ferulic acid and its derivatives, furfurylidene glucitol, carnosine, butylhydroxytoluene, butylhydroxyanisole, nordohydroguajaretic acid,

Trihydroxybutyrophenone, quercitin, uric acid and its derivatives, mannose and its derivatives, selenium and its derivatives (e.g., selenium methionine), stilbenes and their derivatives (e.g., stilbene oxide, trans-stilbene oxide). Mixtures of antioxidants are also suitable for use in the cosmetic preparations according to the invention. Particular preference is given to phenolic acids and flavonoids.

Furthermore, the inventive preparations to support the anti-aging effect protein hydrolysates or their derivative may contain. According to the invention, both vegetable and animal protein hydrolysates can be used. Animal protein hydrolysates are z. B. elastin, collagen, keratin, silk and milk protein protein hydrolysates, which may also be in the form of salts. Vegetable protein hydrolysates, eg. Soy, wheat, almonds, peas, potato and rice protein hydrolysates. Although the use of the additional protein hydrolysates is preferred as such, amino acid mixtures or individual amino acids obtained otherwise, for example, may optionally be present in their place Arginine, lysine, histidine or pyrroglutamic acid. Also possible is the use of derivatives of protein hydrolysates, z. In the form of their fatty acid condensation products. In the cosmetic and pharmaceutical preparations according to the invention, the additional protein hydrolysates and their derivatives are contained in amounts of 0.01 to 10% by weight, based on the total agent. Amounts of 0, 1 to 5 wt.%, In particular 0, 1 to 3 wt .-%, are particularly preferred.

According to the invention contain anti-aging preparations for the care of the skin moistening or moisturizing agents (so-called moisturizers). The surfactant-free emulsions are therefore advantageously substances selected from the group of glycerol, lactic acid and / or lactates, in particular sodium lactate, butylene glycol, propylene glycol, Biosaccaride Gum-1, Glycine soybean, Ethy lhexy Io xy glycerol, pyrrolidonecarboxylic acid and urea added. Furthermore, it is particularly advantageous to use polymeric moisturizers from the group of water-soluble and / or water-swellable and / or water-gellable polysaccharides. The preparations according to the invention may contain mono-, oligo- or polysaccharides or their derivatives. Suitable monosaccharides are, for. As glucose, fructose, galactose, arabinose, ribose, xylose, lyxose, allose, altrose, mannose, gulose, idose and talose, the deoxy sugars fucose and rhamnose and amino sugars such. As glucosamine or galactosamine. Preferred are glucose, fructose, galactose, arabinose and fucose; Glucose is particularly preferred.

Suitable oligosaccharides are composed of two to ten monosaccharide units, e.g. As sucrose, lactose or trehalose. A particularly preferred oligosaccharide is sucrose. Also particularly preferred is the use of honey, which contains predominantly glucose and sucrose.

Suitable polysaccharides are composed of more than ten monosaccharide units. Preferred polysaccharides are the starches made from aD-glucose units and starch degradation products such as amylose, amylopectin and dextrins. Particularly advantageous according to the invention are chemically and / or thermally modified starches, for. B. Hydroxypropylstärkephosphat, Dihydroxypropyidistärkephosphat or further preferred are dextrans and their derivatives, for. B. dextran sulfate. Also preferred are nonionic cellulose derivatives such as methylcellulose, hydroxypropylcellulose or hydroxyethylcellulose, as well as cationic cellulose derivatives. Further preferred examples are polysaccharides from fucose units. Especially preferred are the polysaccharides composed of amino sugar units, in particular chitins and their deacetylated derivatives, the chitosans, and mucopolysaccharides. The inventively preferred mucopolysaccharides include hyaluronic acid and its derivatives, e.g. As sodium hyaluronate or Dimethylsilanolhyaluronat, and chondroitin and its derivatives, for. B. chondroitin sulfate.

The polymeric active ingredients used in surfactant-free systems do not penetrate the skin due to their molecular size, but remain as a film on the skin surface. Of course, it is known to the person skilled in the art that cosmetic preparations are generally not conceivable without further customary auxiliaries and additives. Accordingly, the cosmetic and dermatological preparations may, for example, comprise bodying agents, fillers, perfumes, foaming inhibitors, dyes, pigments, coloring matter, thickening agents, insect repellents, water, salts, proteolytically or keratolytically active substances, medicaments or other conventional ingredients of a cosmetic or dermatological formulation. Thus, with the surfactant-free emulsions innovative products for cosmetics and health care products can be made available with a new quality. The inventive separation of the peroxide-containing compounds before use as a cosmetic care properties are improved. In addition, it is not insignificant that the production of special Ganoderma species, which have not yet been used commercially on a larger scale, is very complicated due to the slow growth of fungi. The combination partner from Artemisia annua, on the other hand, is a by-product in the production of the antimalarial drug. With the combination according to the invention of special Ganoderma species and Artemisia annua residues, therefore, a cost-effective possibility for the production of an anti-aging active substance with the cell metabolism-stimulating properties is made available.

A major advantage of surfactant-free spray emulsions is that they can be autoclaved after preparation. This allows a germ-free application on the skin without the need for a preservative. Contamination during use is largely excluded by the spray form.

If in other applications of the inventive preparations are not dispensed preservative, antimicrobial agents are selected from the Groups of parabens, such as methylparaben, ethylparaben, propylparaben, isopropylparaben, butylparaben, isobutylparaben, sodium-methylparaben, sodium-ethylparaben, sodium-propylparaben, sodiumisobutylparaben, sodiumisopropylparaben or sodium-butylparaben, imidazolidinyl urea, diazolidinyl urea, iodopropynyl butylcarbamate, 2-bromo- 2-nitropropane-1,3-diol, sorbic acid, potassium sorbate, cetyltrimethylammonium chloride, cetylpyridinium chloride, benzethonium chloride,

Diisobutylethoxyethyldimethylbenzylammonium chloride, diisobutylphenoxyethoxyethyldimethylbenzylammonium chloride, N-alkyl-N, N-dimethylbenzylammonium chloride, bromide, saccharinate, trimethylammonium chloride, sodium aluminum chlorohydroxylactate, triethyl citrate, tricetylmethylammonium chloride, tri-chloro-trichloro -l'-hydroxydiphenyl ether (triclosan), 3,4,4'-trichlorocarbanilide (triclocarban), diaminoalkylamide, for example, L-lysine hexadecylamide suitable.

Terpenes and / or antimicrobial agents can be released from the preparations according to the invention in certain applications. This is particularly important for the following specific application.

The human pathogenic species of the genus Candida colonize skin and mucous membranes in many healthy people, without causing symptoms. However, if the fungal cells proliferate, eg after damage to the bacterial mucous membrane flora by antibiotics or with reduced immune defense, they often cause local inflammations, which are also referred to as thrush. As more and more sensitive textiles, such. As silk or microfiber, which can be washed only at 30 or 40 ⁰ C, are processed into garments, fungi such as Candida albicans are not killed. In particular, after a fungal infection it can come so by adhering to the garments, not killed fungi to a reinfection. To prevent the infection by fungi and / or to prevent the attachment of fungi to surfaces so far antimicrobial substances are used, which either inhibit the growth of fungi (fungistats), kill them (fungicides) and / or reduce the adhesion of the fungi. Frequently non-selective antimicrobial substances are used which act both against bacteria and against fungi. In addition, compounds are often used which, due to the concentration in which they must be used, have toxic properties, can cause skin irritation or have other undesirable properties. Surprisingly, it has now been found that the emulsions according to the invention in suitable formulations release terpenes and / or antifungal agents, are able to very effectively reduce the adhesion of microorganisms to surfaces, wherein the surfaces are both natural surfaces as well as artificial surfaces and in this case both biotic and abiotic surfaces. For this particular embodiment, the terpenes to be released are selected from terpene alcohols, ie such terpenes, preferably mono-, sesqui- and / or diterpenes, which carry a free hydroxyl group. Citronellols, geraniol, farnesol and patchouli alcohol are particularly preferred. According to a further preferred embodiment, perfume alcohols selected from eugenol, cinnamyl alcohol and anethole, in particular eugenol, can be added to the emulsions in this specific application.

2,5-Dihydroxybenzoic acid, which was found in G. pfeifferi, but not in other Ganoderma species, acts as antifungal agent. The antiadhesive substances are hereby preferably present in amounts of up to 3% by weight, more preferably in amounts of from 0.1 to 1% by weight, in the preparations. For certain applications, the combination of the antimicrobial peroxide-containing active substances from Artemisia annua with the biomass of G. pfeifferi derived antimicrobial agents, in particular with the ganomycins, with 2,4,5-trihydroxybenzaldehyde and / or 2,5-dihydroxybenzoic acid of particular advantage.

An advantage of the emulsions according to the invention is that they have only very low toxicity when maintaining these concentrations, in particular in comparison to the conventionally used antimycotics, fungicides and fungistats.

The invention will be explained in more detail by the following examples, without being limited thereto:

embodiments

Example 1 Production of Biomass of the Species Ganoderma pfeifferi

Methodology: Freshly harvested young fruiting bodies of the species G. pfeifferi were purified and disrupted under sterile conditions. From the area of the hat-to-pedicle transition, tissue pieces were removed with sterile forceps and cultured at room temperature on Hagem agar. After the fungus showed visible growth, some mycelial pieces were punched out and minced with a Hagem liquid medium in the Bühler homogenizer (Edmund Bühler, Tübingen, G). Subsequently, the further culture in 250 ml Erlenmeyer flask with Hagem liquid medium after H. Kreisel and F. Schauer (methods of myko logical 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. Culturing was performed at room temperature and constant light for 30 days, with continuous mixing using a shaker (Innova 2100, New Brunswick Scientific Co., INC. EDISON, New Jersey, USA) at a speed of 125 rpm. The further scale-up was carried out by transfer into a bioreactor of 10 1 volume. The fermenter was treated with 6 l of autoclaved HAGEM medium (liquid). This was mixed with 60 ml of seed suspension. Cultivation in the fermenter was carried out at room temperature with continuous gassing with sterile air, continuous stirring and in the day-night rhythm. The mycelium was separated from the medium by filtration and lyophilized.

Result:

The cultivation of G. pfeifferi is possible with liquid medium. The yield is on average 5 g mycelium / 1. The lyophilized or air dried mycelium is available for blending with the biomass of Artemisia annua.

Example 2 Extraction of Extracts of the Species Ganoderma pfeifferi Methodology:

Ganoderma pfeiffer fruiting bodies were obtained by wild collection in Schlosspark Ludwigsburg. In addition, cultured fruiting bodies, which were obtained by the company GAMU GmbH, Krefeld, were used.

The collected fruiting bodies of Ganoderma pfeifferi were purified, cut into small pieces, dried in air at room temperature and then freeze-dried in a high vacuum. The cut and dried pieces were then pulverized in an analytical mill and then stored in opaque containers at room temperature. By means of three different extraction methods were gained ethanolic extracts. Applied were a hot or cold extraction and extraction with the extractor.

hot extraction

The pulverized fruit body was mixed in a paper tube with ethanol 80% (V / V) and then extracted by the Soxhlet method for about 24 h. The initial weight for G. pfeifferi was 12.5 g. The recovered liquid extracts were concentrated and freeze-dried.

cold extraction

For cold extraction, in each case 25 g of drug material were mixed with 250 ml of 80% ethanol (v / v) and shaken at room temperature for 24 h. The fungi were then squeezed through a filter, the resulting cloudy extract centrifuged and filtered through a round filter. The filtrates were concentrated and freeze-dried. The storage of the extracts obtained took place in the refrigerator.

Extraction by extractor

The extraction was carried out by means of extractor ASE 200. In each case 2 g of drug material were weighed with 15 g of sea sand in a paper tube. Extraction was carried out at a pressure of 2000 psi (pounds per square inch) with a service life (residence time of the solvent in the extraction cell at each extraction step) of 2 minutes. As extraction agent EtOH 80% (V / V) was used at an extraction temperature of 60 ⁰ C.

Result:

The yields of freeze-dried extracts were summarized in Table.

Tab. 1: Extract yields of G. pfeifferi u

The following terms were used in the following: GPS collected G. pfeifferi hot extract

GPK collected G. pfeifferi cold extract

GPKK cultured G. pfeifferi cold extract

GPE collected G. pfeifferi extract extract

Example 3 Extraction of total extracts from Artemisia annua (including artemisinin)

Example 3a: Alcoholic Extraction (discontinuous)

3 g of dried plant material Artemisia annua are mixed with 30 ml of methanol and allowed to stand for 12 hours at room temperature. Thereafter, the plant material is pressed and filtered through a Faltenfüter. This results in 20 ml of methanolic extract from Artemisia annua.

Example 3b: Alcoholic extraction (continuous, Soxhlet extraction)

10 g of dried plant material Artemisia annua are extracted in a Soxhlet apparatus continuously with 150 ml of methanol for 1 hour. This results in 130 ml of methanolic extract from Artemisia annua.

Example 3c: Aqueous-Alcoholic Extraction (discontinuous)

3 g of dried plant material Artemisia annua are mixed with 40 ml of 40% aqueous ethanol and allowed to stand for 12 hours at room temperature. Thereafter, the plant material is pressed and filtered through a pleated filter. This results in 32 ml of aqueous-ethano lischer extract from Artemisia annua.

Example 3d: Aqueous extraction (discontinuous)

3 g of dried plant material Artemisia annua are distilled with 30 ml. Water and allowed to stand for 12 hours at room temperature. Thereafter, the plant material is squeezed off and the cloudy solution is centrifuged. This results in 20 ml of aqueous extract from Artemisia annua.

Example 4 Extraction of Artemisia annua (artemisinin free) extracts Example 4a: Separation of the peroxide-containing compounds by extraction of Artemisia annua with petroleum ether (discontinuous)

3 g of dried plant material Artemisia annua are mixed with 30 ml of petroleum ether and allowed to stand for 12 hours at room temperature. It is then pressed off and filtered, resulting in 20 ml petroleum ether extract. The plant material is freed from adhering solvent residues in vacuo for 30 minutes.

Example 4b: Separation of the peroxide-containing compounds by extraction of Artemisia annua with petroleum ether (continuous)

10 g of dried plant material Artemisia annua are extracted in a Soxhlet apparatus continuously with 150 ml of petroleum ether for 1 hour. This results in 140 ml of extract from Artemisia annua. The remaining plant material is freed of the adhering solvent residues in vacuo.

Example 4c: Extraction of Peroxide-Free Extracts by Extraction with Methanol (Continuous)

[0104] Approx. 3 g of plant material from the previous extraction according to Example 4 a are extracted continuously in a Soxhlet apparatus with 100 ml of methanol for 1 hour. This results in 90 ml of peroxide-free methanolic extract.

Example 4d: Extraction of Peroxide-Free Extracts by Extraction with Ethanol (discontinuous)

[0105] Approx. 10 g of dried plant material Artemisia annua from the previous extraction according to Example 4 b are mixed with 100 ml of ethanol and allowed to stand for 12 hours at room temperature. Then it is pressed off and filtered. This results in 80 ml of peroxide-free ethanol extract.

Example 4e: Aqueous-Alcoholic Extraction (discontinuous)

10 g of dried plant material Artemisia annua are mixed with 80 ml of 40% aqueous ethanol and allowed to stand for 12 hours at room temperature. Thereafter, the plant material is pressed and filtered through a pleated filter. This results in 70 ml of aqueous-ethano lischer extract from Artemisia annua. Example 4f: Aqueous extraction (discontinuous)

Example 5 Obtaining the anti-aging agents according to the invention by co-extraction of Ganoderma pfeifferi and Artemisia annua. (Continuous)

1 g of the powdered fruit body of G. pfeifferi were mixed with 10 g of dried and by extraction of artemisinin-free plant material from Artemisia annua in a paper sleeve with methanol and then extracted by the Soxhlet method for about 2 h. The recovered liquid extracts were concentrated and freeze-dried. For use in cosmetic preparations, they may be used directly or in a solvent.

Example 6 Obtaining the anti-aging agents according to the invention by co-extraction of Ganoderma pfeifferi and Artemisia annua. (discontinuous)

1 g of the powdered fruit body of G. pfeifferi were mixed with 10 g of artemisinin-dried artemisinin-free plant material from Artemisia annua, added with 80 ml of 40% aqueous ethanol and allowed to stand for 12 hours at room temperature. Thereafter, the plant material was squeezed and filtered through a pleated filter. This resulted in 70 ml of aqueous ethanolic extract of Ganoderma pfeifferi and Artemisia annua.

Example 7 Obtaining the anti-aging agents according to the invention by mixing extracts of Ganoderma pfeifferi and Artemisia annua.

hot extraction

0.5 g of freeze-dried extract from G. pfeifferi was taken up in 100 ml of aqueous ethanolic extract from Artemisia annua (ethanol content 40%), giving a clear solution. The resulting total extract is directly in cosmetic

Preparations can be used. Example 8 Diphenylpikrylhydrazyl (DPPH) Test

[Olli] The determination of the antioxidative properties was carried out by thin-layer chromatography (SIEVERS A et al., (2002) Simple thin-layer chromatography test for antioxidative compounds using DPPH assay, CBS Camag Bibliography Service 88: 14-15) and photometrically (MOLYNEUX P (Mol. 2004) The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity: Songklanakarin J Sei Technol 26 (2): 211-219).

Results:

At higher concentrations (dilutions 1:20 or 1:10) Artemisia annua extracts show high free radical scavenging capacity (> 80%). However, in the 1: 100 dilution, the scavenging capacity is insufficient. Addition of 1% G. pfeifferi increases the free-radical scavenger capacity (Table 2).

Table 2:

Example 9: Detection of radical scavenging property by means of chemiluminescence

The detection was carried out as follows:

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

B: 30 μl of human blood was mixed with 170 ml of PBS + luminol (0.33 mM final) and 20 ml

Substance dilution preincubated at 37 ° C for 20 min.

To A, 20 μl of an extract containing G. pfeifferi at a 1: 100 dilution and 20 μl of zymosan (10 mg / ml) were pipetted.

To B was pipetted 20 μl of zymosan (10 mg / ml). As control we used PBS instead of zymosan as stimulator with and without substance in the beginnings A and B. After intensive mixing the kinetics of the luminescence over 60 min was measured. The tests were carried out on two different days with blood from two different blood donors.

Result:

The extracts clearly inhibit luminol-induced and spontaneous

Oxygen radical release or capture the released radicals.

Example 10 Detection of Intercellular Proteins Methodology

The following method was used to quantify the intercellular proteins. Of the

Detection of proteins with Roti - Nanoquant is based on a modification of the

Protein determination according to Bradford.

The Roti-Nanoquant working solution was prepared by diluting the stock solution in

Ratio 1: 5 with double-distilled water. produced. For this experiment was a culture bottle

(25 cm ² ), which was at least 80% confluent. The medium was decanted from the bottle and the cell lawn washed once with PBS. Subsequently, 50 μl of MMC

1 mg / ml in 5 ml of medium is added to the cells. The thus treated bottle was in

Incubator incubated at 37 ⁰ C, 95% humidity and 5% CO ₂ for 2 hours. To

Expiry of the incubation period, the mixture MMC / medium was poured off and the

Cell lawn washed twice with PBS. Subsequently, the cells were detached from the bottom of the bottle, the cell count determined and adjusted with nutrient medium to 200,000 cells / ml. 100 μl / well was placed in a 96-well plate. The plate was incubated in the incubator for 24 h.

After 24 hours, various extracts were pipetted into the cell culture plate.

The extracts of G. pfeifferi were used and combinations with

Artemisia annua.

For this purpose, a working solution of the corresponding extract was prepared. Of the

Dry extract was weighed exactly and distilled into 200 μl of ethanol. solved. From the batch, a stock solution of 1 mg / ml in double-distilled water. produced. The working solutions were prepared by diluting the stock solution with nutrient medium.

The old medium was removed and then 200 .mu.l / well of the various

Use solutions given to the plate. As a negative control were some wells with 200 μl DMEM (extract-free) filled. The thus treated plate was incubated for 24 h in the incubator.

In the next step, the destruction of the cell walls followed, in order to bring the intercellular proteins into solution and then quantify them. For this purpose, the use solutions were first removed from the plate and then washed 5 times with 200 ul / well PBS. After adding 100 .mu.l of a 0.1% Tritonlösung in Tris buffer (pH 7.2) per well, the plate was shaken for 30 min and then frozen. After thawing, the contents of the wells were thoroughly mixed using a pipette and then centrifuged at 4000 rpm for 4 minutes. From the plate thus treated, 10 μl each were transferred to the corresponding well of a new plate, the last row was released for the calibration curve. For this purpose, albumin, fraction V was used. A stock solution of concentration 1 mg / ml in 0.1% Triton solution in Tris buffer (pH 7.2) was prepared. Starting from this, the corresponding dilutions (0-500 μg / ml) were then prepared and 10 μl each pipetted into the corresponding wells. Subsequently, 40 μl of PBS and 200 μl of Roti-Nanoquant working solution per well were pipetted. After 5 min of incubation, the optical density (OD) was determined at 2 wavelengths (405 nm and 620 nm). For evaluation, the quotient of 620 nm and 405 nm was formed.

Results

The protein content increases under the influence of G. pfeifferi (GPS) and combinations containing G.pfeifferi to 110 and 125%. Already with a very small proportion of G. pfeifferi (5 μg / ml) an increase of the intercellular proteins compared to the negative control can be observed. An increase in the G. pfeifferi proportion in the mixture does not lead to an improvement in the effect.

Example 11: Preparation with aloe vera

methodology

Cells: FL, origin: USA, ATCC, CCL 62, catalog number RIE 81, supplier: biometec

GmbH

Medium: DMEM without phenol red with 100 μg / l glucose and 110 mg / l Na pyruvate (Gibco), modified with 20 mM L-glutamine, IM Hepes, 5 U penicillin / 5 mg streptomycin / ml (Sigma) and 10% FCS (Biochrom KG, Berlin) Elimination of cell division: First, a mitomycin C stock solution was prepared from 2 mg mitomycin C (Sigma) dissolved in 0.1 ml DMSO and 1.9 ml PBS and frozen in 50 μl aliquots. For mitosis inhibition, the cell culture medium was decanted from the confluent cell lawn of the FL cell culture, replaced with 5 ml of culture medium with 50 μl Mitomyxin stock solution and incubated for 2 hours / 37 ° C. Thereafter, it was washed 3 times with PBS (without Ca ⁺⁺ 'Mg ⁺⁺ , PAA), the cells were detached with trypsin / EDTA (Sigma), the cell count was determined and adjusted to a concentration of 4 × 10 ⁵ / ml with cell culture medium.

Cultivation on the cell carriers: In each cell suspension (13 mm round, Thermanox platelets (Nunc Ine, Naperville, IL USA) in a Minusheet cell holder system (Minucells & Minutissue GmbH, Bad Abbach) in a microculture well a 24-well Cell culture plate (Nunc GmbH & Co KG, Wiesbaden) pipetted, cultured for 24 hours (37 ⁰ C, 5% CO ₂ , 97% humidity) and then each set up 6 cell carriers in a perfusion cell chamber upright.

Perfusionszellkultur:

Cell chambers: Minucells & Minutissue, type 1301 conditioning at about 37 ⁰ C (hot plate: MEDAX, type 12501)

Flow rate: 4.5 μl / min (pump: Ismatec IPC-N 8)

Gas: low-gas type 110741 -S (5% carbon dioxide, 20% oxygen, balance nitrogen), Air

Liquide

Determination of the glucose concentration according to Beckmann (measuring intervals: 12 h +/- Ih), the pH and the oxygen partial pressure (measuring interval 24 h +/- Ih)

Determination of the minimum effective dose

11.1. Limitation of the application period

The addition of G. pfeifferi to the medium as a 2% suspension was limited in contrast to Example 3 to 12 h (from H 148 to h 160).

Result:

Already after this brief exposure time, a promoting effect can be seen, which is detectable for at least 3 days (FIG. 1). Fig. 1 shows an extension of Action time of the 2% suspension to 72 h increases the effect (Fig. 2), the promotional effect is detectable over a longer period.

11.2. Limitation of daily feed

To determine the minimum effective concentration, the daily intake was limited to 0.5 ml of the 2% suspension given within 111 min, i. Thereafter, the cells received over 1329 min a medium without drug addition. The daily delivery of the drug was reduced to 0.012% of the cell culture medium with this assay.

Even at this low concentration, a promoting effect can be seen (FIG. 3). When doubling the concentration (2 times a day for 111 min), the effect is more pronounced (Figure 4).

Claims

claims

Pharmaceutical or cosmetic composition comprising extracts of plants of the species Artemisia annua and extracts of fungi of the genus Ganoderma, characterized in that the extracts are obtained from plants of the species Artemisia annua with polar solvents, after in a previous extraction step with nonpolar solvents Artemisinin (sesquiterpene peroxides) was removed.

2. Composition according to claim 1, characterized in that it is extracts of fungi of the genus Ganoderma lucidum and / or Ganoderma pfeifferi and / Ganoderma applanatum and / or Ganoderma concinna and / or Ganoderma tsugae.

3. Composition according to claim 1 or 2, characterized in that surfactant-free, solids-stabilized emulsions are prepared by combining the extracts of Artemisia and Ganoderma and by the addition of further triterpenes and natural waxes.

4. Composition according to one of claims 1 to 3, characterized in that it additionally comprises the following ingredients:

4.1. further triterpenes and / or

4.2. natural waxes, preferably jojoba oil and / or

4.3. inorganic pigments, preferably metal oxides and / or other sparingly soluble or insoluble metal compounds in water and / or

4.4. Polymers based on polyvinylpyrrolidone (PVP) or other polymeric film formers and / or

4.5. known UV filters and / or

4.6. Vitamins and other antioxidants and / or

4.7. moisturizing or moisturizing agents and / or

4.8. known cosmetic adjuvants and / or

4.9. antimicrobial substances

5. Composition according to claim 4, characterized in that, as inorganic pigments, oxides of titanium (TiO ₂ ), zinc (ZnO), iron (eg Fe ₂ O), zirconium (ZrO ₂ ), silicon (SiO ₂ ), Manganese (eg MnO), aluminum (Al ₂ Os), Cers (eg Ce ₂ Os), mixed oxides of the corresponding metals and mixtures of such oxides and the sulfate of barium (BaSO ₄ ) are used.

6. A composition according to claim 4, characterized in that the UV filter used is 3- (4'-methylbenzylidene) -dl-camphor, 1- (4-tert-butylphenyl) -3- (4-methoxyphenyl) -propane

1, 3-dione, 4-isopropyldibenzoylmethane, 2-hydroxy-4-methoxybenzophenone, octyl methoxycinnamate, 3,3,5-trimethylcyclohexylsalicylate, 2-ethylhexyl 4- (dimethylamino) benzoate, 2-cyano-3,3-diphenylacrylic acid-2 - Ethylhexylester, 2-phenylbenzimidazole-5-sulfonic acid and their potassium, sodium and Triethanolaminsalze find use.

7. The composition according to claim 4, characterized in that as vitamins or other antioxidants are selected from a group consisting of amino acids (eg glycine, histidine, tyrosine, tryptophan) and their derivatives, imidazoles, (eg urocanic acid) and derivatives thereof, peptides such as D, L-carnosine, D-carnosine, L-carnosine and their derivatives (eg anserine), carotenoids, carotenes and their derivatives, chlorogenic acid and its derivatives, lipoic acid and its derivatives, aurothioglucose, propylthiouracil and other thiols (eg thioredoxin, glutathione , Cysteine, cystine, cystamine and their glycosyl, N-acetyl, methyl, ethyl, propyl, amyl, butyl and lauryl, palmitoyl, oleyl, γ-linoleyl, cholesteryl and glyceryl esters) as well as their salts, Diaurylthiodipropionat, Distearylthiodipropionat, thiodipropionic acid and its derivatives (esters, ethers, peptides, lipids, nucleotides, nucleosides and salts) and Sulfoximinverbindungen (eg Buthioninsulfoximine, Homocysteinsulfoximin, Buthioninsulf one, penta-, hexa-, heptathionine-sulfoximine) in very low tolerated dosages (eg pmol to μmol / kg), furthermore (metal) chelators, hydroxy acids (eg citric acid, lactic acid, malic acid), humic acid, bile acid, bile extracts, bilirubin , Biliverdin, EDTA, EGTA and their derivatives, unsaturated fatty acids and their derivatives, vitamin C and derivatives (eg ascorbyl palmitate, magnesium ascorbyl phosphate, ascorbyl acetate), tocopherols and derivatives (eg vitamin E acetate), vitamin A and derivatives (eg vitamin - A-palmitate) and coniferyl benzoate of benzoin, rutinic acid and derivatives thereof, Ferulic acid and its derivatives, furfurylidenglucitol, carnosine, butylhydroxytoluene, butylhydroxyanisole, nordohydroguajaretic acid, trihydroxybutyrophenone, quercitin, uric acid and its derivatives, mannose and its derivatives, selenium and its derivatives (eg selenium methionine), stilbenes and their derivatives (eg stilbene oxide, trans-stilbene oxide) ,

8. The composition according to claim 4, characterized in that the moisturizing or moisturizing or film-forming substances are selected from the group glycerol, lactic acid and / or lactates, in particular sodium lactate, butylene glycol, propylene glycol, Biosaccaride Gum-1, Glycine Soy, Ethylhexyloxyglycerin, Pyrrolidoncarbonsäure and Urea, or from the group of water-soluble and / or water-swellable and / or water-gellable polysaccharides, particularly advantageously hyaluronic acid and / or chitosan, polymers based on polyvinylpyrrolidone (PVP) or other polymeric film-forming agents, such as sodium polystyrene sulfonate

9. A process for preparing the composition according to any one of claims 1 to 8, characterized by the following steps: a) removing the artemisinin (sesquiterpene peroxides) by extraction with non-polar solvents from plants of the species Artemisia annua b) preparing an extract with alcoholic and aqueous alcoholic solvents from the residue of a) c) preparation of an extract with alcoholic or aqueous-alcoholic solvents from fungi of the genus Ganoderma d) preparation of an extract with aqueous solvents from the residue of a) e) preparation of an extract with aqueous solvents from the Residue of b) f) Preparation of an extract with aqueous solvents from the residue of c) g) mixing of the extracts obtained in step b) to e).

10. The method according to claim 10, characterized in that the steps b) and c) and the steps d), e) and f) take place in a reaction vessel and thus step g) is omitted.

11. The method according to claim 10 or 11, characterized in that polar, alcoholic, aqueous-alcoholic or aqueous extracts of the plants of the species Artemisia annua and alcoholic, aqueous-alcoholic or aqueous extracts of fungi of the genus Ganoderma are present, with different mixing ratios are possible

12. Anti-aging surfactant-free emulsions containing a composition according to any one of claims 1 to 9.

13. A surfactant-free sunscreen with anti-aging effect containing a composition according to any one of claims 1 to 9

14. Sprays containing surfactant-free preparations according to any one of claims 1 to 13.

15. Use of the composition according to one of claims 1 to 9 a) for skincare or b) for sun protection, in particular in the form of a spray or c) as an antimicrobial agent or d) as an antifungal agent or e) for the prophylaxis of skin tumors