WO2010012687A2 - Preparations based on artemisia annua, the conversion thereof into microparticles and nanoparticles, and use of same - Google Patents

Preparations based on artemisia annua, the conversion thereof into microparticles and nanoparticles, and use of same Download PDF

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Publication number
WO2010012687A2
WO2010012687A2 PCT/EP2009/059671 EP2009059671W WO2010012687A2 WO 2010012687 A2 WO2010012687 A2 WO 2010012687A2 EP 2009059671 W EP2009059671 W EP 2009059671W WO 2010012687 A2 WO2010012687 A2 WO 2010012687A2
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Prior art keywords
micro
nanoparticles
artemisia annua
characterized
ganoderma
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PCT/EP2009/059671
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German (de)
French (fr)
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WO2010012687A3 (en
Inventor
Wolf-Dieter Juelich
Hans-Peter Welzel
Ulrike Lindequist
Gerold Lukowski
Ottmar Geiger
Sabine Mundt
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Hc Berlin Pharma Ag
Ernst-Moritz-Arndt-Universität Greifswald
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Priority to DE102008035442A priority patent/DE102008035442A1/en
Application filed by Hc Berlin Pharma Ag, Ernst-Moritz-Arndt-Universität Greifswald filed Critical Hc Berlin Pharma Ag
Publication of WO2010012687A2 publication Critical patent/WO2010012687A2/en
Publication of WO2010012687A3 publication Critical patent/WO2010012687A3/en

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06Fungi, e.g. yeasts
    • A61K36/07Basidiomycota, e.g. Cryptococcus
    • A61K36/074Ganoderma
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/12Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/30Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K40/00Shaping or working-up of animal feeding-stuffs
    • A23K40/30Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A23B - A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L31/00Edible extracts or preparations of fungi; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A23B - A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23PSHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00Shaping or working of foodstuffs characterised by the products
    • A23P10/30Encapsulation of particles, e.g. foodstuff additives
    • A23P10/35Encapsulation of particles, e.g. foodstuff additives with oils, lipids, monoglycerides or diglycerides
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06Fungi, e.g. yeasts
    • A61K36/07Basidiomycota, e.g. Cryptococcus
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K8/00Cosmetics or similar toilet preparations
    • A61K8/02Cosmetics or similar toilet preparations characterised by special physical form
    • A61K8/11Encapsulated compositions
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K8/00Cosmetics or similar toilet preparations
    • A61K8/18Cosmetics or similar toilet preparations characterised by the composition
    • A61K8/96Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K8/00Cosmetics or similar toilet preparations
    • A61K8/18Cosmetics or similar toilet preparations characterised by the composition
    • A61K8/96Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9706Algae
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILET PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/41Particular ingredients further characterized by their size
    • A61K2800/412Microsized, i.e. having sizes between 0.1 and 100 microns

Abstract

The invention relates to preparations based on Artemisia annua, and to a plurality of methods for producing microparticles and nanoparticles from the annual mugwort (Artemisia annua). Ultrafine particles with improved properties are obtained as a result, that can be used as food supplements and feed supplements, in cosmetics and for medical purposes.

Description

Preparations of Artemisia annua, their conversion into micro- and nanoparticles and their use

[0001] The subject matter of the invention are preparations of Artemisia annua, as well as several processes for the production of micro- and nanoparticles from the sweet wormwood (Artemisia annua). As a result, ultrafine particles are obtained with improved properties that can be used as food and feed supplements for use in cosmetic, and medical applications.

State of the art

1. Preparation of nanoparticles and microparticles

[0002] For the preparation of nanoparticles and microparticles are already known various manufacturing processes on the basis of lipids or polymers. Thus, the process for the preparation of lipid microparticles have already been described based on phospholipids, which have anti-fungal properties and can be used in pharmaceutical or cosmetic field (DE 69 00 2905 T2). Other methods describe lipid nanoparticles based on extracted mono-, di- and triglycerides, oils or waxes. With the help of these lipids obtained pharmaceutical active ingredients are also encapsulated (WO 94/20072 Al). Other lipid nanoparticles also describe substances based on lipids for parenteral administration (WO 98/56362 Al). Also, special techniques for the production of lipid nanoparticles are introduced (eg. For example, EP 0526666 A). However, no micro and nanoparticles based on Artemisia annua and their preparation have been described in the literature.

[0003] The preparation of microparticles and nanoparticles by using species-specific lipids is already known (WO2004 / 075907 A2, and WO2003 / 72118 Al).

[0004] The use of the plant Artemisia annua focused so far on the use of artemisinin as an antimalarial agent. This requires the extraction with apolar solvents such as n-hexane. Here, the lipids are removed inevitably. A direct use of the teaching of WO2004 / 075907 A2 and WO2003 / 72118 Al for the conversion of the remaining biomass of Artemisia annua, which contains other valuable ingredients in micro and nano therefore is not possible because of the separated lipids. [0005] The residual biomass and thus the active ingredients contained therein remain largely ignored in previous valuation concepts.

2. Previous use of Artemisia annua

[0006] The Annual mugwort (Artemisia annua) is used as a medicinal plant for thousands of years. The plant is native to Europe and Asia. It grows 2 to 3 m high and has fresh green, strongly divided leaves and tiny cream-colored flower heads. For medicinal purposes, the aerial parts of the plant are used. The production and use of extracts of Artemisia and their application in the fields of nutrition, cosmetics and medicine is known from the literature (eg. As Applied Biochemistry and Biotechnology, 1990, VoI 24/25, pp 213-222).

[0007] A portion of the ingredients of the annuals, mugwort (Artemisia annua) has a high activity against malaria pathogens. Unlike other anti-malarial drugs, the drug no side effects and also a very favorable resistance profile, and it may also be combined with other substances.

[0008] The actual active substance is the artemisinin (Artemesin, Cotexin), a secondary plant substance (sesquiterpene peroxide), which occurs in the leaves and flowers of annuals mugwort (Artemisia annua). Like all plant compounds they are associated in the biomass with other similar substances, as the biochemical pathways do not run a single track. So are 9 other peroxide-containing substances which may also act against the malaria parasite, contained in the plants. Over 200 ingredients could be detected in the plant parts as a whole.

[0009] The discussion in the literature of action of artemisinin is closely related to the chemical structure. The peroxide group contained in the molecule is unstable in presence of high concentrations of iron ions and forms free radicals. Such high concentrations are found in red blood cells and also in malaria parasites that accumulate iron. Artemisinin arrives in erythrocytes, free radicals are formed, and the parasite may be killed by it. But there is also evidence that artemisinin derivatives inhibit certain enzymes. [0010] several semi-synthetic derivatives were prepared from the artemisinin or related plant substances, but must include all the efficacy which Peroxidgruppierung.

[0011] Further applications of the peroxide-containing structures are described in US2007269537, for example, for local treatment of wounds.

[0012] It is assumed that the overall demand for artemisinin drugs will increase in the years to several hundred million treatments. To produce this volume, enormous amounts of herbal drug are needed. It would therefore be of great advantage if there was a quality as possible exploitation of the resulting in large excess biomass.

[0013] From one hectare of plants can harvest one to two tons of leaves, of which about two to three kilograms of artemisinin can be obtained. The extraction of vegetable raw materials is made difficult by that the artemisinin content of the plants is very low and has considerable variation. It is typically between 0.1 and 0.4%, based on the dry weight. Ideally, can be harvested from one hectare Pfianzenmaterial one to two tons of leaves, from which approximately 2 to 3 kg of artemisinin can be obtained by extraction with nonpolar solvents such as hexane and subsequent treatment with petroleum ether. There arises thereby a yellow oily extract of the plant, which is refined into gel, and then to white, crystalline powder containing artemisinin as a so-called Blutschizontozid.

[0014] The residual biomass is largely ignored in the previous valuation concepts, since the use of the plant so far focused on the use of artemisinin as an antimalarial agent. To solve the problem posed suitable combination partners must be found.

3. Marine organisms as potential combination partners

[0015] For the task suitable natural products have been found for example in marine microalgae (Lukowski G Lindequist U, Mundt S, Jülich WD. Pharmaceutically or cosmetically active agent from lipid-containing marine organisms (2003) WO2003 / 72118 Al). Because marine organisms have very specific lipid compositions, it is possible to adjust formulations produced by selection of marine organisms from each of a particular use (Lindequist U, Schweder T: Marine biotechnologically Io gy. In: Rehm HJ. Biotechnologically Io gy second ed, Wiley-VCH Weinheim 2001. 442-473).

[0016] In addition, cyanobacteria have some remarkable biosynthetic potencies. Haematococcus pluvialis is for example a unicellular green alga that can produce the ketocarotenoid astaxanthin up to 5% of their biomass. Astaxanthin is a natural antioxidant is a valuable cosmetic raw material. While cyanobacteria are known with diverse biological effects as highly productive producers of secondary materials, the eukaryotic microalgae are especially appreciated as producers of primary materials such as fatty acids, pigments, among others.

[0017] compounds which are biogenetisch assign the polyketides were far less often found in microalgae (Falch, B. Phaarmazie in our time in 1996, 25, 311-312; Burja, AM; Banaigs, B .; Abou-Mansour, E .; Burgess, JG; Wright, PC Tetrahedron 2001, 57, 9347-9377). The first Paracyclophanes, chloride-containing secondary metabolites of Polyketidstoffwechsels were at the beginning of the 90s from Nostoc linkia and Cylindrospermum isolated licheniforme and an inhibitory effect on the proliferation of KB and LoVo cell lines demonstrated (Moore, BS, Chen JL, Patterson GML; Moore, RE ; Brinen, L .; KatoY .; Clardy, JJ Am Chem Soc 1990, 112, 4061-4063;... Chen, JL; Moore, RE; Patterson, GMLJ Org Chem 1991, 56, 4360-4364;.. Moore , BS; Chen, JL; Patterson, GML; Moore RE Tetrahedron, 1992, 48, 3001-3006). From a naturally occurring Nostoc sp. Bloom is a further cyclophane that is characterized by a triple bond, and antibacterial and antialgal effects has isolated (Ploutno, A .; Carmeli, SJ Nat. Prod. 2000, 63, 1524-1526).

4. Medical used fungi as potential combination partners

[0018] The lipid content of fungi can vary from less than 1% up to 15- 20% of the dry weight. The agent can be expected about 2 to 8%. Mushroom fat contains all kinds of lipids such as free fatty acids (mostly 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 Add Edible mushrooms and Their cultivation, 27-40, CRC Press, Boca Raton (1989); Garcha, HS, Khanna, PK, Soni, GL. Nutritional importance of mushrooms Add: 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 .: technology of Processing of mushrooms, publishing Eugen Ulmer, Stuttgart 21-55, (1974). [0019] the Ganoderma (Ganoderma spec) are wood inhabitants (Saprobionten and parasites) on coniferous and deciduous trees, which, inter alia, by a high Triterpern content excel. Biologically active compounds from mushrooms n of the genus Ganoderma have long been described (U. Lindequist: Ganoderma. In: Hager's Manual of pharmacy practice / Ed F. of Bruchhausen, 5th completely revised edition, Springer-Verlag Berlin Heidelberg New York 1998 sequel 2 Drug AK (eds W.Blaschek). S. 750-761.

5. So far realized combinations with artesunate

[0020] In contrast to the use of artemisinin as a malaria drug relatively few additional applications of plants of the genus Artemisia are reported so far. [0021] Thus, in KR20030051517 a dietary supplement is described as main constituents contains an addition of 8% Artemisia luayomogi next Hovenia dulcis Thunberg and Semisulcospira libertine.

[0022] In KR20030005127 describes a dietary supplement containing as main constituents contains an additive of 10% Artemisia capillaris Thunb next Hoving Dulis and Alnus rubra Hovenia dulcis. Artemisia capillaris is described inter alia in CN1615927 as agents against hypertension, wherein the prior art, a mixture having various Chinese plants corresponds. CN 1969679 describes the combined use of Artemisia capillaris and G. lucidum in the form of a tea. [0023] In JP2000143437 is described a cosmetic containing at least two 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 that may contain Artemisia capillaries, among others, is described in JP2001081008 for external application on the skin. [0024] A dietary supplement containing a Artemisia capillaris Thunb. containing extract, is described in KR20050024920. These dietary supplements of aging is to counteract. Artemisia annua also mainly used in combinations. [0025] The health-promoting ingredients of the genus Ganoderma are often with extracts of Artemisia sp. combined. Thus, the combination described in of KR20020078314 contains, inter alia, Artemisia capillaris Thunb 3%. and 2% G. lucidum. Also in JP2006143711 a combination of extracts from plants and fungi is described including Artemisia argyi Levl. et Vant and G. lucidum contains. The food described in KR20040032288 containing Artemisia capillaris Thunb lucidum combined with brewer's yeast and G..

[0026] KR20050001730 describes a dietary supplement with immune-activating and anti-tumor effect, which, inter alia G. lucidum (FR) Karst and Artemisia Messerschmidtiana Better contains.

[0027] CN1351886 discloses a composition consisting of 20 components of Chinese medicine, which contains, inter alia, G. lucidum and Artemisia capillaris and to be used for liver diseases. Also CN 1092295 teaches the use of G. lucidum and Artemisia capillaris in a combination of many herbal extracts to combat various

Diseases.

[0028] JP2048517 describes a hair care product, among other things, the Artemisia apiacea Hance and

G. lucidum contains.

[0029] As is apparent from this list will be used namely different kinds of the genus Artemisia in the described combinations of extracts of the genus Artemisia and Ganoderma, while the use of Ganoderma species is so far largely 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 10 2005 031 363 Al are known from the German patent application DE.

6. State of the art for intended applications

6.1. Anti-Aging

[0030] the complex biological process associated with aging skin change is referred to as aging. While intrinsic aging so the genetically controlled reduced sensitivity of skin cells can not be influenced, can (by extrinsic factors environmental factors such as UV light, chemical

be reduced reagents, mechanical load, stress, heat and cold) caused by cell aging anti-aging preparations. Especially under the influence of free radicals leads to a depletion of cellular processes of cell division, to increased permeability of cell membranes and a reduced supply of cells.

[0031] As a visible sign of the environmental cell aging the skin gets deep folds and wrinkles, their dry surface is prone to tears and pseudo scars, the epidermis becomes thinner. It is produced less fat, the skin loses elasticity and is no longer capable of regeneration. Since mainly the 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 elastic fibers swell, resulting in a loss of elasticity of the skin.

[0032] The problem of all cosmetic products related to anti-aging, however, that so far above all the visible external signs of aging are assessed. The state of the art creams these visible signs can only mitigate, as long as the cause - the environmental cell aging - is not eliminated. Existing wrinkles can not be made to disappear, but out of the skin essentially moisture and fat, so it appears temporarily smoother.

[0033] Since the impact on the environmental aging of cells so far has not been demonstrated experimentally, the aging process of the skin cells is practically not affected by the state of the art means.

6.2. sun protection

[0034] A significant cause of premature skin aging is UV radiation. [0035] In conventional sunscreens on the basis of titanium dioxide (TiO 2) the reflection and absorption is exploited by micro-fine particles of titanium dioxide or zinc oxide, that reflect the incident ultraviolet light, in order to avoid formation of erythema (sunburn). act as inorganic UV filters. In modern preparations, the pigment particles are reduced to about 200 nanometers.

[0036] For exposure to ultraviolet (UV) light absorbing photocatalytic substances such as titanium dioxide (TiO 2) to a high degree of UV radiation. In the reaction of titanium dioxide with UV radiation, however, form in the presence of water and oxygen free radicals. It is demonstrated that also damage to the DNA caused by the photocatalytic effect by titanium dioxide with the use of titanium dioxide in UV protection agents in addition to a premature cellular aging caused by free radicals, their formation is not prevented by the protective means, but is even increased, due to the photocatalytic reaction.

[0037] Virtually no sunscreen therefore waived today the addition of free radical scavengers. This is quite useful because reactive oxygen species are involved in all inflammatory processes and attack mainly the unsaturated compounds (amino acids, proteins, lipids) that make up the cell walls and DNA structures of the cell nuclei are built. Free radicals and reactive oxygen species play in the polymorphs light eruption - from laymen often called sun allergy - a role. In sunscreen products so you can find a variety of substances to neutralize free radicals: Vitamin E, Vitamin C, rutin, Furalglucitol, 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 act with topical application as a free radical scavenger. [0038] A potent free radical scavenger is the flavonoid rutin, which is applied in combination with vitamin E as a pre-sun cream for the prevention of polymorphs Lichtdermatose days before the stay in the sun (Hadschiew 1997). The same principle is the recommendation, the skin at an early stage with a highly concentrated vitamin E cream (Example Optolind E) saturate (Henry 1994).

[0039] Almost every sunscreen contains vitamin E (tocopherol). Pure vitamin E creams achieve a sun protection factor of about 3. By oral intake can be in the epidermis do not achieve sufficiently high tocopherol concentrations. Moreover, studies show that can reduce the vitamin E concentration in the skin by sunlight by up to fifty percent (Thiele 1998). Therefore, a local application is required. When Vitamin E acetate penetrates well into the epidermis, where it is cleaved by esterases in free vitamin E. Due to its structure, it can be easily incorporated into the cell wall and protects them from attack by the radicals. [0040] To characterize the quality of a sunscreen to protect against sunburn in the future will no longer be in the foreground. While the SPF helps prevent erythema when used correctly, but it does not provide guidance on how the consumer can avoid immunosuppression or no longer repairable nuclear damage. This requires new measurement criteria. Acc Chem Res Vol Chemistry and Applications of Photocatalytic Oxidation of Thin Organic Films, 28, No: [0041] In the photocatalytic reaction among other things, produces very reactive free OH radicals, which have a strong antimicrobial activity (A. Heller..... . 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 is under photocatalytic activation by UV light a very strong reduction the output germ content in E. faecium to 0.01%, achieved in S. aureus on = 0.001% and 0.00002% to E. coli. This strong biocidal effect is undesirable in applications to the skin.

6.3. Anti-tumor effect

[0042] As the first indications of an effect of ingredients of Artemisia annua in various human cancers.

[0043] According to the results of the German Cancer Research Center in Heidelberg is also based this action on the formation of free radicals by the peroxide group in the presence of iron ions.

[0044] The test of artemisinin as a potential anti-cancer drug is still in its early stages.

[0045] triterpenoids isolated from Ganoderma concinna, apoptosis can initiate 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: 417-21). [0046] triterpenes, available applanatum from Ganoderma are against skin tumors of the mouse effective (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 trials (Murgo A, Cannon DJ, Blatner G, Cheson BD Clinical trials of MGI-114 Oncology 1999; 13:.. 233-8).

Proven [0047] has the supply of Lentinan in addition to chemotherapy in patients with gastric cancer, colorectal cancer and other tumors (Hazama S, Oka M, Yoshino S, Iizuka N, Wadamori K, Yamamoto et al. Clinical effects and immunological analysis of intraabdominaland intrapleural injection of lentinan for malignant ascites and pleural effusion of gastric carcinoma Cancer & chemotherapy 1995; 22:. 1595-7).

[0048] After the P 53 model (Nature 415, 26-27 (2002)), there are close links between low tumor morbidity and anti-aging effects.

6.4. Reduction of microbial contamination of implants

[0049] The infection rates in permanent implants are usually between 0.5 and 6% prevention strategies are urgently needed. Together with Candida staphylococci are the most common causative agent of a catheter-related sepsis, of which run approximately 50% fatal. In forming on the plastic surface organic film-the staphylococci are partially protected from the attack of antibiotics and develop very often a multiple resistance.

[0050] Each implant-associated infection is associated with a sharp increase in treatment costs associated [Can & Jansen, 2001]. , In Germany one goes out of 3000 to 4000 catheter-related deaths per year.

[0051] Antiseptic equipped implants can be used for example in the gastrointestinal and urogenital (Brewer et al., Biocompatibility, cell adhesion and degradation of surface-modified biodegradable polymer that designs for the upper unrinary tract. Tech. Urol. (1998), 4, 214-220;.. Multanen et al, Bacterial adherence to ofloxacin-displayed polyacetones-coated self-reinforced 1-lactide acid polymer uroological stent BJU International (2000), 900, 44-56; Schier wood, JM, H.-M. Wenchel, D. König, J. Beuth and G. Pulverer: importance of antimicrobial slow-release systems for preventing infections kathetherassoziierter Hyg Med 23 (1998) 548-556.....

[0052] The adhesion of microorganisms and cells to an implant is in the pathogenesis of nosocomial infections foreign body to plastic surfaces is the first important step. In PCT / EP 2008/056 730 from 05/31/08 method for coating surfaces with micro- and described nanoparticles by means of plasma processes, which can be used to prevent adhesion by germs. Working for coating medical devices with lipid nanoparticles produced from Artemisia annua are not available according to our research.

7. disadvantages of the prior art [0053] All combinations described in the prior art to use extracts of the whole plant Artemisia annua.

[0054] The use of the plant Artemisia annua focused so far on the use of artemisinin as an antimalarial agent. This requires the extraction with apolar solvents such as n-hexane. Here, the lipids are removed inevitably. In order for the conversion of biomass to micro and nanoparticles described in the prior art using the species-specific lipids is no longer possible.

[0055] The residual biomass and their ingredients remain largely ignored in previous valuation concepts.

The object of the present invention

[0056] A problem to be solved by the invention was to provide from the biomass of Artemisia annua means available that can be used after appropriate clinical testing for tumor treatment.

[0057] An additional objective was to provide lipid-containing micro- and nanoparticles for coating instruments and / or implants available.

[0058] One of primary object of the present invention is to find new applications for the remaining after separation of the Arteminsins from Artemisia annua residues and therefor provide new formulations.

Representation of the nature of the invention

[0059] The objects are achieved according to the features of the claims, on the one hand by preparations of biomass the plant Artemisia annua and the other by biomasses of lipid-containing algae or fungi, as well as the conversion of this preparation in the micro- and nanoparticles.

[0060] According to the invention novel particles having an average diameter which is depending on the make of between 10 nm and 10 microns is obtained, which integrate the ingredients of Artemisia annua and the respective combination partners. [0061] Depending on whether artemisinin-containing or Artemisin- free biomass are used from Artemisia annua, micro- and nanoparticles are obtained that can be used for very different tasks.

[0062] Micro- and nanoparticles, characterized in that are used for their production of biomass of the plant Artemisia annua, which contain all ingredients including artemisinin (sesquiterpene peroxides), on the one hand provide new means available that used after appropriate clinical testing for tumor treatment can be, on the other hand, means are also obtained which are suitable for the coating of surfaces and their antimicrobial properties.

[0063] In this application, the powerful free radical formation in the disintegration of the peroxide particularly in the presence of iron ions is used for cytotoxic and antimicrobial effects.

[0064] In order to protect themselves from the radical impact, including Artemisia annua radical scavenging agents. Micro- and nanoparticles, characterized in that are used for their production of biomass of the plant Artemisia annua, whose content of artemisinin (sesquiterpene peroxides) was removed prior to the conversion in the micro- and nanoparticles, use of these radical-scavenging properties. The micro- and nanoparticles produced under separation of peroxide structures are therefore suitable for anti-aging products, skin care and sunscreen products.

[0065] Regardless of whether artemisinin-containing or free Artemisin- biomasses are used, required for the achievement of the object conversion into micro- and nanoparticles with either the homogenization, the solvent can be effected Homogenisations- method or the solvent-emulsification process.

1. homogenization

[0066] The comminuted biomass of Artemisia annua and the residue after removal of the sesquiterpene peroxides are first heated. This fraction is suspended, dispersed or adsorbed in the fatty acid from other lipid-containing fungi or Cyanaobakterien) or the entire lipid-containing biomass. In parallel, a surfactant-water mixture is prepared. In this biomass one or more active substances (solid or liquid) can be added. 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. (Rotor stator principle) or by means of ultrasound is then prepared a pre-suspension with the aid of a stirrer. The pre-suspension is then homogenized using a high-pressure homogenizer, whereby the number of homogenization cycles and the working pressure according to the desired particle size and stability of the preparation be selected. Between cycles, ensure that the production temperature is set again and again. The surfactant serves to stabilize the suspension.

[0067] Should there be problems in the preparation of the level of temperature (eg. B. sensitive active ingredients) provide, so it is possible to perform the entire process, even at room temperature. In this case, the method in the same manner as described above, is performed, wherein the active substance to the lipid-containing organisms adsorbed or dispersed with addition of a small amount of water.

2. Solvent-Homogenisations- method

[0068] The comminuted biomass of Artemisia annua or extraction residues after removal of the sesquiterpene peroxides as well as lipid-containing biomass a second species and optionally selected active ingredients preferably Vitamin mixtures are suspended in a vaporizable organic solvent. Thereafter, this mixture is predispersed (stator-rotor principle or ultrasound), homogenized

(High-pressure homogenizer) and then spray dried or freeze-dried (Scheme 2). When freeze drying is to be noted that suitable cryoprotectants are used. Moreover, there is also the possibility of the organic solvent by suitable evaporator (z. B. rotary evaporator) to remove. The particles can be redispersed in a suitable aqueous surfactant solutions. Thereafter, a redispersion (stator-rotor principle or ultrasound) and homogenization (high pressure homogenizer) is necessary.

3. Solvent-emulsion method

[0069] This method is based on the preparation of an emulsion of water and a solution of Artemisia annua or the extraction residue of Artemisia annua after removal of the sesquiterpene peroxides in a suitable organic solvent. To an emulsifier is used to disperse the biomass drug. emulsifier and

Biomass are dissolved in a suitable organic solvent. To this solution, an aqueous phase containing a water-soluble co-surfactant is added and mixed with the lipid-containing biomass a second kind. Thereafter, this mixture is vordisp ergiert (stator-rotor principle or ultrasound). After a homogenisation step using a high pressure homogenizer, the organic solvent is removed by evaporation, wherein the active substance-containing biomass in the form of solid particles precipitated.

[0070] The biologically active ingredients from Artemisia annua, or with the combination partners can change according to charge and size distribution of the ingredients and the preparation conditions in the solid matrix (nano pellet) and / or in the lipid-containing envelope (nanocapsule), included in the envelope, and / or distributed in the dissolved or highly dispersed state in the formulation and / or adsorbed onto the edge zones of the nano-pellets or are adhered. By conversion of the combination of Artemisia annua and the lipid-containing biomass a second species in micro- and nanoparticles ingredients of both components are discharged controlled.

[0071] In the production of micro- and nanoparticles both artemisinin-containing and Artemisin- free biomasses with lipid-containing algae can be preferably combined with cyanobacteria with a lipid content of at least 10% and / or with the unicellular green alga Haematococcus pluvialis.

[0072] In a preferred embodiment of the invention, artemisinin-containing biomasses with microalgae are combined, which additionally have a natural content of Carbamidocyclophanen. Both Carbamidocyclophane containing biomasses of Cynanobakterien and the peroxide-containing structures of Artemisia annua exert a potent cytotoxic effect on tumor cells, but have different mechanisms of action, so that there is a synergistic effect. [0073] For this purpose particularly suitable are microalgae Carbamidocyclophane corresponding to the formula Image 1, characterized by a symmetrical carbon skeleton with Carbamidostrukturen in the side chain and by a different substitution pattern of the opposing the Carbamidostrukturen side chains, wherein Ri to R 7 are both halogen, hydroxyl, carbonyl - as may be alkyl groups also included. The Carbamidocyclophane of Formula 1 and preparations prepared therefrom image inhibit the growth of various tumor cell lines (MCF-7 breast cancer cells, 5637 bladder carcinoma cells) is particularly strong. Particularly preferred are micro- and nanoparticles that contain the active substance of formula 1 in a concentration of 0.01 to 3%.

[0074] The inventive transfer into micro- and nanoparticles occurs on the one hand to a synergistic cytotoxic effect of the combination of active substances of formula 1 and the image peroxide structures from Artemisia annua against

Tumor cells.

Figure imgf000016_0001
Formula 1: Carbamidocyclophane

[0075] In order to increase the cytotoxic potency, the proportion of the active substances of formula 1 or the image sesquiterpene peroxides can be increased in the inventive preparations by addition of the pure active ingredient.

[0076] When for the manufacture of micro- and nanoparticles, however artemisinin-free biomass and Carbamidocyclophan- used free microalgae are obtained particularly favorable skincare products. as lipid-containing biomass a second species are particularly suitable microalgae, which are characterized by a particular fatty acid composition in the lipid content for this task. By treatment with formulations containing micro- and nanoparticles produced according to the invention from Artemisia annua and microalgae of the skin's protective lipids are supplied. In addition, some microalgae have an extremely high water retention, which can be used in these formulations. Thus the trans-epidermal water loss is counteracted with the novel formulations. The inventive micro- and nanoparticles in skin moisture and grease is supplied in a particularly favorable form so that it appears smoother.

[0077] In a particularly advantageous embodiment is used as combination partners of Artemisia annua the unicellular green alga Haematococcus pluvialis. This green alga contains up to 5% of their biomass, the ketocarotenoid astaxanthin. The inventive microencapsulation Astaxanthin is protected and controlled released. [0078] Due to the radical scavenging properties of micro- and nanoparticles of the invention prevents the conditional by environmental factors such as UV light, chemical reagents, mechanical load, stress, heat and cold premature aging with regular use.

[0079] In another embodiment, both artemisinin-containing and Artemisin- free biomasses with lipid-containing fungi may be combined in the preparation of micro- and nanoparticles. Particularly preferred are the species Ganoderma lucidum and / or Ganoderma pfeifferi and / or Ganoderma applanatum and / or Ganoderma concinna and / or tsugae Ganoderma and / or Auricularia auricula-judae and / or Grifola frondosa and / or Hericium erinaceus and / or Lentinula edodes and / or Pleurotus ostreatus eryngii and / or Pleurotus.

[0080] If artemisinin-containing biomass or combinations of artemisinin-containing biomass and Carbamidocyclophan-containing microalgae with fungi of the genus Ganoderma, in particular combined, there is a synergistic effect.

[0081] Both Carbamidocyclophane of microalgae and the peroxide-containing structures of Artemisia annua exert a direct cytotoxic effect on tumor cells. [0082] In contrast, causes the biomass of the genus Ganoderma, induction of apoptosis and stimulation of an immune, in particular by the glucan content of Ganoderma species. Ingredients of Ganoderma species cause activation of phagocytosis by macrophages. The efficiency can be enhanced when combinations with triterpenes from Ganoderma concinna and / or Ganoderma applanatum and / or tsugae from Ganoderma lucidum, Ganoderma and / or inserted from Ganoderma pfeifferi. Advantageously, can be combined for this therapeutic purpose triterpenoids from these five Ganoderma species.

[0083] The interaction of cytotoxic effect, induction of apoptosis and macrophage activation is surprising and opens up new possibilities in the prevention and treatment of tumors. The micro- and nanoparticles according to the invention may contain cytostatic agents apoptosinduzierende agents and Immunstimmulantien than two or three combinations.

[0084] In contrast, if Artemisin- free biomass or combinations of Artemisin- free biomass and combined Carbamidocyclophan- free microalgae with mushrooms, which have proven themselves in promoting health and in the prevention, preparations with special care properties are obtained. The micro- and nanoparticles have strong free radical scavenger properties, which is of advantage in many applications. [0085] Also, due to the different fatty acid composition of fungi and / or cyanobacteria, the properties of the resulting products can be adapted to the requirements.

[0086] In particular Ganoderma pfeifferi suitable as combination partners for these applications. The metabolism of human cells (FL-cells) micro- and nanoparticles, made of G. pfeifferi is strongly stimulated. Surprisingly, it has been found that even if Ganoderma pfeifferi is greatly reduced in its concentration with micro- and nanoparticles made of a combination of Artemisia annua and Ganoderma pfeifferi similar strong stimulating effects are achieved in the cellular metabolism and the Artemisia annua component in large excess is present.

[0087] The ingredients of G. pfeifferi reduce oxidative stress and reinforce the endogenous radical defense. The pfeifferi verified only at G. Ganomycin ingredients A and B inhibit at a concentration of 10 ug / ml induced by the spontaneous luminol and oxygen radical release.

[0088] By controlling the votes ingredients proliferation is increased by human keratinocytes. Under the influence of the combination of glucose consumption increases from human cells, suggesting a stimulation of the metabolism. It was found that the respiration strengthened, encouraged the formation of proteins and cell aging is slowed. Under the same experimental conditions, no slowing the aging process can be detected in extracts from G. lucidum.

[0089] The increased under UV influence cell permeability - measured by determining the LDH activity in the medium - is reduced. The regenerative capacity of cells after UV damage is significantly increased. The micro- and nanoparticles also have the advantage of having strong radical scavenging properties, which is of advantage in many applications. the radical scavenging ingredients are released in a controlled both of Artemisia annua and G. pfeifferi from the micro- and nanoparticles.

[0090] It is not insignificant, moreover, that the production of special Ganoderma species that are not yet commercially exploited on a large scale, due to the slow fungal growth is very expensive. The combination partners from Artemisia annua against it is a by-product in the production of the drug against malaria. Therefore, with the inventive combination of Ganoderma species and Artemisia annua scrap an inexpensive way to produce an anti-aging agent with the cell metabolism stimulant properties is provided.

[0091] Particularly advantageous application properties are obtained when the remaining after removal of the sesquiterpene peroxides residues of Artemisia annua with biomasses are combined both from microalgae and from fungi species. Resulting from this three-way combination of micro- and nanoparticles are preferably suitable for use in sunscreen preparations.

[0092] This effect can be enhanced if vitamins are integrated into the micro- and nanoparticles. Vitamins may be added where they are integrated into the micro- and nanoparticles both in the homogenization, the homogenization solvent or solvent emulsion process during the production of micro- and nanoparticles. In a particularly preferred embodiment, vitamin E is used as acetate. The vitamins are from the micro and nano controls to the skin given. When Vitamin E acetate penetrates well into the epidermis, where it is cleaved by esterases in free vitamin E. Due to its structure, it can be easily incorporated into the cell wall and protects them from attack by the radicals. [0093] As play a special role in the development of skin tumors free radical damage is reduced with regular prophylactic use of sunscreens that contain the inventive micro- and nanoparticles, the risk of tumor development. Cells degenerate still under the influence of UV radiation are excited by the features contained in the preparations triterpenes from Ganoderma species, in particular from Ganoderma concinna to apoptosis.

[0094] In order to make the object, lipid-containing microparticles and nanoparticles for the coating of surfaces, for example of instruments and / or implants available has been solved by production of micro- and nanoparticles from which the biomass of Artemisia annua without separation of the peroxides. Here micro- and nanoparticles are formed with strong antimicrobial activity. These particles can be used to reduce the adhesion of proteins, cells and nuclei. In the unpublished at the filing date patent application PCT / EP2008 / 056730 of 31.05.2008, a method for coating surfaces is described procedure with micro- and nanoparticles with the aid of plasma that can be used to prevent adhesion by germs. The micro- and nanoparticles of the invention can be used for an extension of this process.

[0095] The features of the invention can be gathered from the claims also apparent from the description, and the individual features represent advantageous protectable embodiments, either singly or together in the form of combinations, is sought for with this document protection.

[0096] The invention will be explained with reference to embodiments, but is not limited to these examples.

embodiment

Example 1: composition of the fatty acid suitable for the production of micro- and nanoparticles of a combination of Artemisia annua with marine organisms microalgae

methodology

[0097] Pro-order were 5-6 strains examined each case with a duplicate.

Results

[0098] In Table 1, the variation in the levels found thereby indicated.

Figure imgf000020_0001

Figure imgf000021_0001

[0099] In the Chroococcales the tetradecanoic and tetradecenoic be found at a constant high level. Also strikingly high, the content of either n-hexadec-cis-9-enoic acid or 11-hexadecenoic. 8 more fatty acids each with a low content, typically <1% found.

[0100] In the Oscillatoriales the tetradecenoic acid was not found at all, the saturated C-14 acid was detected only in individual strains. n-hexadecanoic acid was mixed with a content of between 10 and 14% and n-octadeca-cis-9, cis 12, 15 ice-trienoic acid detected with a content between 20 and 30% for all strains. Also dominant was octadecadienoic acid, wherein the position of the double bond in the individual strains varied.

[0101] In the Nostocales dominates the n-hexadecanoic acid, which was demonstrated 26-72%, while the corresponding undersaturated acid was found only with a small proportion.

[0102] In contrast, in the majority at higher fatty acids with chain length polyunsaturated, forms, in particular the n-octadeca-cis-9, cis 12, 15 ice-trienoic acid or octadecatetraenoic acid. Since polyunsaturated fatty acids with large chain length of particular influence on the growth of S. aureus, this finding is particularly noteworthy. Further testing thereby the tribe has been found Bio 33 from the family of Nostocaceae be particularly suitable, whose fatty acid composition in Tab. 2 compared to other strains of this order is shown.

Table 2: Fatty acid composition in strains of the order Nostocales

Figure imgf000022_0001
[0103] There are combination partners from different orders with sufficiently high lipid content available. The properties of the microparticles and nanoparticles can be varied by selecting the appropriate for the particular application lipids.

Example 2 Antioxidant properties methodology

[0104] The determination of the antioxidant properties were thin-layer chromatography (CBS Camag Bibliography Service 88 SIEVERS A et al (2002) Simple thin-layer chromatography test for antioxidant compounds using DPPH assay.. 14-15).

Result

[0105] Figure 1 shows the qualitative DPPH test Artemsia annua / Ganoderma pfeifferi microparticles (left) compared to ascorbic acid (right).

[0106] As can be seen from the size of pfeifferi- Entfärbungszone the Artemsia annua / Ganoderma microparticles have powerful antioxidant properties of the studied microparticles.

Example 3 Influence of the microparticles of the invention on the cell metabolism

methodology

[0107] There was used a test arrangement (in DP 19709649.2 Jülich, W.-D., Woedtke, Th. By, Abel, P .: measuring arrangement for the detection of toxic, subtoxic, chronic toxic or stimulating effects of active ingredients and pollutants described using Perfusionszellkulturen). Confluent grown FL cells were treated to the inhibition of growth with mitomycin C.

Results

[0108] micro- and nanoparticles, made of G. pfeifferi, the metabolism of human cells stimulate strongly (FL-cells). [0109] With a combination of Artemisia annua and Ganoderma pfeifferi, erfϊndungsgemäß prepared by conversion into micro- and nanoparticles by using the species-specific lipids of G. pfeifferi could reach these effects, even if Ganoderma pfeifferi is greatly reduced in its concentration and the Artemisia αnnwα component in a large excess is present.

Example 4 Cytostatic effect Methodology

[0110] The Zytotxozität the Carbamidocyclophane and the carbamidocyclophanhaltigen combinations is carried out in vitro cultured MCF-7 cells, 5637 cells as well as to FL cells in 96-well microtiter plates. The MCF-7 cells and the 5637 cells 48 h in IMDM (Invitrogen) with 10% fetal bovine serum or the Fl-cells in Eagle's MEM + 8% fetal bovine serum pre-cultured (37 ° C, 5% CO2). Then, the Carbamidocyclophanhaltigen extracts of aquatic organisms or the isolated Carbamidocyclophane in concentrations of lOOμg / mL to 0.0000 be lμg / ml and the cells incubated on further 48h under the same conditions. The cells are then fixed and stained with crystal violet 0.02% to about 30 min. After washing and drying the cell-bound dye is dissolved with 70% ethanol and 550 nm ht the optical density at λ = Anthos II with a plate reader (Anthos, Salzburg, Austria) was measured (Bracht, K .; Boubakari; Grünert, R .; Bednarski, PJ, anti-Cancer drugs 2006, 17, 41-51) the inhibition of Zellpro life ration is calculated from the optical density measured in the batches incubated with the Carbamidocyclophanen or the carbamidocyclophanhaltigen extracts of aquatic organisms in comparison to the cell control (cells incubated with media only) and for the isolated Carbamidocyclophane the IC50 values ​​are calculated of values.

[Olli] result for the two tumor cell lines for the isolated Carbamidocyclophane IC50 values ​​between 0.7 ug / mL and 1.7μg / mL, for the Fl-cells, which are considered to be non-transformed cells, IC50 values ​​between 2.5 ug / mL, determined 3.1 ug / ml and 3.5 ug / mL. [0112] With micro- and nanoparticles, containing combinations of peroxides and sesquiterpene Carbamidocyclophanen, IC50 values ​​of less than 0.3 ug / mL are determined.

Example 5 Preparation of micro- and nanoparticles Table 3: formulation of the nano - and microparticles

Figure imgf000025_0001

[0113] The biomasses are combined and heated to a temperature of 80 0 C. An aqueous emulsifier solution is separated therefrom (80 0 C) heated to the appropriate temperature. Then both phases are combined and & using an Ultra Turrax T25 of the company. 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 reacted with a piston-gap high-pressure homogenizer Micron Lab 40 (APV-Gaulin, Lubeck) at a pressure of 500 bar and a temperature of 80 0 C homogenised four times.

Example 6 Determination of the minimum effective dose

Methodology of Example 3

6.1. Limiting the duration of application

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

Result:

Even after this short exposure time, a promoting effect reveals that at least 3 days detectable (Fig. 3). An extension of the exposure time to the 2% suspension to 72 hours increases the effect (Fig. 4), the enhancing effect is detectable over a longer period.

6.2. Limit the daily feeding to determine the minimum effective concentration, the daily intake of 0.5 ml of 2% strength suspension was limited, which was added within 111 min, ie, after which the cells were about 1329 min a medium without added active compound. The daily supply of the drug was reduced with this experimental setup to 0.012% of the cell culture medium.

Even at this low concentration, a promoting effect is seen (Fig. 5). Doubling the concentration (feed 2 times daily for 111 min), the effect more pronounced (Figure 6).

Claims

claims
1. micro- and nanoparticles of biomass the plant Artemisia annua one hand, and biomasses of lipid-containing algae or fungi on the other hand.
2. micro- and nanoparticles according to claim 1, characterized in that it is such a biomass of the plant Artemisia annua, containing all the ingredients including artemisinin (sesquiterpene peroxides).
3. Micro- and nanoparticles according to claim 1, characterized in that it is such a biomass of the plant Artemisia annua, whose content of artemisinin (sesquiterpene peroxides) was removed prior to the conversion in the micro- and nanoparticles.
4. micro- and nanoparticles according to claim 1 to 3, characterized in that it is in used for the preparation of lipid-containing algae, cyanobacteria with a lipid content of at least 10% and / or cyanobacteria with a lipid content of at least 10% and / or the unicellular green alga Haematococcus pluvialis is.
5. Micro- and nanoparticles according to any of claims 1 to 4, characterized in that it is in the selected lipid-containing cyanobacteria strains with a natural content of Carbamidocyclophanen
6. micro- and nanoparticles according to any one of claims 1 to 4, characterized in that the Carbamidocyclophan-containing strains selected compounds of formula 1
Figure imgf000027_0001
Preferably in a concentration of 0.01% to 3%, where R can stand to R 7 are both halogen, hydroxyl, carbonyl and alkyl groups, or those compounds of formula 1 the micro- and nanoparticles in the manufacture are added until a concentration of 0.01 to 3% was reached.
7. micro- and nanoparticles, characterized in that active substances of formula 1 and image sesquiterpene peroxides are from Artemisia annua combined.
8. micro- and nanoparticles according to claim 1 to 7, characterized in that it is in the used for producing lipid-containing mushrooms to the mushroom species
• Ganoderma lucidum and / or
• Ganoderma pfeifferi and / or
• Ganoderma applanatum and / or
• Ganoderma concinna and / or
tsugae • Ganoderma and / or
• Auricularia auricula-judae and / or
• Grifola frondosa and / or
• erinaceus Hericium and / or
• Lentinula edodes and / or
• Pleurotus ostreatus and / or
• Pleurotus eryngii is.
9. micro- and nanoparticles, characterized in that active substances of formula 1 and image biomass combined from Ganoderma species.
10. micro- and nanoparticles, characterized in that sesquiterpene peroxides from Artemisia annua and biomass combined from Ganoderma species.
11. micro- and nanoparticles, characterized in that active substances of formula 1 image, sesquiterpene peroxides from Artemisia annua and biomass combined from Ganoderma species.
12. micro- and nanoparticles according to any of claims 1 to 11, characterized in that it additionally mindestes one of vitamins A, C or E contained, wherein vitamin E is preferably used as the acetate.
13. A process for the preparation of micro- and nanoparticles according to any one of claims 1 to 12, characterized in that either comminuted biomass from Artemisia annua, which contains all active ingredients or, alternatively, biomass from Artemisia annua which the content of artemisinin (sesquiterpene peroxides) was removed, mixed with the algae or fungi according to claim 1 and / or additives and is converted by known processes into microparticles and nanoparticles.
14. Use of the micro- and nanoparticles according to any of claims 1 to 13 for cosmetic and / or pharmaceutical preparations and / or as dietary supplements.
15. Use of compositions according to claim 3 as a) means for slowing down the cellular aging and / or b) sunscreens and / or c) means with anti-aging effect. d) means for the prophylaxis of skin tumors e) means for inducing apoptosis in tumor cells
16. Use of compositions according to one of claims 2, 9, 10, 11, 13 as a) means with anti-tumor action b) means for treating tumors c) means for inducing apoptosis in tumor cells
17. The use of micro- and nanoparticles according to any of claims 1 to 3 for the coating of surfaces.
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US20070269537A1 (en) * 2003-02-10 2007-11-22 Bioderm Research Skin Condition Improvement Including Acne, Rosacea, and Topical Wounds by Artemisia Annua Extract via Iron Siderophore Trojan Horse Delivery System
WO2004075907A2 (en) * 2003-02-27 2004-09-10 Ernst-Moritz-Arndt-Universität Greifswald Salutary compositions consisting of fungi containing lipids and thiocyanates
JP2006143711A (en) * 2004-10-22 2006-06-08 Taisho Pharmaceut Co Ltd Oral composition for ameliorating aging note

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