WO2001079245A1

WO2001079245A1 - Novel flavone glycoside derivatives for use in cosmetics, pharmaceuticals and nutrition

Info

Publication number: WO2001079245A1
Application number: PCT/EP2001/004151
Authority: WO
Inventors: Ralf Otto; Bernadette Geers; Albrecht Weiss; Dirk Petersohn; Kordula Schlotmann; Klaus Rudolf SCHRÖDER
Original assignee: Henkel Kommanditgesellschaft Auf Aktien; Cognis Deutschland Gmbh & Co. Kg
Priority date: 2000-04-18
Filing date: 2001-04-11
Publication date: 2001-10-25
Also published as: EP1274712A1; JP2004501086A; AU2001250423A1; DE10019235A1

Abstract

The invention relates to flavone and isoflavone glycoside derivatives of general formula (I): [A1-C(=O)O]m-[X-O-Z]-[O-C(=O)-A2]n (I), wherein [X-O-Z] represents a flavone or isoflavone glycoside structure, wherein X represents a flavone or isoflavone parent substance of formula (IIa) or (IIb), said (iso)flavone parent substance being mono- or multisubstituted and/or mono- or multireduced (hydrogenated), wherein Z (sugar) represents a mono-, di- or polysaccharide which is acetally bonded to the radical X and is ester-substituted with A2 n-times, [A1-C(=O)] representing an acyl radical on the flavone or isoflavone parent substance, wherein A1 and A2, independently of each other, represent a polyunsaturated C15- C25- alkenyl radical with at least 4 isolated and/or at least 2 conjugated double bonds or an arylaliphatic radical with 1-4 methylene groups between the ester group and the aromatic ring, wherein [C(=O)A2] represents an acyl radical on the sugar Z, wherein n is a whole number (1, 2, 3, ...) but not 0, wherein m is a whole number including 0 (0, 1, 2, 3, ...) and wherein R1, R2 and R3 represent hydroxyl groups or hydrogen atoms.

Description

New flavone glycoside derivatives for use in cosmetics, pharmaceuticals and nutrition

The present invention relates to new, biologically active flavone and isoflavone glycoside derivatives of the general formula (I)

[A ₁ -C (= 0) O] _m - [XOZ] - [OC (= O) -A ₂ ] _n (I) of aliphatic and arylaliphatic carboxylic acids, process for their preparation and cosmetic and / or pharmaceutical containing them Preparations, as well as their use as additives in food and feed.

The use of active ingredients is becoming increasingly important in cosmetics. The active ingredients that are already used in cosmetics are not always natural substances. The optimization of known active ingredients and the production of new active ingredients are the subject of much research.

In the broadest sense, active substances are substances that - occurring or supplied in relatively small amounts - can have a great physiological effect. Here one has to think of hormones, vitamins, enzymes, trace elements etc., but also of pharmaceuticals (medicinal substances), feed additives, fertilizers and

Pesticides. It is not uncommon to see synergism.

Flavones and isoflavones / flavonoids and isoflavonoids or flavone fluosides and isoflavone qlycosides

Flavones are 2-phenyl-4H-1-benzopyran-4-ones, in which hydroxyl groups are present or may be absent at different positions on the rings. An example of a flavone is apigenin, the chemical name of which is 2- (p-hydroxyphenyl) -4H-1- (5,7-dihydroxybenzopyran-4-one) (see Römpp Chemie-Lexikon, 9th edition, volume 2, p. 1373/4) The additional hydroxyl groups are located on the phenyl and / or the benzopyran ring, as the example shows, in other words: for the purposes of the present invention, flavones are understood to mean substances which contain hydrogenation, Represent oxidation or substitution products of 2-phenyl-4H-1-benzopyran-4-one, where hydrogenation can take place in the 2,3-position of the carbon skeleton, and with substitution the replacement of one or more hydrogen atoms by hydroxy or methoxy Groups to be understood. This definition therefore includes flavans, flavan-3-oles (catechins), flavan-3,4-diols (leucoanthocyanidins), flavones, flavonols and flavanones in the conventional sense. In addition to apigenin, the flavones according to the invention include, for example, chrysine, galangin, fisetin, luteolin, camphor oil, quercetin, poppy seeds, robinetin, gossypetin, taxifolin, myricetin, rhamnetin, isorhamnetin, naringenin, eriodyctiol, hesperetin, liquiritigenin, catechin and epicatechin.

For the purposes of the present invention, however, isoflavones are understood to mean substances which are hydrogenation, oxidation or substitution products of 3-phenyl-4H-1-benzopyran-4-one, with hydrogenation in the 2,3-position of the carbon skeleton can take place, and wherein substitution is to be understood to mean the replacement of one or more hydrogen atoms by hydroxyl or methoxy groups. The isoflavones according to the invention include, for example, daidzein, genistein, prunetin, biochanin, Orobol, Santal, pratensein, irigenin, glycitein, biochanin A and formononetin.

Flavones and fiavone glycosides (flavonoids) such as aspartin, orientin (lutexin), cisorientin (lutonaretin), isoquercitin, rutin, naringin and the above-mentioned, but also isoflavones and isoflavone glycosides (isoflavonoids) are known to be scavengers of oxygen radicals and the skin, thereby causing protease radicals and inhibitors they can actively counteract aging of the skin and scarring. Because of their coloring properties, some flavones such as quercetin are used as food colors. At the same time, due to their ability to capture oxygen radicals, they also act as antioxidants. Some flavonoids are inhibitors of aldose reductase. This plays a crucial role in the development of diabetes damage (vascular damage, cataracts). Other flavonoids (such as hesperidin and rutin) find therapeutic use particularly as vasodilator, capillary active agents. The derivatizations carried out according to the invention achieve an improved effect and an increased bioavailability, as has already been shown using salicine derivatives as an example.

Many naturally occurring alkyl and phenol glucosides show antiviral, antimicrobial and sometimes anti-inflammatory effects. However, due to their polarity, they are often not very bioavailable or their selectivity is too low. For example, salicin (a glycosidic active ingredient from willow bark) is a non-steroidal anti-inflammatory agent (NSAIA) that shows significantly improved effectiveness after derivatization (esterification). Recently, new arylaliphatic salicine esters such as phenylacetoyl-salicin or phenylbutyroyl-salicin were successfully synthesized, the esterification preferably taking place on the primary OH groups of salicin (first on the sugar, then on the benzyl radical) in the salicin. Due to the arylaliphatic residue, the mass transfer to the site of action is improved and the selectivity of the action is increased. In contrast to unmodified salicin, these derivatives preferentially inhibit prostaglandin synthase 2 (lower risk of side effects) (Ralf T. Otto, biotechnological production and characterization of new pharmaceutically active glycolipids, dissertation (1999) ISBN 3-86186-258-1).

PUFAs and CLAs

The PUFAs (polyunsaturated fatty acids) and CLAs (conjugated linoleic acids) belong to the group of essential fatty acids in the diet and also show a positive effect when used in the prophylaxis of arteriosclerosis. In addition, pharmaceutical effects are also important: they can have anti-inflammatory effects (inhibition of prostaglandin or leukotriene synthesis), but also thrombolytic and hypotensive effects.

According to the invention, PUFA is defined as a polyunsaturated fatty acid with 16 to 26 carbon atoms, the fatty acid having at least four isolated and / or at least two conjugated double bonds. Examples of PUFAs are the twelve octadecadienoic acids isomeric to linoleic acid (ice, ice, 9,12-octadecadienoic acid) (which occur in nature), which have conjugated double bonds at the carbon atoms 9 and 11, 10 and 12, or 11 and 13.

These isomers of linoleic acid (e.g. ice, trans, 9,11-octadecadienoic acid, trans, ice, 10,12-octa-decadienoic acid, ice, ice, 9,11-octadecadienoic acid, trans, ice, 9,11-octadecadienoic acid, trans, trans, 9,11-octadecadienoic acid, ice, ice, 10,12-octadecadienoic acid, ice, trans, 10,12-octadecadienoic acid, trans, trans, 10,12-octadecadienoic acid) can be prepared in a conventional manner by chemical isomerization of linoleic acid, depending on the reaction conditions, these reactions only lead to CLA mixtures of various compositions (eg Edenor UKD 6010, Henkel KGaA). Because of their conjugated double bonds, these isomeric octadecadienoic acids are also referred to as "conjugated linoleic acids" (CLAs).

Although numerous pharmacologically active substances have already been described in the literature, which intervene, for example, in the inflammatory cascade, there is still a need for more effective active substances which are low in side effects. There is also a need for active ingredients with good resorbability and rapid penetration into the skin, which must also be able to be incorporated well into pharmaceutical or cosmetic formulations.

There is also a particular interest in the discovery of active ingredients that can prevent the aging processes of human skin.

Human skin is the largest organ in the human body. It is a very complex organ, which consists of a variety of different cell types and forms the body's interface with the environment. This fact makes it clear that the cells of the skin are particularly exposed to exogenous signals of the environment, physical and chemical in nature. Many of these exogenous pollutants contribute to the aging process of the skin.

The macroscopic phenomena of aging skin are based on the one hand on intrinsic or chronological aging, and on the other hand on extrinsic aging due to environmental stress. The ability of living skin cells to respond to your environment react changes with time. There are aging processes that lead to senescence and ultimately cell death. The visible signs of aged skin are to be understood as an integral of intrinsic and extrinsic aging (eg due to sunlight), the events of extrinsic aging accumulating in the skin over a longer period of time.

Exogenous signals are received by cells and lead to changes in the gene expression pattern, partly via complex signal transduction cascades. In this way, each cell responds to signals from its environment by adapting its metabolism. For example, the cells of the skin notice the high-energy radiation from the sun and react to this by changing their RNA and protein synthesis capabilities. Some molecules are increasingly synthesized after a stress stimulus (e.g. sunlight) (e.g. the collagenase MMP-1), others are produced to a lesser extent (e.g. collagen). Furthermore, there will be no significant change in a large number of the synthesis processes (e.g. TIMP-1).

Induction of collagenase MMP-1 by sunlight or other stress factors is considered to be the main cause of the process of extrinsic skin aging. The collagenase MMP-1 destroys the most important component of the connective tissue of the skin, the collagen, and among other things leads to a reduction in skin elasticity and the formation of deep wrinkles. In young and not stressed skin, the activity of collagenase is regulated by a naturally occurring inhibitor TIMP-1 (Tissue Inhibitor of Matrix Metalloprotease-1). There is an extremely sensitive balance between MMP-1 and TIMP-1, which is disturbed by exogenous stress. The expression of MMP-1 is affected by skin stress, e.g. increased exposure to sunlight. In contrast, the synthesis of the inhibitor TIMP-1 is not changed significantly. Therefore it comes under the influence of exogenous stress, e.g. Sunlight, excessive collagen breakdown in the skin. The result is premature aging of the skin.

Efforts to cosmetically treat the effects of stress-induced skin aging are aimed at reducing MMP-1 activity or increasing the synthesis of collagen. By using retinoic acid or retinol, the MMP-1 Synthesis in the skin is reduced or collagen synthesis is increased. However, the use of retinoic acid for cosmetics is not permitted in Europe due to its teratogenic properties. Cytotoxic effects, insufficient stability in formulations, undesirable side effects or even the disturbing intrinsic color lead to the limitation of the cosmetic use of active ingredients, such as alpha-tocopherol, propyl gallate or various plant extracts.

The object on which the present invention is based was therefore to provide substances which have few side effects, have a good action and are easy to process and apply.

Flavone and isoflavone glycosides are e.g. known from nature. On the other hand, esters of such flavone and isoflavone glycosides in which at least one of the hydroxyl groups of the sugar is esterified with an (unsaturated) carboxylic or fatty acid, and in which possibly still are not known (neither from plants, microorganisms or animal cells nor synthetically produced) there is another ester grouping between one of the hydroxyl groups of the flavone or isoflavone portion and another unsaturated fatty acid.

Surprisingly, the inventors found that certain flavone and isoflavone glycoside esters have improved bioavailability, enhanced activity and / or a broader spectrum of activity than the known individual components (fatty acid or (iso) flavone glycoside). In these (iso) flavone glycoside derivatives, the flavones or isoflavones are glycosidically linked to at least one sugar via at least one hydroxyl group. The sugar can be linked to the (iso) flavone residue via an OH group on the benzopyran ring or via an OH group on the phenyl ring. The group [Aι-C (= O)] can also be linked to the (iso) flavone residue via an OH group on the benzopyran ring or via an OH group on the phenyl ring. It is preferred if the sugar is linked via the benzopyran ring and the fatty / carboxylic acid also via the benzopyran ring or via the phenyl ring of the (iso) flavone radical. Suitable sugars are mono- and oligosaccharides, in particular D-glucose, D-galactose, D-xylose, D-apiose, L-rhamnose, L-arabinose and rutinose. Examples of the flavone glycoside residues in the compounds according to the invention are rutin, hesperidin and naringin. Preferred examples of the isoflavone glycoside residues in the compounds according to the invention are daidzin and genistin.

By providing the compounds of the present invention, the inventors were able to achieve the object.

The compounds of the present invention are flavone and isoflavone glycoside

Derivatives of the general formula (I):

[A ₁ -C (= O) O] _m - [XOZ] - [OC (= O) -A ₂ ] _n (I), where [XOZ] is a flavone or isoflavone glycoside structure, where X is a flavone or isoflavone base body of the formula (Ila) or (Ilb)

(Ila) (Ilb)

represents, wherein the (iso-) flavone basic body is mono- or polysubstituted and / or mono- or polysubstituted (hydrogenated), where Z (sugar) stands for a mono-, disaccharide or polysaccharide, which is acetally bound to the radical X and n- is substituted ester-like with A ₂ . wherein [Aι-C (= O)] represents an acyl radical on the flavone or isoflavone base body, in which A ^ and A ₂ independently of one another are a polyunsaturated C ₁₅ - C2 ₅ -

Alkenyl radical with at least 4 isolated and / or at least two conjugated double bonds or an arylaliphatic radical with 1-4 methylene groups between

Represent ester group and aromatic ring, in which [C (= O) A ₂ ] represents an acyl radical on the sugar Z, in which n is an integer (1, 2, 3, ...) but not 0, in which m is an integer (1, 2, 3 , ...) including 0, and wherein R1, R2, R3 represent hydroxyl groups or hydrogen atoms.

Preferred sugar fractions are generally those Z which mean a monosaccharide. The following monosaccharides are particularly preferred: rhamnose, threose, erythrose, arabinose, lyxose, ribose, xylose, allose, altrose, galactose, glucose, gulose, idose, mannose, talose and fructose, the naturally occurring stereoisomers of sugar being the preferred form in each case , Also preferred are disaccharides which are built up from the monosaccharides mentioned above, the naturally occurring stereoisomers of sugar being the preferred form in each case.

According to a preferred embodiment, the binding of the (iso) flavone body to the sugar takes place via a primary alcohol group of the sugar (e.g. via OH to Cβ of the glucose). According to a further preferred embodiment, Z-O-X is the naringin skeleton with the formula (III)

(III)

Further preferred flavones / flavonoids (X or X-O-Z) in the general formula (I) are asparagine, orientin (lutexin), cisorientin (lutonaretin), isoquercitin, naringin, rutin, camphor oil and quercetin.

Preferred compounds of the general formula (I) are especially those in which XOZ is naringin according to formula (III) and A _{2 are} the acyl radicals of the following acids: p- Chlorophenylacetic, hydrocinnamic, stearic, 12-hydroxystearic, palmitic, lauric, oleic, coumaric, capric, cinnamon, 4-phenylbutter, 4-hydroxyphenylacetic, 5-phenylvaleric acid or those under the trade names available mixtures Edenor UKD 6010 and UKD 7505. Edenor UKD 6010 and UKD 7505, p-chlorophenylacetic and hydrocinnamic acid are particularly preferred acids. It is particularly preferred for all of these combinations of naringin and the fatty acids mentioned if n = 1 or n = 2 and m is 0 at the same time. If n = 1 (and m = 0), the preferred position of A _{2 is} the primary OH group on the sugar portion in formula (III). However, all secondary OH groups of the sugar can also be considered as preferred embodiments for the esterification. If n = 2 (and m = 0), it is preferred if one esterification occurs on the primary OH group and the second on one of the secondary OH groups of the sugar. Especially on one of the two secondary OH groups on the same or at one of the three secondary OH groups of the second six-membered ring.

Further preferred compounds of the general formula (I) are those in which X-OZ is naringin, A _{2 are} the acyl residues of the following acids: p-chlorophenylacetic acid, hydrocinnamon, stearic, 12-hydroxystearic, palmitic, lauric, oleic, coumaric, capric, cinnamon, 4-phenylbutyric, 4-hydroxyphenylacetic, 5-phenylvaleric acid or the mixtures available under the trade names Edenor UKD 6010 and UKD 7505; n = 1 or n = 2 and m is 1 at the same time. When n and m are both 1, the preferred position of A _2, the primary OH group in the sugar ₌ Ai either the 5-OH group of the benzopyran or the 4'-hydroxy group of the phenyl ring. But as in the case when m = 0, A _{2 can} also be esterified via all secondary OH groups of the sugar. If n = 2 and m is 1 at the same time, it is preferred if the one esterification of A ₂ on the primary OH group and the second on one of the secondary OH groups of the sugar, in particular on one of the two secondary OH groups on same or on one of the three secondary OH groups of the second six-membered ring, and the esterification of A ^ takes place via the benzopyran or the phenyl ring.

The inventors have found that the compounds of the general formula (I) can surprisingly be obtained by mild lipase-catalyzed esterifications. Accordingly, the present invention further provides a process for the preparation of the compounds of the formula (I) according to the invention. The process according to the invention is characterized in that an acetal (from sugar and flavone / isoflavone base body) with a polyunsaturated fatty acid (with at least four isolated double bonds or with at least two conjugated double bonds) such as a conjugated linoleic acid (octadecadienoic acid), with an arylaliphatic carboxylic acid , is esterified or transesterified with an ester of these carboxylic acids or with an activated fatty acid derivative under the catalytic action of one or more enzymes. Esterification at primary OH groups of the sugar is preferred, but secondary alcohol groups of the sugar can also be esterified.

Suitable enzymatic catalysts for the esterification of the acids mentioned and the acetal components containing hydroxyl groups include the hydrolases, especially the lipases (ester hydrolases) such as the lipases from Candida rugosa (formerly Candida cylindracea), Candida antarctica, Geotrichum candidum, Aspergillus niger, Penicillium roqueforti, Rhizopus arrhizus and Mucor miehei.

A preferred lipase is the lipase (isoenzyme B) from Candida antarctica, for which there are two reasons. First, it shows a particularly high selectivity in the esterification of the acetals with the unsaturated fatty acids, although these are not among their typical substrates. Furthermore, it shows no interface activation (a crucial feature for the classification of hydrolases in the group of lipases), since it lacks an important lipase structural feature, a movable peptide chain at the active center (so-called lid).

In the preparation of the compounds according to the invention by the customary methods of chemical synthesis, because of the presence of several free hydroxyl groups of the sugar and / or flavone / isofiavone base body, mixtures of single and multiple esterified products are generally formed, so that the introduction and removal of protective groups is necessary if you want to specifically synthesize a particular compound. Targeted esterification, however, is crucial for the biological availability and compatibility of the substances according to the invention. However, chemical synthesis leads to coarse product mixtures due to a lack of regioselectivity. Therefore, the enzymatic (see examples) mild and regioselective synthesis described here is advantageous. According to the invention, region-specific means that only a certain OH group of a polyol is esterified. Correspondingly, regioselective means that a particular OH group of a polyol is preferably, but not exclusively, esterified.

Once the compounds of the formula (I) according to the invention have been prepared using the process according to the invention, a process must generally follow in order to purify the desired compound (s). It is therefore a further object of the present invention to provide a process for the purification of the compounds of the formula (I), which is characterized in that it is an aqueous two-phase extraction process with organic solvents, by means of which the target compound is selective from the non implemented fatty acids can be separated. The organic solvent is preferably n-hexane, cyclohexane, THF, dieethyl ether. Alternatively, the purification can also be carried out by a chromatographic process on silica gel, preferably with ethyl acetate / methanol or dichloromethane / methanol mixtures with small proportions of acetic acid and / or water, which can also be carried out in addition to an aqueous two-phase extraction process with organic solvents.

Since the flavone / isoflavone glycosides of the formula (I) have good bioavailability and activity, they can be used in cosmetic and pharmaceutical preparations and / or as food additives, with the result that the quality of these products is significantly improved.

The compounds of the formula (I) according to the invention have an inhibition of skin proteases (anti-aging, anti-wrinkling), an antioxidative potential, skin-lightening effect and an inhibition of transcription. That is particularly surprising skin-lightening effect (as a result of an inhibition of tyrosinase) of these compounds, in particular the good skin-lightening effect of the compounds according to the invention in which ZOX means naringin and the primary OH group of which is esterified either with phenylpropionic acid, with hydroxyphenylacetic acid or with p-chlorophenylacetic acid.

It has furthermore been found that the compounds of the formula (I) according to the invention, in particular those compounds in which ZOX is naringin and the primary OH group of which is esterified either with phenylpropionic acid, with hydroxyphenylacetic acid or with p-chlorophenylacetic acid, are capable of causing sunlight To influence expression of MMP-1, TIMP and Colαi in a cosmetically desired manner and thus to counteract the loss of collagen in the Dennis. As a result, these compounds are outstandingly suitable for a cosmetic treatment of the skin, by means of which aging of the skin induced by sunlight is prevented and / or by which the consequences thereof are reduced.

The formation of collagen is influenced in particular by the extent of the expression of MMP and TIMP. The following strategies are possible for analyzing the factors involved in the process of homeostasis of sunlight-exposed skin cells: a) MMP: - quantification of the enzyme activity of MMP-1

- Quantification of the synthesized MMP-1 protein.

- Quantification of the synthesized MMP-1 mRNA .. b) TIMP - Quantification of the synthesized TIMP protein.

- quantification of the synthesized TIMP mRNA .. c) collagen - quantification of the synthesized collagen protein.

- Quantification of the synthesized Colαi mRNA ..

The production of mRNA is the first and most important step in the course of protein synthesis. Active substances that have an effect on mRNA production therefore automatically also have an effect on the amount of protein and enzyme activity. In a subsequent step, the effects of the effects on mRNA production can be demonstrated by the detection of the protein collagen in the skin model itself. In the context of the invention it was possible to show that naringin derivatives according to the invention are able to reduce the expression of MMP, to increase the expression of TIMP, to increase the expression of COICH and to increase the formation of collagen.

In photo-aged skin, MMP-1 is predominantly produced by fibroblasts, but the reactions of the skin to stress should not be viewed as reactions of individual, isolated skin cells. Rather, every cell is integrated into a complex communication network. This network effects the exchange of information directly between neighboring cells, but also between cells located further apart, e.g. the cells of the epidermis and dermis. The communication mechanisms between the cells of the skin include signaling molecules such as Interleukins, growth factors (e.g. KGF, EGF or FGF) etc. involved. For this reason, the analysis of the drug effects was carried out on skin models consisting of a dermal and an epidermal compartment.

It has furthermore been found that the compounds according to the invention are considerably less phototoxic compared to conventional active substances against photoaging of the skin.

In addition, the compounds according to the invention can be incorporated particularly well into lipophilic base formulations and can be easily formulated as stable emulsions.

Accordingly, the compounds of formula (I) according to the invention are used for the production of cosmetic and / or pharmaceutical preparations and / or food or feed. The compounds according to the invention can be present or used in the form of the pure substance or as a mixture of plant extracts of different origins. The (iso-) flavones and their glycosides are preferably used in the preparations / additives as components of a mixture of substances obtained from a plant, in particular a vegetable extract. Such vegetable substance mixtures can be obtained in a manner familiar to the person skilled in the art, for example by pressing or extracting them from plants such as citrus plants (family of the Rutaceae) or acacias.

The invention further relates to the use of the compounds of the formula (I) for the production of cosmetic and / or pharmaceutical preparations; the use as food supplements or additives in food preparations and in feed (e.g. for animal breeding); cosmetic and pharmaceutical preparations as well as food preparations and animal feeds containing (a) compound (s) of the formula (I).

The cosmetic preparations obtainable using the compounds (I) according to the invention, such as hair shampoos, hair lotions, bubble baths, shower baths, creams, gels, lotions, alcoholic and aqueous / alcoholic solutions, emulsions, wax / fat compositions, stick preparations, powders or ointments as auxiliaries and additives, mild surfactants, oil bodies, emulsifiers, superfatting agents, pearlescent waxes, consistency agents, thickening agents, polymers, silicone compounds, fats, waxes, stabilizers, biogenic agents, deodorants, antidandruff agents, film formers, swelling agents, UV light protection factors, antioxidants, hydrotropes, Contain preservatives, insect repellents, self-tanners, solubilizers, perfume oils, dyes, germ-inhibiting agents and the like.

The amount of the compounds according to the invention in the cosmetic (but also pharmaceutical) preparations is usually in the range from 0.01 to 5% by weight, but preferably in the range from 0.1 to 1% by weight, based on the total mass of the preparations ,

To prepare pharmaceutical or cosmetic preparations, the compounds of the general formula (I) according to the invention, optionally in combination with other active substances, together with one or more inert substances usual carriers and / or diluents, e.g. B. with corn starch, milk sugar, cane sugar, microcrystalline cellulose, magnesium stearate, polyvinylpyrrolidone, citric acid, tartaric acid, water, water / ethanol, water / glycerin, water / sorbitol, water / polyethylene glycol, propylene glycol, carboxymethyl cellulose or fatty substances such as hard fat or their suitable mixtures , in common galenical preparations such as tablets, dragees, capsules, powders, suspensions, drops, ampoules, juices or suppositories.

The daily dosage required to achieve a corresponding effect in pharmaceutical applications is advantageously 0.1 to 10 mg / kg body weight, preferably 0.5 to 2 mg / kg body weight.

The food substitutes and additives obtainable using the compounds of the formula (I) according to the invention, such as athletic drinks, suitably contain the compound (s) of the formula (I) in an amount which, with a customary need for fluid intake of 1 to 5 liters per Day leads to a dosage of these compounds of 0.1 to 10 mg, preferably 0.5 to 5 mg, per kg of body weight. An example of use in the food industry is for the compounds of the formula (I) as colorings and / or spices.

Examples

Example 1: Preparation of 6-O-cis-9, trans-11-octadecadienoyl-naringin

2 g D - (-) - naringin 5 g, CLA (Edenor UKD 6010), 12 g molecular sieve, 15 ml t-butanol and 10 g immobilized lipase B from Candida antarctica were stirred for 40 hours at 60 ° C and 100 rpm in a magnetic stirrer Incubated 250 ml Erlenmeyer flasks. The reaction was carried out by means of thin layer chromatography (silica gel KG60 plates with fluorescent indicator; eluent: ethyl acetate / methanol 10: 1 v / v; visualization: UV detection and by means of acetic acid / sulfuric acid / anisaldehyde (100: 2: 1 v / v / v) immersion reagent The product was extracted with 20 ml of n-hexane and purified by column chromatography (silica gel F60; mobile phase: ethyl acetate / methanol 10/1 v / v). Rf value: 0.47 (ethyl acetate / methanol 10: 1)

Example 2: Preparation of 6-O-naringin (3-phenyl-propionic acid) ester

5.8 g of naringin, 1.5 g of 3-phenylpropionic acid, 3.7 g of molecular sieve, 15 ml of t-butanol and 11 g of immobilized lipase B from Candida antartica were incubated for 24 hours at 60 ° C. and 100 rpm in a 250 ml flask , The reaction was followed by thin layer chromatography (silica gel 60 F ₂₅₄ ; eluent ethyl acetate / methanol 10: 1 v / v; visualization by UV detection). When the reaction was stopped, the conversion based on naringin was 20%. The product was extracted with 20 ml of n-hexane and purified by column chromatography (silica gel F60; eluent: ethyl acetate / methanol 10/1 v / v).

Rr value: 0.16 (ethyl acetate / methanol 10: 1 v / v) yield: 0.85 g

The column chromatographic separation was not optimized. In addition to fractions with the pure product, mixed fractions were also obtained which contained unreacted naringin. To determine the stated yield, only those fractions from column chromatography which only contained the desired product were used. Example 3: Preparation of 6-O-naringin (p-CI-phenylacetic acid) ester

5.8 g of naringin, 1.7 g of p-chlorophenylacetic acid, 3.8 g of molecular sieve, 15 ml of t-butanol and 11 g of immobilized lipase B from Candida antartica were incubated at 60 ° C. and 100 rpm in a 250 ml flask for 24 hours , The reaction was followed by thin layer chromatography (silica gel 60 F ₂₅₄ ; eluent ethyl acetate / methanol 10: 1 v / v; visualization by UV detection). When the reaction was stopped, the conversion based on naringin was 20%. The product was extracted with 20 ml of n-hexane and purified by column chromatography (silica gel F60; eluent: ethyl acetate / methanol 10/1 v / v).

Rr value: 0.20 (ethyl acetate / methanol 10: 1 v / v) yield: 0.50 g

The column chromatographic separation was not optimized. In addition to fractions with the pure product, mixed fractions were also obtained which contained unreacted naringin. To determine the stated yield, only those fractions from column chromatography which only contained the desired product were used.

Example 4: Production of further naringin derivatives

Naringin derivatives, prepared as described in Example 1 (reaction with Novozym SP435 for 48 h at 65 ° C, stirring speed 1200 rpm). The reaction was checked by means of thin layer chromatography and the conversion (based on the naringin used) was determined.

sales

4.1 Stearic acid +

4.2 Palmitic acid ++ sales

4.3 Lauric acid ++

4.4 oleic acid +

4.5 Coumaric acid +

4.6 Capric acid +

4.7 Cinnamic acid +

4.8 4- ++

hydroxyphenylacetic

4.9 5-phenylvaleric acid ++

4.10 4-phenylbutyric acid ++

4.11 12-hydroxy stearic acid

4.12 Edenor UKD6010

+ means up to 15% sales ++ means over 15% sales

Example 5: Inhibition of tyrosinase activity

Tyrosinases catalyze an important physiological step in melanin synthesis (L-dopa to L-dopaquinone, which continues to cyclize and is converted to dopachrome again by a tyrosinase). Inhibiting tyrosinase can lead to a skin-lightening effect.

The activity of fungal tyrosinase (Sigma) was different in the presence of

Concentrations of the active ingredients according to the invention based on the enzymatic

Conversion of LDOPA to dopachrome determined. The absorption maximum of the dopachrome (red-brown) is λ = 475nm. The linear increase in absorption (A) of the Dopachromes per unit time (t) is a measure of the activity of tyrosinase (ΔA / Δt). The activity of the tyrosinase in the absence of the active substances (ΔAi / Δti) served as a reference and was set to 100%. The residual tyrosinase activity was determined under analogous conditions in the presence of the active compounds (ΔA ₂ / Δt ₂ ). Each measurement was carried out twice in parallel batches. The fluctuation in the results of the method is approximately ± 10%.

Chemicals used:

L-3,4-dihydroxyphenylalanine (L-DOPA) (Sigma) o KH ₂ PO ₄ (JT Baker)

Tyrosinase, 50,000 units (Sigma)

KOH

Required solutions: 5 50 mM KH ₂ PO ₄ buffer in H ₂ O bidist. (Adjustment to pH = 6.5 with 1 M aqueous KOH)

2.5 mM L-DOPA in H ₂ O bidist.

340 U / ml tyrosinase stock solution in cold KH ₂ PO ₄ buffer, pH = 6.5

Stock solutions of the active ingredient to be tested in bidist. Water or ethanol, in which the concentration of the active ingredient was 10 times higher than indicated in the line "Active ingredient concentration 0 in the test system" under "Results"

Reaction Cocktail:

10 ml KH2PO4 buffer 10 ml L-DOPA 9 ml H ₂ O bidist.

The reaction cocktail, like the tyrosinase stock solution, was only prepared before the start of the experiment. The tyrosinase stock solution must be stored in a cool place. The L-DOPA solutions should be kept dark and in well-sealed containers with the exclusion of oxygen. If the color changes to gray (oxidation by atmospheric oxygen), the solution must be freshly prepared. Test system (sample volume 1 ml) and reaction procedure: 33 μl tyrosinase stock solution 100 μl active ingredient stock solution 5 ad 1000 μl reaction cocktail

The activity of the tyrosinase in the absence of the active ingredients served as a reference and was set to 100%. All samples were mixed thoroughly on the Vibrofix before the start of the measurement. The pH was checked and, if necessary, adjusted to pH = 6.5. The measurement was carried out using a Uvikon 860 photometer from Kontron. The absorption Ό of the dopachrome was at the absorption maximum at λ = 475nm over a period of 5 min. detected at 25 ° C, the measurement interval was 20-30 s.

Results:

Active ingredient: 6-O-naringin (3-phenyl-propionic acid) ester from Example 2 15 active ingredient concentration in the test system 0.005% 0.05% 0.5% (w / v in H ₂ O bidist.) Residual activity of the tyrosinase in% 98.9 69.1 0.7

(IC50 = 0.18%)

Active ingredient: 6-O-naringin (p-CI-phenylacetic acid) ester from Example 3 .0 active ingredient concentration in the test system 0.01% 0.1% (w / v in 98% ethanol)

Residual activity of tyrosinase in% 45.1 15.4

Example 6: Phototoxicity 5

Dermal fibroblasts of human skin were found with increasing concentrations of retinol (Table 1), 6-O-naringin (p-CI-phenylacetic acid) ester (Table 2) and 6-O-naringin (3-phenylpropionic acid) ester (Table 3) cultured. The phototoxicity of the substances was measured using an MTT test. To determine the 0 phototoxicity, the treated cells were irradiated with simulated sunlight, corresponding to a dose of 10 J UV-A / cm2. The vitality of untreated cells was set to 100% and all other values related to it. The cells were irradiated with a sunlight simulator, from whose emission spectrum the UV-A portion of the radiation was measured for quantification. The advantage of this experimental design is the fact that the entire spectrum of sunlight is used and the everyday situation is thus perfectly reproduced. In contrast, many other research laboratories work with pure UV-A and / or UV-B lamps.

Table 1: Phototoxicity of retinol

Concentration retinol (ppm) vitality (%; in brackets: SEM)

0.0028 99 (7.4)

0.014 95 (19.8)

0.028 80 (25.9)

0.14 28 (11, 8)

0.28 4 (2.1)

Table 2: Phototoxicity of 6-O-naringin (p-CI-phenylacetic acid) ester

Concentration of the naringin derivative (ppm) Vitality (%; in brackets: SEM)

5 115 (15)

10 96 (17.2)

50 81 (8.3)

100 3 (1, 7)

500 3 (1, 8)

Table 3: Phototoxicity of 6-O-naringin (3-phenyl-propionic acid) ester concentration of the naringin derivative (ppm) vitality (%; in brackets: SEM)

5 125 (5.8) 10 103 (19.8) 50 98 (15.1) 100 101 (8.3) 500 29 (4.5) 1000 2 (0.5) The results show that 6-0-naringin (3-phenyl-propionic acid) ester and 6-O-naringin (p-CI-phenylacetic acid) ester only have toxic effects in higher concentrations compared to retinol. Retinol is toxic even in very low concentrations. The decrease in vitality by several orders of magnitude confirms the strong phototoxicity of retinol.

Example 7: Effects on light-induced expression of MMP-1, TIMP and Colc mRNA

The effects of 6-O-naringin (3-phenyl-propionic acid) ester (Table 4) and 6-O-naringin (p-CI-phenylacetic acid) ester (Table 5) on the light-induced expression of MMP-1, TIMP and COICH were measured at subphototoxic concentrations. For this, the amount of mRNA for MMP1, TIMP and Colc was quantified. Skin models were treated with the test substances for 12 hours and then irradiated with simulated sunlight, corresponding to a dose of 10 J UV-A / cm ² . After a further 48 hours in the presence of the active ingredients, the RNA of the cells was prepared and analyzed by Northern blots with specific gene probes. To control the amount of RNA used in the experiments, Northern blots were carried out with an 18S-specific gene probe. To quantify the signal intensities, the autoradiograms were measured densitometrically and the values of the signals for MMP1, TIMP and Colch were related to the associated values of the 18S signals. The numerical values in the tables represent the densitometric quantification of the signals of a Northern blot after their normalization. The light-induced expression of MMP1, TIMP and Col _α ι in the case of untreated cells was set to 100% in each case and all other values based on this.

Table 4: Effects of 6-O-naringin (p-CI-phenylacetic acid) ester on the expression of MMP1, TIMP and collagen

Concentration of

Naringin derivative (ppm) MMP1 TIMP collagen

0, unirradiated 100 100 100

0, irradiated 135 89 66

5, irradiated 129 62 56

50, irradiated 40 77 79 Table 5: Effects of 6-0-naringin (3-phenyl-propionic acid) ester on the expression of MMP1, TIMP and collagen

Concentration of

Naringin derivative (ppm) MMP1 TIMP Coll

0, unirradiated 100 100 100 0, irradiated 135 89 66

10, irradiated 167 104 74

100, irradiated 87 134 99

Irradiation of skin models with simulated sunlight resulted in a strong induction of MMP1 mRNA synthesis, while the synthesis of collagen was down-regulated. The production of TIMP remained largely unaffected. Table 4 shows that 50 ppm 6-O-naringin (p-CI-phenylacetic acid) ester very effectively reduced the sunlight-induced expression of MMP-1. The expression of TIMP was only slightly influenced, the expression of Colαi is significantly increased compared to the irradiated, untreated sample. Treatment of the cells with 100 ppm 6-O-naringin (3-phenyl-propionic acid) ester reduced the sunlight-induced expression of MMP-1 to the level of the unirradiated, untreated sample (Table 5). In contrast, the expression of TIMP increased by approx. 35%. The expression of Colαi could be increased to the level of the unirradiated, untreated culture.

The percentage change in the expression of MMP, TIMP and Colαi in cultures of irradiated fibroblasts after treatment with 50 or 100 ppm of the tested naringin derivatives compared to irradiated, untreated cultures is summarized in Table 6.

Table 6:

Expression of 6-O-naringin-6-0-naringin (3-phenyl

(p-CI-phenylacetic acid) ester propionic acid) ester (50 ppm) (100 ppm)

MMP -70% -37% TIMP -15% 50% Colαi 27% 64% The concentration given in Table 6 led to a significant inhibition of MMP expression and an increased Colαi for both naringin derivatives. Production. The 6-O-naringin (3-phenyl-propionic acid) ester increased the TIMP production considerably, while 6-0-naringin (p-CI-phenylacetic acid) ester had little influence.

Example 8: Effect on collagen production

To demonstrate the increased production of collagen at the protein level, fibroblasts were treated with the test substances in a three-dimensional culture system for 5 days. On the sixth day, the amount of collagen formed compared to non-collagen protein was determined by incorporating tritiated protein. Table 7 shows the percentage increase in the proportion of collagen in the total protein, determined from treated fibroblast cultures compared to untreated cultures.

Table 7:

6-O-naringin (3-phenyl- 6-O-naringin

(propionic acid) ester (p-CI-phenylacetic acid) ester

Concentration 1 ppm 10ppm 100ppm 5ppm 50ppm

Increase in

Collagen production 8% 8% 19% -3% 41%

Claims

claims

1. Flavone and isoflavone glycoside derivatives of the general formula (I): [Aι-C (= 0) 0] _m - [XOZ] - [OC (= O) -A ₂ ] _n (I), in which [XOZ] represents a flavone or isoflavone glycoside structure, in which X represents a basic body of flavone or isoflavone of the formula (Ila) or (Ilb)

(Ila) (Ilb)

represents, wherein the (iso-) flavone basic body is mono- or polysubstituted and / or mono- or polysubstituted (hydrogenated), where Z (sugar) stands for a mono-, disaccharide or polysaccharide, which is acetally bound to the radical X and n- is substituted in a ester-like manner with A ₂ , in which [Aι-C (= O)] represents an acyl radical on the base of flavone or isoflavones, in which Ai and A ₂ independently of one another are a polyunsaturated C ₁₅ - C ₂₅ -

Alkenyl radical with at least 4 isolated and / or at least two conjugated

Represent double bonds or an arylaliphatic radical with 1-4 methylene groups between the ester group and aromatic ring, in which [C (= O) A ₂ ] represents an acyl radical on the sugar Z, in which n is an integer (1, 2, 3, .. .), but not 0, where m is an integer including 0 (0, 1, 2, 3, ...), and where R1, R2, R3 represent hydroxyl groups or hydrogen atoms.

2. The derivatives of claim 1, wherein Z is a monosaccharide, in particular rhamnose, threose, erythrose, arabinose, lyxose, ribose, xylose, allose, old rose, galactose, glucose, gulose, idose, mannose, talose and fructose, or a disaccharide, in particular a disaccharide which is built up from the aforementioned monosaccharides, in their naturally occurring stereoisomeric forms.

3. The derivatives of claim 1 or 2, wherein the (iso-) flavone glycoside base X-O-Z in the general formula (I) is asparagine, orientin (lutexin), cisorientin (lutonaretin), isoquercitin, naringin, or rutin.

4. The derivatives of one of the preceding claims, wherein the (iso-) flavone glycoside base X-O-Z naringin of the formula (III)

(III) is.

5. The derivatives of one of the preceding claims, wherein X-O-Z is naringin according to formula (III); where A is the acyl radical of one of the following acids: p-chlorophenylacetic, hydrocinnamon, stearin, 12-hydroxystearin, palmitin, lauric, oil, coumaric, caprin, cinnamon, 4-phenyl butter, 4 -Hydroxyphenylacetic acid, 5-phenylvaleric acid or one of the mixtures available under the trade names Edenor UKD 6010 and UKD 7505; and where n is 1 or 2 and m is 0 at the same time.

6. The derivatives of claim 5, wherein n is 1, m is 0 and A is bound to the primary OH group of the sugar in formula (III).

7. The derivatives of one of claims 1 to 4, wherein X-O-Z is naringin according to formula (III); where A is the acyl residue of one of the following acids: p-chlorophenylacetic, hydrocinnamon, stearin, 12-hydroxystearin, palmitin, lauric, oil, coumaric, capric, cinnamon, 4-phenyl butter, 4 -Hydroxyphenylacetic, 5-phenylvaleric acid or one of the mixtures available under the trade names Edenor UKD 6010 and UKD 7505; and where n is 1 or 2 and m is 1 at the same time.

8. The derivatives of claim 7, wherein n and m are 1, A2 to the primary OH group of the sugar in formula (III) and A _{1 to} either the 5-OH group of the benzopyran or to the 4'-hydroxy Group of the phenyl ring are bound.

9. The derivatives of claim 8, wherein n is 2 and m 1, an A2 to the primary OH group and the second A ₂ to one of the secondary OH groups, in particular to one of the two secondary OH groups on the same or at one of the three secondary OH groups of the second six-membered ring, the sugar in formula (III) and A1 are bound either to the 5-OH group of the benzopyran or to the 4-hydroxy group of the phenyl ring.

10.A process for the preparation of the derivatives of formula (I) from one of the preceding claims, characterized in that an acetal X-O-Z from

Sugar and (iso-) flavone base, these (iso-) flavone base being in the form of the pure substance or as a mixture of plant extracts of different origins, with a polyunsaturated fatty acid with at least four isolated double bonds and / or with at least two conjugated double bonds, with an arylaliphatic carboxylic acid, with an ester thereof

Carboxylic acids or with an activated fatty acid derivative under the catalytic action of one or more enzymes is esterified or transesterified.

11. The method of claim 10, wherein the polyunsaturated fatty acid is a conjugated linoleic acid (octadecadienoic acid).

12. The method of claim 10 or 11, wherein the enzyme or enzymes is one or more hydrolases.

13.The method of claim 12, wherein the hydrolase (s) are the lipases from Candida rugosa (formerly Candida cylindracea), Candida antarctica, Geotrichum candidum, Aspergillus niger, Penicillium roqueforti, Rhizopus arrhizus and Mucor miehei, in particular the lipase (isoenzyme B) from Candida antarctica, is (are).

14. The method of any one of claims 10 to 13, wherein the esterification reaction is followed by a step for purifying the compounds of formula (I), which is either an aqueous two-phase extraction process with organic solvents such as n-hexane, cyclohexane, THF or dieethyl ether or a chromatographic method on silica gel, preferably with ethyl acetate / methanol or dichloromethane / methanol mixtures with small amounts of acetic acid and / or water.

15. Cosmetic or pharmaceutical composition or food or feed composition containing at least one of the derivatives of one of claims 1 to 9.

16. Use of a derivative according to any one of claims 1 to 9 for the cosmetic treatment of sunlight-induced aging of human skin.

17. Use of a derivative according to one of claims 1 to 9 for cosmetic skin lightening.