DE102007020554A1 - Nucleic acid-containing cosmetic and / or pharmaceutical preparations for the treatment of epithelial covering tissue - Google Patents

Abstract

The present invention relates to nucleic acid-containing cosmetic and / or pharmaceutical preparations for the treatment of epithelial covering tissue and to the use of nucleic acids as chemosensitizer for the treatment of hyperplasia epithelial covering tissue.

Description

The The present invention relates to nucleic acid containing cosmetic and / or pharmaceutical preparations for the treatment of epithelial Cover tissue and the use of nucleic acids as Chemosensitizer for the treatment of hyperplasia epithelial covering tissue.

Kanzerosen and precancerous lesions are among the most hyperplastic Diseases of the epidermis and are common indications in dermatology with increasing incidence.

Responsible These are different from individual genetic factors Environmental factors. In addition to viral and chemical influences is the chronic sunlight exposure the main risk. precancerous lesions are typically pre-invasive predecessor lesions of squamous cell carcinoma. The most common precancerous condition is actinic keratosis (also called senile keratosis), which is usually multiple in UV-loaded areas (bald head, forehead, bridge of nose, lips, Back of the hand) occurs. At these predilection sites It can also be a frequent occurrence of basaliomas come. In case of multiple occurrence of precancerous lesions and basaliomas is a surgical excision of the skin lesions both unfavorable for cosmetic as well as pragmatic reasons.

Previous Methods of choice are cryotherapy, local chemotherapy with 5-fluorouracil and topical treatment with imiquimod, one so-called immune response modifier. Especially with the two The latter is a therapy lasting several weeks necessary, in which it is too painful and inflammatory Processes is coming. This is a common reason that the therapy is stopped prematurely and as a result recurrences occur.

A Alternative to the mentioned therapeutic options is therefore desirable.

Surprisingly It has been found that the use of nucleic acids or nucleic acid-containing mixtures apoptosis-promoting on keratinocytes.

object The present invention is therefore a cosmetic and / or pharmaceutical Preparation for the treatment of epithelial covering tissue by it characterized in that it contains nucleic acids.

The contained in the preparation according to the invention Nucleic acids can be natural or be of synthetic origin. You can also hydrolyze, partially hydrolyzed or denatured. Preferably, the Nucleic acids selected from synthetic nucleic acids, Nucleic acids of eukaryotic origin and nucleic acids of bacterial origin, especially among nucleic acids from Escherichia coli and Clostridium perfringens.

preferred contained in the preparation according to the invention Nucleic acids are selected from high molecular weight bacterial DNA, low molecular weight bacterial DNA, high molecular weight eukaryotic DNA, low molecular weight eukaryotic DNA as well under oligonucleotides.

Usable according to the invention Oligonucleotides have a length of 5 to 100, in particular 5 to 70, preferably 10 to 50, preferably 10 to 40 and more particularly preferably from 12 to 30 nucleotides.

Preferably the nucleobases are those in the invention Preparation contained nucleic acids not methylated.

• a) Veränderung der Internukleosidbrücken: Austausch von Phosphodiestern gegen Methylphosphonate, Phosphoramidate, Phosphorothioate (PTO) oder Hydroxylamine;
• b) Veränderung der Zuckerkomponenten: Austausch der Ribose gegen diverse Hexo- bzw. Pentopyranosen oder 3'-5'-carbocyclisch verbrückte Derivate der 2'-Deoxyribose (Steffens R & Leumann CJ: Tricyclo-DNA: A phosphodiesterbackbone based DNA analog exhibiting strong complementary base-pairing properties. J Am Chem Soc; 119: 11548–11549, 1997);
• c) Austausch des Strangrückgrats: Austausch der Polyesterketten auf Basis von Zucker-Phosphat-Einheiten gegen Carboxamidketten auf Basis von Aminosäurederivaten wie N-(2-Aminoethyl)-glycin-Einheiten;

The nucleic acids which can be used according to the invention can be chemically modified completely (all nucleotides) or partially (only a few nucleotides) in a manner known to the person skilled in the art. Preferred modifications are, for example:

• a) Alteration of the internucleoside bridges: Exchange of phosphodiesters for methylphosphonates, phosphoramidates, phosphorothioates (PTO) or hydroxylamines;
• b) Modification of the sugar components: Exchange of the ribose for various hexo- or pentopyranoses or 3'-5'-carbocyclic bridged derivatives of 2'-deoxyribose (Steffens R & Leumann CJ: Tricyclo-DNA: A phosphodiester-based DNA analog exhibiting strong complementary base-pairing properties, J Chem Chem, 119: 11548-11549, 1997);
• c) replacement of the strand backbone: replacement of the polyester chains based on sugar-phosphate units by carboxamide chains based on amino acid derivatives such as N- (2-aminoethyl) -glycine units;

Particularly according to the invention preferred are phosphorothioate phosphodiester mixmers.

in the The scope of the present invention is used under epithelial cover tissue on the one hand, the outer body surface covering skin (consisting of subcutis, corium and epidermis), on the other hand, the hollow organs and body cavities lining tissues, including the epithelia of the Uterus and mouth.

The preparation according to the invention is in particular for the treatment of hyperplastic diseases of the epithelial Cover tissue and in combination with cosmetic agents (eg. As AHA (alpha hydroxy acids)) or UV light for improvement the skin image can be used. In particular, the inventive Oligonucleotides due to their apoptosis-sensitizing effect in combination with cosmetic agents or UV light as anti-aging agents, z. B. for the treatment of age spots, calluses or the so-called Landmann's skin (Cutis rhomboidalis), can be used. By the Apoptosis sensitization of hair keratinocytes can they are also used for the epilation or for the induction of hair loss become.

Hyperplastic Diseases of the epithelial covering tissue are all those diseases, where an excessive proliferation or decreased apoptosis of epithelial cells is present. exemplary for this are cancerous and precancerous as well Verrucae (warts) called.

The inventive preparation is also for the prophylaxis and treatment of various other diseases or undesirable conditions, in particular for the prophylaxis and treatment of psoriasis (especially nail psoriasis, which is usually treated with 5-fluorouracil) as well of cutaneous metastases of epithelial tumors, including the mucosa.

Farther the preparation according to the invention is suitable z. B. in combination with the so-called. "Chemical peeling", to smooth out unevenness of the skin, for example calluses or other thickening of the skin. In this scrub are mostly AHA (alpha hydroxy acids), PHAs (polyhydroxy acids), TCA (Trichloroacetic acid) and glycolic acid.

Preferably, the nucleic acids contained in the preparation according to the invention are present as oligodeoxynucleotides (ODN). These ODNs preferably have one, two or more CpG motifs, as described, for example, in US Pat DE-A-102 33 994.5 of the applicant, to which reference is hereby made in its entirety.

CpG motifs are distinguished by three major classes of CpG ODN: CpG A- (also CpG-D), CpG-B (also CpG-K) and CpG-C-ODN.

The Structure of CpG-A is through a chimeric backbone marked: The 5 'and 3' ends are for increased stability phosphorothioate modified against nucleases, while the middle region consists of unmodified phosphodiester. unmodified Phosphodiester oligonucleotides are in vivo by the body's own Nucleases degraded very rapidly. By chemical modification of the Phosphodiester backbone, the so-called phosphorothioate modification, to achieve increased stability of the ODN (oligo-deoxynucleotide) Nucleases. In the phosphorothioate modification is not an Binding oxygen atom of the phosphate group replaced a sulfur atom. CpG A-ODN usually have only one CpG motif, which is embedded in a palindromic sequence. additionally show at the 5'- and at the 3'-end consequences of guanines, the well for the uptake of the oligonucleotides and the intracellular Localization of importance.

CpG-B ODN have a phosphorothioate backbone and are common several CpG motives. At the 5 'end is often a TCGTCG motif.

CpG ODN C represent a mixture of CpG-A and CpG-B-ODN like the CpG-B ODN have a phosphorothioate backbone frequently several CpG motifs and often at the 5'-end TCGTCG motif. Like the CpG-A ODN, they contain a central CpG motif, which is embedded in a palindromic sequence. They are missing but the guanine consequences.

Further inventively preferred nucleic acids are superstructure-forming ODN, as described for example in the DE-A-103 61 502.4 of the applicant, to which reference is hereby made in its entirety.

Under Superstructure-forming nucleic acid sequences are nucleic acids to understand that are capable of non-covalent bonding to other nucleic acid superstructures, especially G-quadruplexes, to train so-called "Frayed Wires" or "i-motives". Preferably, usable according to the invention Superstructure-forming nucleic acid sequences C, G or I-rich sequences containing C, G or I in the range of 25% to 100%, preferably 50% to 100%, more preferably 75% to 100% and most preferably 100%. In a very special way preferred are poly-I-homopolymers, poly-C-homopolymers or poly-G-homopolymers. Where C is cytosine, G is guanine and I for inosine.

Especially effective are C-, G-, T- and I-rich ODNs (I-rich means inosine rich. Inosine is the hypoxanthine-containing nucleoside). These ODN are particular suitable for forming DNA superstructures. The formation of such ODN superstructures is primarily dependent on the Base composition of ODN. Of further importance in the formation of ODN superstructures are the pH, the ionic strength, the temperature and the presence of certain cations.

Guanosine-rich and also inosine-rich ODNs can via Hoogsteen base pairings Forming G-Quartets that are "stacked" to form of G-quadruplexes. These constitute a multimer of four DNA single strands. The formation can be inter- and intramolecular The quadruplexes are thus one, two or four molecules include. Important here is that the ODN has several consecutive Guanine included. G-quadruplexes can be synthesized by both phosphodiester as well as phosphorothioates.

Next G-quadruplexes, some G-rich oligos can also be called "Frayed Wires. "For this purpose, inter alia, longer G-episodes in ODN necessary. These structures are about them G sequences for the aggregation of ODN, at the very many single strands can be involved.

C-rich Oligonucleotides can be neutral and especially light acidic pH range relatively stable so-called "i-motifs" form. Prerequisite are several episodes of at least two consecutive cytosines. About the Protonation of a cytosine can then be three hydrogen bonds be formed into another non-protonated cytosine. I motifs can be used by both phosphodiester and phosphorothioates be formed.

advantageously, be by the administration of DNA oligonucleotides or DNA bacterial Originating epidermal cells for apoptosis (controlled Sensitized to cell death), so that the concentration of 5-fluorouracil or any other pro-apoptotic stimulus can, which significantly reduces the unwanted side effects. An additional advantage of the invention Use of nucleic acids is that such DNA molecules have an anti-inflammatory effect on skin cells which can relieve the side effects of local chemotherapy.

apoptosis describes a physiological process in which it is controlled Downfall of cells is coming. Disorders in regulation Of apoptosis often occur in the context of hyperproliferative diseases such as B. Cancer. The protein kinase Akt (also PKB or RAC) plays this often plays a central role as it pro-apoptotic Signal paths, as a result z. For activation of caspases comes, hinders. In different cancers, a constitutive Activation of act can be observed. A suppression of Akt activity Here, it is desirable to allow cells to enter apoptosis to enable.

Surprisingly the applicant could show that very different DNA species cell-specific for epidermal keratinocytes the Akt activity inhibit. Both CpG motif-containing (CpG-1 PTO) and DNA oligonucleotides without CpG motif (non-CpG-5-PTO) show here a concentration-dependent inhibition the basal phosphorylation of the two functional phosphorylation sites of Akt (serine 473, threonine 308) after an incubation of 30 minutes (see Example 1). Furthermore it could be shown that 4 μM CpG-1 PTO leads to a permanent inhibition of Akt of at least 24h (see example 2). This is therapeutically interesting, as a result a relatively long time window of Akt inhibition is given.

By Deletion mutants of CpG-1-PTO and non-CpG-5-PTO could be the minimum length the DNA ODN causing suppression of Akt (see examples 3 and 4). For deletion mutants of CpG-1-PTO could have a length of ≥ 12 bases as minimum length be determined at a concentration of 4 μM. For Deletion mutants of non-CpG-5-PTO showed a jumpy Suppression of act from a length of> 12 bases at a concentration of 4 μM.

In addition to synthetic DNA ODN, natural DNAs and DNA-containing products have also been considered Akt suppression in skin keratinocytes (see Example 5). It could be shown that high-molecular DNA from Escherichia coli (ICN) and Clostridium perfringens (ICN) suppresses the basal Akt activity. Already 3.2 μg / ml DNA from C. perfringens are sufficient for a complete suppression, DNA from E. coli showed a partial suppression of the Akt phosphorylation at 3.2 μg / ml.

low molecular weight Salmon sperm DNA (ICN) showed no effect on Akt Ineffective in this context was an autolysate of enterococci (Symbio Flor 1, SymbioPharm GmbH, Herborn) and the mycobacterial Extract BCG (Bacillus Calmette Guerin, BCG-medac, Medac GmbH, Hamburg).

Further was investigated to what extent the suppression of Akt by DNA general principle or if there are cell-specific effects. For this purpose, different cell species were used, each with 4 μM CpG-1 PTO or non-CpG-5 PTO incubated for 30 minutes. Surprisingly showed that only epidermal keratinocytes (NHK = normal human keratinocytes - not shown, HaCaT, A-431) a suppression of act to respond to the administration of oligonucleotides (see example 6). The applied DNA had no effect on Akt in SZ95 (sebocyte line), Cos-7 (renal epithelial cell line), primary Skin fibroblasts and G361 (melanoma cell line). It is therefore to be assumed that the suppression of act and the related effects on the apoptosis behavior is a keratinocyte-specific phenomenon are.

Further was checked if keratinocytes mediated by the DNA Suppression of Akt can be sensitized to apoptosis. HaCaT keratinocytes were treated with CpG-1-PTO or non-CpG-5-PTO and an established apoptosis inducer (staurosporine) for Incubated for 24 hours. Subsequently, the caspases were in the Western blot 3 and 8, and their degradation products detected. Form caspases a family of proteases that in the context of apoptosis to a directed proteolysis of cell proteins. once activated so-called initiator caspases (eg caspase 8) cut the downstream effector caspases (eg caspase 3) and activate it. The activated effector caspases can then cellular structural proteins, cell cycle proteins and Digest kinases. In this example, the addition of Staurosporine to a concentration-dependent fragmentation of Caspase 8 and 3. When the cells are co-administered with CpG-1-PTO or non-CpG-5-PTO is the apoptotic effect reinforced by staurosporine. In that sense, the used DNA ODN as "apoptosis sensitizer" (see Example 7).

The Effectiveness of the used oligos was further investigated with respect to cytochrome C release (see example 8th). Cytochrome C is the largest in vital cells Part in the mitochondria. Only in the context of apoptosis it comes to an accumulation of cytoplasmic cytochrome C. There acts Cytochrome C acts as an activator of caspase 9 and is thus a very early stage Apoptosis markers. The data presented prove that it is caused by CpG-1 PTO and non-CpG-5 PTO for increased release of Cytochrome C enters the cytoplasm and thus enters apoptosis is relieved.

The The effectiveness of the oligos in terms of apoptosis is cell-specific, neither human sebocytes (see Example 9) nor fibroblasts (see example 10) show an increased degradation of caspases by the Use of oligos.

As shown in Examples 3 and 4, the suppression of Akt depends of the chain length of the used oligos. To show, that this parameter is also crucial for the Effect as "apoptosis sensitizer" has been the activity of Caspase 8 after application of short deletion mutants (hexamers) of CpG-1 PTO and non-CpG-5 PTO. It could be shown, that only the long polymers with a chain length of Twenty nucleotides (CpG-1-PTO, non-CpG-5-PTO) degraded caspase 8, while the hexamers (CpG-9-PTO, n-CpG-5G-PTO) do not lead to increased apoptosis (see Example 11).

Around also here to complete the picture, was also the content determined on cytoplasmic cytochrome C. Affirmative it was found that n-CpG-5-PTO enhances apoptosis, while the short n-CpG-5G PTO has no significant effect on cytochrome C (see Example 12).

DNA oligo-nucleotides Currently, they are predominantly used as antisense molecules tested. Because some of these molecules also have one or more CpG motives include, should be tested, whether a commercial Oligo with these characteristics (G3139, Oblimersen, Genasense) a basal Induces apoptosis. G3139 is a DNA 18mer with 2 CpG motifs, originally designed as a bcl-2 antisense molecule and so should increase apoptosis in malignant cells. We were able to show that treatment of human keratinocytes (HaCaT, A-431) with G3139 PTO - without additional proapoptotic stimulus - to no induction of Caspases leads (see Example 13).

Around the effect of oligonucleotides as an apoptosis sensitizer on to characterize three different classes of CpG ODN (CpG A ODN also known as CpG D ODN, CpG B ODN also as CpG K ODN known and CpG-C-ODN). The structure of CpG-A is through a chimeric backbone featured: the 5 'and 3 'ends are for increased stability against nucleases phosphorothioate-modified, while the middle region consists of unmodified phosphodiesters. Unmodified phosphodiester oligonucleotides become very rapid in vivo by endogenous nucleases reduced. By chemical modification of the phosphodiester backbone, the so-called phosphorothioate modification, one achieves an increased stability of the ODN (oligo-deoxynucleotide) towards nucleases. In the phosphorothioate modification is not involved in the binding Replaced oxygen atom of the phosphate group by a sulfur atom. CpG-A ODN usually have only one CpG motif in a palindromic sequence is embedded. In addition they point at the 5'- and the 3'-end Followed by guanines, probably for the inclusion of Oligonucleotides and the intracellular localization of Meaning are. As a typical representative of CpG-A-ODN was the Sequence 1585 (A) tested.

CpG-B ODN have a phosphorothioate backbone and are common several CpG motives. At the 5 'end is often a TCGTCG motif. As a typical representative of CpG-B-ODN was the above-mentioned Tested sequence of CpG-1 PTO.

CpG ODN C represent a mixture of CpG-A and CpG-B-ODN like the CpG-B ODN have a phosphorothioate backbone frequently several CpG motifs and often at the 5'-end TCGTCG motif. Like the CpG-A ODN, they contain a central CpG motif, which is embedded in a palindromic sequence. They are missing but the guanine consequences. As a typical representative of CpG-A-ODN was tested the sequence M-363-C. To further characterize the ODN effect were 20-mers of poly-G (n-CpG-6-PTO), poly-T (n-CpG-3-PTO), and as a control, the above-mentioned poly-C (non-CpG-5-PTO) tested. The mentioned ODN-PTO became concentration-dependent examined for their effect as an apoptosis sensitizer. there became a later apoptotic marker, the Histone-associated proof Considered DNA fragments. It could be found that CpG-1-PTO (= CpG-B-ODN), non-CpG-5-PTO, n-CpG-3-PTO a marked concentration-dependent Super-induction of apoptosis in the presence of staurosporine effect (see Example 14). n-CpG-6-PTO had the opposite effect, namely a concentration-dependent inhibition of Apoptosis in the presence of staurosporine (see Example 15). M-362-C as a representative of CpG-C-ODN showed no effect on the Apoptosis induction (see Example 15) during, 1585 (A) (= CpG-A-ODN) showed a two-fold effect: at low concentrations (1-4 μM), apoptosis was superinduced, high Concentrations 8 and 16 μM were similar like n-CpG-6 PTO, an inhibition of apoptosis (see Example 16).

The Effectiveness of Nucleic Acids in Formulations for Use especially on the skin depends on the availability of the nucleic acids in the living cells of the skin. The Penetration of a macromolecule through the stratum corneum (natural barrier of the skin) in the skin is not always guaranteed. Liposomes packed nucleic acids however, may penetrate the stratum corneum of skin models. Preparations preferred according to the invention are therefore, those which are the inventively employable nucleic acids contained in liposomes. Equally preferred are preparations containing other suitable carrier systems, eg. B. Nanotechnology-based systems or penetration enhancers (eg, urea, Azone or DMSO).

Particular preference is given to the preparation of suitable liposomes as in DE-A-197 40 092 to which reference is hereby incorporated by reference.

One Another object of the present invention is the use of nucleic acids as a chemosensitizer for the treatment of Hyperplasia epithelial covering tissue.

One Another object of the present invention is a method for the preparation of a cosmetic and / or pharmaceutical preparation for the treatment of hyperplasia epithelial covering tissue, thereby characterized in that one has nucleic acids as for the preparations according to the invention are described, with pharmacologically acceptable and compatible carriers mixed.

The Nucleic acids which can be used according to the invention are in the context of the present invention preferably as a component incorporated into a cosmetic and / or pharmaceutical preparation.

In a preferred embodiment, the preparation according to the invention contains the nucleic acids which can be used according to the invention and additionally at least one apoptosis-inducing substance, in particular 5-fluorouracil or an imidazoquinoline, such as. Eg imiquimod.

The Nucleic acids which can be used according to the invention can also be used in combination with other substances or Therapy forms are used, in particular with those that are selected under: coal tar, phototherapy, photodynamic Therapy (especially with 5-aminolevulinic acid = ALA), Anthralin, cryotherapy and retinoids (ATRA = all-trans-retinoic acid, acitretin, tazarotene)

The Nucleic acids which can be used according to the invention can coincide or time-shift with the other substances or forms of therapy are used. Preferred is the same time Application.

The Dosage and duration of use of the present invention Nucleic acids can be determined by one of ordinary skill in the art adapted and varied.

The cosmetic and / or pharmaceutical according to the invention Preparations, such as creams, gels, lotions, alcoholic and aqueous / alcoholic solutions, emulsions, Wax / fat masses, stick preparations, powders or ointments can - ever by type of formulation - as auxiliaries and additives mild surfactants, oil bodies, emulsifiers, superfatting agents, Bodying agents, thickeners, polymers, silicone compounds, Fats, waxes, stabilizers, biogenic agents, film formers, Swelling agents, UV protection factors, antioxidants, hydrotropes, Preservatives, solubilizers and the like included.

typical Examples of suitable milds, d. H. particularly skin-friendly Surfactants are fatty alcohol polyglycol ether sulfates, monoglyceride sulfates, Mono- and / or dialkyl sulfosuccinates, fatty acid isethionates, Fatty acid sarcosinates, fatty acid taurides, fatty acid glutamates, α-olefinsulfonates, Ether carboxylic acids, alkyl oligoglucosides, fatty acid glucamides, Alkylamidobetaines and / or protein fatty acid condensates, the latter preferably based on wheat proteins.

Examples of oil bodies are Guerbet alcohols based on fatty alcohols having 6 to 18, preferably 8 to 10 carbon atoms, esters of linear C ₆ -C ₂₂ fatty acids with linear C ₆ -C ₂₂ fatty alcohols, esters of branched C ₆ -C ₁₃ carboxylic acids with linear C ₆ -C ₂₂ fatty alcohols, such as. B. myristyl myristate, myristyl palmitate, myristyl stearate, Myristylisostearat, myristyl, Myristylbehenat, Myristylerucat, cetyl myristate, cetyl palmitate, cetyl stearate, Cetylisostearat, cetyl oleate, cetyl behenate, Cetylerucat, Stearylmyristat, stearyl palmitate, stearyl stearate, Stearylisostearat, stearyl oleate, stearyl behenate, Stearylerucat, isostearyl, isostearyl palmitate, Isostearylstearat, isostearyl isostearate, Isostearyloleat, isostearyl behenate, Isostearyloleat, oleyl myristate, oleyl palmitate, oleyl stearate, oleyl isostearate, oleyl oleate, Oleylbehenat, oleyl erucate, behenyl myristate, behenyl palmitate, behenyl, Behenylisostearat, behenyl oleate, behenyl behenate, behenyl erucate, erucyl myristate, erucyl, erucyl, erucyl, erucyl oleate, erucyl behenate and erucyl consideration.

In addition, esters of linear C ₆ -C ₂₂ fatty acids with branched alcohols, in particular 2-ethylhexanol, esters of hydroxycarboxylic acids with linear or branched C ₆ -C ₂₂ fatty alcohols, in particular dioctyl malates, esters of linear and / or branched fatty acids with polyhydric alcohols (such as, for example, propylene glycol, dimerdiol or trimer triol) and / or Guerbet alcohols, triglycerides based on C ₆ -C ₁₀ fatty acids, liquid mono- / di- / triglyceride mixtures based on C ₆ -C ₁₈ fatty acids, esters of C ₆ -C ₂₂ fatty alcohols and / or Guerbet alcohols with aromatic carboxylic acids, in particular benzoic acid, esters of C ₂ -C ₁₂ dicarboxylic acids with linear or branched alcohols having 1 to 22 carbon atoms or polyols having 2 to 10 carbon atoms and 2 to 6 hydroxyl groups , vegetable oils, branched primary alcohols, substituted cyclohexanes, linear and branched C ₆ -C ₂₂ fatty alcohol carbonates, Guerbet carbonates, esters of benzoic acid with linear and / or branched C ₆ -C ₂₂ -alcohols (eg. B. Finsolv ^® TN), linear or branched, symmetrical or asymmetrical dialkyl ethers having 6 to 22 carbon atoms per alkyl group, ring-opening products of epoxidized fatty acid esters with polyols, silicone oils and / or aliphatic or naphthenic hydrocarbons, such as. As squalane, squalene or dialkylcyclohexanes.

• (1) Anlagerungsprodukte von 2 bis 30 Mol Ethylenoxid und/oder 0 bis 5 Mol Propylenoxid an lineare Fettalkohole mit 8 bis 22 C-Atomen, an Fettsäuren mit 12 bis 22 C-Atomen, an Alkylphenole mit 8 bis 15 C-Atomen in der Alkylgruppe sowie Alkylamine mit 8 bis 22 Kohlenstoffatomen im Alkylrest;
• (2) C_12/18-Fettsäuremono- und -diester von Anlagerungsprodukten von 1 bis 30 Mol Ethylenoxid an Glycerin;
• (3) Glycerinmono- und -diester und Sorbitanmono- und -diester von gesättigten und ungesättigten Fettsäuren mit 6 bis 22 Kohlenstoffatomen und deren Ethylenoxidanlagerungsprodukte;
• (4) Alkyl- und/oder Alkenylmono- und -oligoglycoside mit 8 bis 22 Kohlenstoffatomen im Alk(en)ylrest und deren ethoxylierte Analoga;
• (5) Anlagerungsprodukte von 15 bis 60 Mol Ethylenoxid an Ricinusöl und/oder gehärtetes Ricinusöl;
• (6) Polyol- und insbesondere Polyglycerinester;
• (7) Anlagerungsprodukte von 2 bis 15 Mol Ethylenoxid an Ricinusöl und/oder gehärtetes Ricinusöl;
• (8) Partialester auf Basis linearer, verzweigter, ungesättigter bzw. gesättigter C_6/22-Fettsäuren, Ricinolsäure sowie 12-Hydroxystearinsäure und Glycerin, Polyglycerin, Pentaerythrit, Dipentaerythrit, Zuckeralkohole (z. B. Sorbit), Alkylglucoside (z. B. Methylglucosid, Butylglucosid, Laurylglucosid) sowie Polyglucoside (z. B. Cellulose);
• (9) Mono-, Di- und Trialkylphosphate sowie Mono-, Di- und/oder Tri-PEG-alkylphosphate und deren Salze;
• (10) Wollwachsalkohole;
• (11) Polysiloxan-Polyalkyl-Polyether-Copolymere bzw. entsprechende Derivate;
• (12) Mischester aus Pentaerythrit, Fettsäuren, Citronensäure und Fettalkohol gemäß DE 1165574 PS und/oder Mischester von Fettsäuren mit 6 bis 22 Kohlenstoffatomen, Methylglucose und Polyolen, vorzugsweise Glycerin oder Polyglycerin,
• (13) Polyalkylenglycole sowie
• (14) Glycerincarbonat.

Examples of suitable emulsifiers are nonionic surfactants from at least one of the following groups:

• (1) addition products of 2 to 30 moles of ethylene oxide and / or 0 to 5 moles of propylene oxide to linear fatty alcohols having 8 to 22 carbon atoms, to fatty acids having 12 to 22 carbon atoms, to alkylphenols having 8 to 15 carbon atoms in the Alkyl group and alkylamines having 8 to 22 carbon atoms in the alkyl radical;
• (2) C _12/18 fatty acid _mono- and diesters of addition products of 1 to 30 moles of ethylene oxide with glycerol;
• (3) glycerol mono- and diesters and sorbitan mono- and diesters of saturated and unsaturated fatty acids having 6 to 22 carbon atoms and their ethylene oxide addition products;
• (4) alkyl and / or alkenyl mono- and oligoglycosides having 8 to 22 carbon atoms in the alk (en) yl radical and their ethoxylated analogs;
• (5) addition products of 15 to 60 moles of ethylene oxide with castor oil and / or hydrogenated castor oil;
• (6) polyol and especially polyglycerol esters;
• (7) addition products of 2 to 15 moles of ethylene oxide with castor oil and / or hydrogenated castor oil;
• (8) _{partial esters} based on linear, branched, unsaturated or saturated C _6/22 fatty acids, ricinoleic acid and 12-hydroxystearic acid and glycerol, polyglycerol, pentaerythritol, dipentaerythritol, sugar alcohols (for example sorbitol), alkyl glucosides (eg. Methylglucoside, butylglucoside, laurylglucoside) as well as polyglucosides (eg cellulose);
• (9) mono-, di- and trialkyl phosphates and mono-, di- and / or tri-PEG-alkyl phosphates and their salts;
• (10) wool wax alcohols;
• (11) polysiloxane-polyalkyl-polyether copolymers or corresponding derivatives;
• (12) mixed esters of pentaerythritol, fatty acids, citric acid and fatty alcohol according to DE 1165574 PS and / or mixed esters of fatty acids having 6 to 22 carbon atoms, methyl glucose and polyols, preferably glycerol or polyglycerol,
• (13) Polyalkylene glycols as well
• (14) Glycerol carbonate.

The Addition products of ethylene oxide and / or of propylene oxide Fatty alcohols, fatty acids, alkylphenols, glycerol mono- and diesters and sorbitan mono- and diesters of fatty acids or to castor oil known, commercially available Products.

These are homolog mixtures whose average degree of alkoxylation ent speaks the ratio of the amounts of ethylene oxide and / or propylene oxide and substrate with which the addition reaction is carried out. C _12/18 fatty acid _mono- and diesters of addition products of ethylene oxide with glycerol are made DE 2024051 PS known as a refatting agent for cosmetic preparations.

alkyl and / or alkenyl mono- and oligoglycosides, their preparation and their use is known in the art. Your production takes place in particular by reaction of glucose or oligosaccharides with primary alcohols with 8 to 18 carbon atoms. In terms of of the glycoside residue is considered to be both monoglycosides in which cyclic sugar residue is glycosidically linked to the fatty alcohol, as well as oligomeric glycosides with a degree of oligomerization bis preferably about 8 are suitable. The degree of oligomerization is while a statistical mean, the one for such technical products usual homolog distribution lies.

Typical examples of suitable polyglycerol esters are Polyglyceryl-2 Dipolyhydroxystearate (Dehymuls ^® PGPH), Polyglycerin-3-Diisostearate (Lameform ^® TGI), Polyglyceryl-4 Isostearate (Isolan ^® GI 34), Polyglyceryl-3 Oleate, Diisostearoyl Polyglyceryl-3 Diisostearate (Isolan ^® PDI), Polyglyceryl-3 methylglucose Distearate (Tego Care ^® 450), Polyglyceryl-3 Beeswax (Cera Bellina ^®), Polyglyceryl-4 Caprate (polyglycerol Caprate T2010 / 90), Polyglyceryl-3 Cetyl ether (Chimexane ^® NL), Polyglyceryl -3 Distearate (Cremophor ^® GS 32) and Polyglyceryl polyricinoleates (Admul ^® WOL 1403), polyglyceryl dimerates Isostearate and mixtures thereof.

Furthermore, zwitterionic surfactants can be used as emulsifiers. Zwitterionic surfactants are those surface-active compounds which carry at least one quaternary ammonium group and at least one carboxylate and one sulfonate group in the molecule. Particularly suitable zwitterionic surfactants are the so-called betaines such as the N-alkyl-N, N-dimethylammoniumglycinate, for example Kokosalkyldimethylammoniumglycinat, N-acylaminopropyl-N, N-dimethylammoniumglycinate, for example Kokosacylaminopropyldimethylammoniumglycinat, and 2-alkyl-3-carboxylmethyl-3-hydroxyethylimidazoline each having 8 to 18 carbon atoms in the alkyl or acyl group and the Kokosacylaminoethylhydroxyethylcarboxymethylglycinat. Particularly preferred is the fatty acid amide derivative known under the CTFA name Cocamidopropyl Betaine. Also suitable emulsifiers are ampholytic surfactants. Ampholytic surfactants are understood as meaning those surface-active compounds which, apart from a C _8/18 -alkyl or -acyl group in the molecule, contain at least one free amino group and at least one -COOH or -SO ₃ H group and are capable of forming internal salts. Examples of suitable ampholytic surfactants are N-alkylglycines, N-alkylpropionic acids, N-alkylaminobutyric acids, N-alkyliminodipropionic acids, N-hydroxyethyl-N-alkylamidopropylglycines, N-alkyltaurines, N-alkylsarcosines, 2-alkylaminopropionic acids and alkylaminoacetic acids each having about 8 to 18 C Atoms in the alkyl group. Particularly preferred ampholytic surfactants are N-cocoalkylaminopropionate, cocoacylaminoethylaminopropionate and C _12/18 acylsarcosine. In addition to the ampholytic also come Quaternary Emulga In particular, those of the esterquat type, preferably methyl-quaternized difatty acid triethanolamine ester salts, are particularly preferred.

As superfatting agent may contain substances such as lanolin and lecithin as well as polyethoxylated or acylated lanolin and lecithin derivatives, Polyol fatty acid esters, monoglycerides and fatty acid alkanolamides be used, the latter also as foam stabilizers serve.

When Bodying agents are primarily fatty alcohols or hydroxy fatty alcohols with 12 to 22 and preferably 16 to 18 carbon atoms and next to it Partial glycerides, fatty acids or hydroxy fatty acids in Consideration. Preference is given to a combination of these substances with alkyl oligoglucosides and / or fatty acid N-methylglucamiden same chain length and / or polyglycerol poly-12-hydroxy stearates.

Suitable thickening agents are, for example, Aerosil types (hydrophilic silicic acids), polysaccharides, especially xanthan gum, guar guar, agar agar, alginates and tyloses, carboxymethyl cellulose and hydroxyethyl cellulose, also higher molecular weight polyethylene glycol mono- and diesters of fatty acids, polyacrylates, (eg. B. Carbopole ^® from Goodrich or Synthalene ^® from Sigma), polyacrylamides, polyvinyl alcohol and polyvinylpyrrolidone, surfactants such as ethoxylated fatty acid glycerides, esters of fatty acids with polyols such as pentaerythritol or trimethylolpropane, fatty alcohol ethoxylates with narrow homolog distribution or Alkyloligoglucoside and electrolytes such as sodium chloride and ammonium chloride.

Suitable cationic polymers are, for example, cationic cellulose derivatives, such as. Example, a quaternized hydroxyethylcellulose obtainable under the name Polymer JR 400 ^® from Amerchol, cationic starch, copolymers of diallyl ammonium salts and acrylamides, quaternized vinylpyrrolidone / vinylimidazole polymers, such. As Luviquat ^® (BASF), condensation products of polyglycols and amines, quaternized collagen polypeptides, for example lauryldimonium hydroxypropyl hydrolyzed collagen (Lamequat® ^® L / Grunau), quaternized wheat polypeptides, polyethyleneimine, cationic silicone polymers such. B. amidomethicones, copolymers of adipic acid and Dimethylaminohydroxypropyldiethylentriamin (Cartaretine ^® / Sandoz), copolymers of acrylic acid with dimethyldiallylammonium chloride (Merquat ^® 550 / Chemviron), polyamino polyamides, such as. B. described in the FR 2252840 A and their crosslinked water-soluble polymers, cationic chitin derivatives such as quaternized chitosan, optionally microcrystalline distributed, condensation products of dihaloalkylene, such as. B. dibromobutane with bis-dialkylamines, such as. B. bis-dimethylamino-1,3-propane, cationic guar gum, such as. ^Jaguar® CBS, ^Jaguar® C-17, ^Jaguar® C-16 from Celanese, quaternized ammonium salt polymers, e.g. B. Mirapol ^® A-15, Mirapol ^® AD-1, Mirapol ^® AZ-1 from Miranol.

When anionic, zwitterionic, amphoteric and nonionic polymers for example, vinyl acetate / crotonic acid copolymers, Vinyl pyrrolidone / vinyl acrylate copolymers, vinyl acetate / butyl maleate / isobornyl acrylate copolymers, Methyl vinyl ether / maleic anhydride copolymers and their Esters, uncrosslinked and polyols crosslinked polyacrylic acids, Acrylamidopropyltrimethylammonium chloride / acrylate copolymers, octylacrylamide / methylmethacrylate / tert. Butylaminoethyl methacrylate / 2-Hydroxyproyl methacrylate copolymers, Polyvinylpyrrolidone, vinylpyrrolidone / vinylacetate copolymers, vinylpyrrolidone / dimethylaminoethylmethacrylate / vinylcaprolactam terpolymers and optionally derivatized cellulose ethers and silicones in question.

Suitable silicone compounds are, for example, dimethylpolysiloxanes, methylphenylpolysiloxanes, cyclic silicones and amino, fatty acid, alcohol, polyether, epoxy, fluorine, glycoside and / or alkyl-modified silicone compounds which may be both liquid and resin-form at room temperature. Also suitable are simethicones, which are mixtures of dimethicones having an average chain length of from 200 to 300 dimethylsiloxane units and hydrogenated silicates. A detailed overview of suitable volatile silicones is also available from Todd et al. in Cosm. Toil. 91, 27 (1976) ,

typical Examples of fats are glycerides, as waxes come u. a. natural waxes, such. Candelilla wax, carnauba wax, Japan wax, esparto wax, cork wax, guaruma wax, rice germ oil wax, Sugarcane wax, ouricury wax, montan wax, beeswax, shellac wax, Spermaceti, lanolin (wool wax), crepe fat, ceresin, ozokerite (Earthwax), petrolatum, paraffin waxes, microwaxes; chemically modified Waxes (hard waxes), such as. Montan ester waxes, Sasol waxes, hydrogenated Jojoba waxes and synthetic waxes, such as. B. polyalkylene waxes and polyethylene glycol waxes in question.

When Stabilizers can be metal salts of fatty acids, such as For example, magnesium, aluminum and / or zinc stearate or ricinoleate be used.

Under biogenic agents are for example tocopherol, tocopherol acetate, Tocopherol palmitate, ascorbic acid, deoxyribonucleic acid, Retinol, bisabolol, allantoin, phytantriol, panthenol, AHA acids, amino acids, Ceramides, pseudoceramides, essential oils, plant extracts and vitamin complexes.

common Film formers are, for example, chitosan, microcrystalline chitosan, quaternized chitosan, polyvinylpyrrolidone, vinylpyrrolidone-vinyl acetate copolymers, Polymers of the acrylic acid series, quaternary cellulose derivatives, Collagen, hyaluronic acid or its salts and the like Links.

Suitable swelling agents for aqueous phases are montmorillonites, clay minerals, pemulen and alkyl-modified carbopol types (Goodrich). Other suitable polymers or swelling agents can the overview of R. Lochhead in Cosm. Toil. 108, 95 (1993) be removed.

• • 3-Benzylidencampher bzw. 3-Benzylidennorcampher und dessen Derivate, z. B. 3-(4-Methylbenzyliden)campher wie in der EP 0693471 B1 beschrieben;
• • 4-Aminobenzoesäurederivate, vorzugsweise 4-Dimethylamino)benzoesäure-2-ethylhexylester, 4-(Dimethylamino)benzoesäure-2-octylester und 4-(Dimethylamino)benzoesäureamylester;
• • Ester der Zimtsäure, vorzugsweise 4-Methoxyzimtsäure-2-ethylhexylester, 4-Methoxyzimtsäurepropylester, 4-Methoxyzimtsäureisoamylester 2-Cyano-3,3-phenylzimtsäure-2-ethylhexylester (Octocrylene);
• • Ester der Salicylsäure, vorzugsweise Salicylsäure-2-ethylhexylester, Salicylsäure-4-isopropylbenzylester, Salicylsäurehomomenthylester;
• • Derivate des Benzophenons, vorzugsweise 2-Hydroxy-4-methoxybenzophenon, 2-Hydroxy-4-methoxy-4'-methylbenzophenon, 2,2'-Dihydroxy-4-methoxybenzophenon;
• • Ester der Benzalmalonsäure, vorzugsweise 4-Methoxybenzmalonsäuredi-2-ethylhexylester;
• • Triazinderivate, wie z. B. 2,4,6-Trianilino-(p-carbo-2'-ethyl-1'-hexyloxy)-1,3,5-triazin und Octyl Triazon, wie in der EP 0818450 A1 beschrieben oder Dioctyl Butamido Triazone (Uvasorb^® HEB);
• • Propan-1,3-dione, wie z. B. 1-(4-tert.Butylphenyl)-3-4'methoxyphenyl)propan-1,3-dion;
• • Ketotricyclo(5.2.1.0)decan-Derivate, wie in der EP 0694521 B1 beschrieben.

Under UV sun protection factors are, for example, at room temperature, liquid or crystalline organic substances present (sunscreen) to understand that are able to absorb ultraviolet rays and the absorbed energy in the form of longer-wave radiation, eg. B. to give off heat again. UVB filters can be oil-soluble or water-soluble. As oil-soluble substances z. To name, for example:

• • 3-Benzylidencampher or 3-Benzylidennorcampher and its derivatives, eg. B. 3- (4-methylbenzylidene) camphor as in EP 0693471 B1 described;
• 4-aminobenzoic acid derivatives, preferably 4-dimethylamino) benzoic acid 2-ethylhexyl ester, 4- (dimethylamino) benzoic acid 2-octyl ester and 4- (dimethylamino) benzoic acid amyl ester;
• Esters of cinnamic acid, preferably 4-methoxycinnamic acid 2-ethylhexyl ester, 4-methoxycinnamic acid propyl ester, 4-methoxycinnamic acid isoamyl ester 2-cyano-3,3-phenylcinnamic acid 2-ethylhexyl ester (octocrylene);
• Esters of salicylic acid, preferably 2-ethylhexyl salicylate, 4-isopropylbenzyl salicylate, homomenthyl salicylate;
• Derivatives of benzophenone, preferably 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone;
• Esters of benzalmalonic acid, preferably di-2-ethylhexyl 4-methoxybenzmalonate;
• Triazine derivatives, such as. 2,4,6-trianilino- (p-carbo-2'-ethyl-1'-hexyloxy) -1,3,5-triazine and octyl triazone, as described in U.S.P. EP 0818450 A1 Dioctyl Butamido Triazone or described (Uvasorb HEB ^®);
• • Propane-1,3-dione, such as B. 1- (4-tert-butylphenyl) -3-4'-methoxyphenyl) propane-1,3-dione;
• • Ketotricyclo (5.2.1.0) decane derivatives as described in U.S. Pat EP 0694521 B1 described.

• • 2-Phenylbenzimidazol-5-sulfonsäure und deren Alkali-, Erdalkali-, Ammonium-, Alkylammonium-, Alkanolammonium- und Glucammoniumsalze;
• • Sulfonsäurederivate von Benzophenonen, vorzugsweise 2-Hydroxy-4-methoxybenzophenon-5-sulfonsäure und ihre Salze;
• • Sulfonsäurederivate des 3-Benzylidencamphers, wie z. B. 4-(2-Oxo-3-bornylidenmethyl)benzolsulfonsäure und 2-Methyl-5-(2-oxo-3-bornyliden)-sulfonsäure und deren Salze.

Suitable water-soluble substances are:

• 2-phenylbenzimidazole-5-sulfonic acid and its alkali, alkaline earth, ammonium, alkylammonium, alkanolammonium and glucammonium salts;
• Sulfonic acid derivatives of benzophenones, preferably 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and its salts;
• Sulfonic acid derivatives of 3-Benzylidencamphers, such as. B. 4- (2-oxo-3-bionylidenemethyl) benzenesulfonic acid and 2-methyl-5- (2-oxo-3-bomylidene) sulfonic acid and salts thereof.

As a typical UV-A filter in particular derivatives of benzoylmethane are suitable, such as 1- (4'-tert-butylphenyl) -3- (4'-methoxyphenyl) propane-1,3-dione, 4-tert-butyl 4'-methoxydibenzoylmethane (Parsol 1789), 1-phenyl-3- (4'-isopropylphenyl) -propane-1,3-dione, and enamine compounds as described in U.S.P. DE 19712033 A1 (BASF). Of course, the UV-A and UV-B filters can also be used in mixtures. In addition to the soluble substances mentioned, insoluble photoprotective pigments, namely finely dispersed metal oxides or salts, are also suitable for this purpose. Examples of suitable metal oxides are in particular zinc oxide and titanium dioxide and, in addition, oxides of iron, zirconium, silicon, manganese, aluminum and cerium and mixtures thereof. As salts silicates (talc), barium sulfate or zinc stearate can be used. The oxides and salts are used in the form of the pigments for skin-care and skin-protecting emulsions and decorative cosmetics. The particles should have an average diameter of less than 100 nm, preferably between 5 and 50 nm and in particular between 15 and 30 nm. They may have a spherical shape, but it is also possible to use those particles which have an ellipsoidal or otherwise deviating shape from the spherical shape. The pigments can also be surface-treated, ie hydrophilized or hydrophobized. Typical examples are coated titanium dioxides, such as. Example, titanium dioxide T 805 (Degussa) or Eusolex ^® T2000 (Merck). Suitable hydrophobic coating agents are in particular silicones and in particular trialkoxyoctylsilanes or simethicones. In sunscreens, so-called micro- or nanopigments are preferably used. Preferably, micronized zinc oxide is used. Further suitable UV sunscreen filters are the overview of P. Finkel in SOFW Journal 122, 543 (1996) refer to.

In addition to the two aforementioned groups of primary light stabilizers, it is also possible to use secondary light stabilizers of the antioxidant type which interrupt the photochemical reaction chain which is triggered when UV radiation penetrates into the skin. Typical examples thereof are amino acids (eg glycine, histidine, tyrosine, tryptophan) and their derivatives, imidazoles (eg urocanic acid) and their derivatives, peptides such as D, L-carnosine, D-carnosine, L-carnosine and their derivatives (eg anserine), chlorogenic acid and its derivatives, lipoic acid and its derivatives (eg dihydrolipoic acid), aurothioglucose, propylthiouracil and other thiols (eg thioredoxin, glutathione, cysteine, cystine, cystamine and their glycosyl , N-acetyl, methyl, ethyl, propyl, amyl, butyl and lauryl, palmitoyl, oleyl, γ-linoleyl, cholesteryl and glyceryl esters) and their salts, dilauryl thiodipropionate, distearyl thiodipropionate, Thiodipropionic acid and its derivatives (esters, ethers, peptides, lipids, nucleotides, nucleosides and salts) and sulfoximine compounds (eg buthionine sulfoximines, homocysteinesulfoximine, bithine sulfones, penta, hexa, heptathionine sulfoximine) in very low tolerated dosages (eg. pmol to μmol / kg), furthermore (Me tall) chelators (e.g. Α-hydroxy fatty acids, palmitic acid, phytic acid, lactoferrin), α-hydroxy acids (eg citric acid, lactic acid, malic acid), humic acid, bile acids, bile extracts, bilirubin, biliverdin, EDTA, EGTA and their derivatives, unsaturated fatty acids and their derivatives (eg γ-linolenic acid, linoleic acid, oleic acid), folic acid and its derivatives, ubiquinone and ubiquinol and their derivatives, vitamin C and derivatives (eg ascorbyl palmitate, Mg ascorbyl phosphate, ascorbyl acetate), tocopherols and derivatives (eg. Vitamin E acetate), vitamin A and derivatives (vitamin A palmitate), and benzylic resin, rutinic acid and derivatives thereof, α-glycosylrutin, ferulic acid, furfurylidene glucitol, carnosine, butylhydroxytoluene, butylhydroxyanisole, nordihydroguaiacetic acid, nordihydroguiaretic acid, trihydroxybutyrophenone, Uric acid and its derivatives, mannose and its derivatives, superoxide dismutase, zinc and its derivatives (eg ZnO, ZnSO ₄ ), selenium and its derivatives (eg S elen-methionine), stilbenes and their derivatives (e.g. B. stilbene oxide, trans-stilbene) and the present invention suitable derivatives (salts, esters, ethers, sugars, nucleotides, nucleosides, peptides and lipids) of these drugs.

In addition, compounds for suppressing or alleviating skin disorders induced by UV radiation, in particular activators of peroxisome proliferator-activated receptors (PPAR activators), can be added according to the invention, as in US Pat WO 02/38150 to which reference is hereby incorporated by reference.

• • Glycerin;
• • Alkylenglycole, wie beispielsweise Ethylenglycol, Diethylenglycol, Propylenglycol, Butylenglycol, Hexylenglycol sowie Polyethylenglycole mit einem durchschnittlichen Molekulargewicht von 100 bis 1.000 Dalton;
• • technische Oligoglyceringemische mit einem Eigenkondensationsgrad von 1,5 bis 10 wie etwa technische Diglyceringemische mit einem Diglyceringehalt von 40 bis 50 Gew.-%;
• • Methyolverbindungen, wie insbesondere Trimethylolethan, Trimethylolpropan, Trimethylolbutan, Pentaerythrit und Dipentaerythrit;
• • Niedrigalkylglucoside, insbesondere solche mit 1 bis 8 Kohlenstoffen im Alkylrest, wie beispielsweise Methyl- und Butylglucosid;
• • Zuckeralkohole mit 5 bis 12 Kohlenstoffatomen, wie beispielsweise Sorbit oder Mannit,
• • Zucker mit 5 bis 12 Kohlenstoffatomen, wie beispielsweise Glucose oder Saccharose;
• • Aminozucker, wie beispielsweise Glucamin;
• • Dialkoholamine, wie Diethanolamin oder 2-Amino-1,3-propandiol.

Hydrotropes such as, for example, ethanol, isopropyl alcohol, or polyols can also be used to improve the flow behavior. Polyols contemplated herein preferably have from 2 to 15 carbon atoms and at least two hydroxyl groups. The polyols may contain other functional groups, in particular amino groups, or be modified with nitrogen. Typical examples are

• • glycerin;
• Alkylene glycols such as ethylene glycol, diethylene glycol, propylene glycol, butylene glycol, hexylene glycol, and polyethylene glycols having an average molecular weight of 100 to 1,000 daltons;
• Technical Oligoglyceringemische with an intrinsic degree of condensation of 1.5 to 10 such as technical Diglyceringemische with a diglycerol content of 40 to 50 wt .-%;
• Methyolverbindungen, in particular trimethylolethane, trimethylolpropane, trimethylolbutane, pentaerythritol and dipentaerythritol;
• Lower alkyl glucosides, especially those having 1 to 8 carbons in the alkyl radical, such as, for example, methyl and butyl glucoside;
• Sugar alcohols having 5 to 12 carbon atoms, such as sorbitol or mannitol,
• • sugars having 5 to 12 carbon atoms, such as glucose or sucrose;
• • amino sugars, such as glucamine;
• Dialcohols, such as diethanolamine or 2-amino-1,3-propanediol.

When Preservatives are, for example, phenoxyethanol, Formaldehyde solution, parabens, pentanediol or sorbic acid and those listed in Appendix 6, Part A and B of the Cosmetics Regulation further substance classes.

Of the Total content of auxiliaries and additives may be 1 to 50, preferably 5 to 40 wt .-% - based on the means - amount. The preparation of the cosmetic and / or pharmaceutical preparation can be caused by usual cold or hot processes respectively; It is preferable to work according to the phase inversion temperature method.

The The following examples describe the invention, but without it limit:

example 1

CpG-1-PTO (5'-TCC ATG A CG TTC CTG A CG TT-3) and non-CpG-5 PTO (5'-CCC CCC CCC CCC CCC CCC CC-3 ') inhibit basal Akt activation , HaCaT keratinocytes were incubated with the mentioned DNA ODN for 30 minutes. Subsequently, the cells were lysed, the proteins extracted and separated by SDS-PAGE. In the Western blot, phosphorylations on serine 473 and threonine 308 were detected with phospho-specific antibodies. Uniform sample loading was checked with total-Akt antibodies.

Example 2:

Permanent inhibition of Akt by administration of 4 μM CpG-1 PTO (5'-TCC ATG A CG TTC CTG A CG TT-3) into HaCaT cells.

protein extraction and proof see example 1.

Example 3:

length-dependent Suppression of Akt by CpG-1 PTO deletion mutants. HaCaT cells were treated with 4 μM of the deletion mutants mentioned below CpG-1 PTO incubated for 30 minutes. In the picture were for clarity, the abbreviations listed below used for the oligonucleotides.

protein extraction and proof see example 1.

Example 4:

length-dependent Suppression of Akt by non-CpG-5 PTO deletion mutants. HaCaT Cells were treated with 4 μM of the deletion mutants listed below of non-CpG-5 PTO incubated for 30 minutes. In the picture for the sake of clarity, the ones listed below Abbreviations used for the oligonucleotides.

protein extraction and proof see example 1.

Example 5:

inhibition Act by natural DNA or DNA hasty products. HaCaT keratinocytes were added for 60 minutes below Incubated substances in the indicated concentrations.

• E. coli = Genomische DNA aus Escherichia coli
• C. perfr. = Genomische DNA aus Clostridium perfringens
• Salm. sp. = Genomische DNA aus Lachssperma
• Symb. = Symbioflor
• BCG = Bacillus Calmette Guerin
• CpG-1-PTO: 5'-TCC ATG ACG TTC CTG ACG TT-3

Protein extraction and detection see Example 1.

• E. coli = Genomic DNA from Escherichia coli
• C. perfr. Genomic DNA from Clostridium perfringens
• Salm. sp. = Genomic DNA from salmon sperm
• Symb. = Symbioflor
• BCG = Bacillus Calmette Guerin
• CpG-1 PTO: 5'-TCC ATG A CG TTC CTG A CG TT-3

Example 6:

Cell-species-specific effects of CpG-1-PTO (5'-TCC ATG A CG TTC CTG A CG TT-3) and non-CpG-5 PTO (5'-CCC CCC CCC CCC CCC CCC CC-3 ') Act. Different cell lines were incubated for 30 minutes with 4 μM each of CpG-1 PTO and non-CpG-5 PTO, respectively.

• A-431 (epidermoide Karzinomzelllinie)
• HaCaT (spontan immortalisierte Keratinozytenlinie)
• SZ (SZ95, Sebozytenlinie)
• Cos-7 (Nierenepithelzelllinie)
• Fib (primäre Hauffibroblasten)
• G361 (Melanomzelllinie)

Protein extraction and detection see Example 1.

• A-431 (epidermoid carcinoma cell line)
• HaCaT (spontaneously immortalized keratinocyte line)
• SZ (SZ95, sebocyte line)
• Cos-7 (kidney epithelial cell line)
• Fib (primary Hauffibroblasten)
• G361 (melanoma cell line)

Example 7:

• CpG-1-PTO: 5'-TCC ATG ACG TTC CTG ACG TT-3 non-CpG-5-PTO: 5'-CCC CCC CCC CCC CCC CCC CC-3'
  

CpG-1 PTO and non-CpG-5 PTO sensitize HaCaT keratinocytes for apoptosis by caspase activation. HaCaT keratinocytes were incubated for 24 h with ascending concentrations of staurosporine (STS, 0.1, 0.5, 1.0 μM). Similarly, the cells were additionally incubated with 4 μM each of CpG-1 PTO or non-CpG-5 PTO. Subsequently, the cells were lysed, the proteins extracted and separated by SDS-PAGE. In Western blot, the undigested and cleaved forms of caspase 8 and 3 were detected. The effect of staurosporine on the proteolytic activation of caspase 8 and 3 is enhanced in the presence of oligonucleotides. The left-hand diagram illustrates the involvement of caspases 8 and 3 in the context of apoptosis.

• CpG-1 PTO: 5'-TCC ATG A CG TTC CTG A CG TT-3 Non-CpG-5 PTO: 5'-CCC CCC CCC CCC CCC CC-3 '
  

Example 8:

CpG-1 PTO and non-CpG-5 PTO sensitize HaCaT keratinocytes to apoptosis by releasing cytochrome C. HaCaT keratinocytes were incubated for 24h with ascending concentrations of Stauro sporin (StS, 0.1, 0.5, 1.0 μM). Similarly, the cells were additionally incubated with 8 μM each of CpG-1 PTO or non-CpG-5 PTO. Subsequently, the cells were fractionated by mild lysis with digitonin. The content of cytochrome C in the cytosolic extract was determined in a commercial Cytochrome C immunoassay (R & D Systems, Wiesbaden, Germany).

Of the Effect of staurosporine on the cytosolic cytochrome content C is amplified in the presence of the oligonucleotides.

cytochrome C is the largest in vital cells Part in the mitochondria. Only in the context of apoptosis it comes to an accumulation of cytoplasmic cytochrome C. There acts Cytochrome C acts as an activator of caspase 9 and is thus a very early stage Apoptosis markers.

Example 9:

None Sensitization to apoptosis in sebocytes of the skin (SZ95) by CpG-1 PTO and non-CpG-5 PTO.

SZ95 Cells were incubated for 24h with ascending concentrations of staurosporine (STS, 0.1, 0.5, 1.0 μM). Analogous in addition, the cells were additionally each with 8 uM CpG-1-PTO or non-CpG-5-PTO incubated. Subsequently, the Lyses cells, extracts the proteins and separated by SDS-PAGE. Western blotting was the proteolytic activation of caspase 3, 8 and 9 demonstrated. There could be no increased effect of staurosporine in combination with the oligonucleotides Staurosporine alone can be shown.

Example 10:

None Sensitization to apoptosis in primary fibroblasts skin by CpG-1 PTO and non-CpG-5 PTO.

fibroblasts were used for 24h with ascending concentrations of staurosporine (STS, 0.1, 0.5, 1.0 μM). Similarly, the Cells additionally each with 8 uM CpG-1-PTO or incubated with non-CpG-5 PTO. Subsequently, the cells were lysed, the proteins are extracted and separated by SDS-PAGE. In the western The proteolytic activation of caspase 3 and 9 was detected. Caspase 8 was undetectable. It could not be reinforced Effect of staurosporine in combination with the oligonucleotides Staurosporine alone can be shown.

Example 11:

None Sensitization to apoptosis by CpG-9-PTO (= short deletion fragment of CpG-1-PTO) and nCpG-5G-PTO (= short deletion fragment of non-CpG-5-PTO) in HaCaT.

Cells were incubated for 24 h with ascending concentrations of staurosporine (STS, 0.1, 0.5, 1.0 μM). Similarly, the cells were additionally incubated with 8 μM each of CpG-1 PTO, CpG-9 PTO, non-CpG-5 PTO or nCpG-5G PTO. Subsequently, the cells were lysed, the proteins extracted and separated by SDS-PAGE. In Western blot, the proteolytic activation of caspase 8 was detected. CpG-1 PTO and n-CpG5 PTO as positive controls resulted in increased caspase degradation, CpG-9 PTO and nCpG-5G PTO had no effect.

Example 12:

non-CpG-5-PTO sensitizes HaCaT keratinocytes for apoptosis Release of cytochrome C - no effect of n-CpG-5G-PTO (= short deletion fragment non-CpG-5-PTO).

HaCaT Keratinocytes were incubated for 24h with ascending concentrations of staurosporine (StS, 0.1, 0.5, 1.0 μM). Analogous in addition, the cells were additionally each with 8 uM incubated with non-CpG-5 PTO or n-CpG-5G PTO.

Subsequently the cells were fractionated by mild lysis with digitonin.

Of the Content of cytochrome C in the cytosolic extract was in a commercial Cytochrome C Immunoassay (R & D Systems, Wiesbaden, Germany).

Of the Effect of staurosporine on the cytosolic cytochrome content C is amplified in the presence of non-CpG-5 PTO, the short fragment n-CpG-5G-PTO shows no effect.

cytochrome C is the largest in vital cells Part in the mitochondria. Only in the context of apoptosis it comes to an accumulation of cytoplasmic cytochrome C. There acts Cytochrome C acts as an activator of caspase 9 and is thus a very early stage Apoptosis markers.

Example 13:

• G3139-PTO: 5'-TCTCCCAGCGTGCGCCAT-3'
  

No sensitization to apoptosis by G3139 (Oblimersen, Genasense) in HaCaT and A431 keratinocytes. Cells were incubated for 24 h with ascending concentrations of G3139-PTO (2, 4, 8, 16, 32 μM). Subsequently, the cells were lysed, the proteins extracted and separated by SDS-PAGE. Caspases 3, 8 and 9 were detected in the Western blot. No proteolytic activation under the influence of G3139 PTO could be demonstrated. There was no caspase 8 immunoreactivity in A431 keratinocytes.

• G3139 PTO: 5'-TCTCCCAG CG TG CG CCAT-3 '
  

Example 14:

CpG-1-PTO as a representative of the CpG-B class, non-CpG-5-PTO and n-CpG-3-PTO super-induce concentration-dependent apoptosis in the presence of staurosporine.

HaCaT Cells were co-administered with staurosporine (Sts) (-, 0.01, 0.02, 0.05 μM) and ascending concentrations (1, 2, 4, 8, 16 μM) of (A) CpG-1-PTO, (B) non-CpG-5-PTO and (C) n-CpG-3-PTO treated. After 24 hours, the trial was terminated and the histone-associated DNA fragments in the cytosolic extract by means of a commercial Kits (Cell Death enzyme-linked immunosorbent assay, Boehringer Mannheim, Mannheim, Germany).

Example 15:

n-CpG-6-PTO inhibits and M-362-C (= CpG-C class) has no effect on the staurosporine-induced Apoptosis.

• n-CpG-6-PTO: 5'-GGG GGG GGG GGG GGG GGG GG-3' M-362-C: 5'-TCG tcg tcg ttc gaa cga cgt TGA T-3' (Großbuchstaben repräsentieren Phosphorothioat-Verknüpfungen, Kleinbuchstaben repräsentieren Phosphodiester-Verknüpfungen)
  

HaCaT cells were coadministered with staurosporine (Sts) (-, 0.01, 0.02, 0.05 μM) and ascending concentrations (1, 2, 4, 8, 16 μM) of (A) n-CpG-6. PTO and (B) M-362-C. After 24 h, the experiment was terminated and the histone-associated DNA fragments were determined in the cytosolic extract using a commercial kit (Cell Death enzyme-linked immunosorbent assay, Boehringer Mannheim, Mannheim, Germany).

• n-CpG-6 PTO: 5'-GGG GGG GGG GGG GGG GGG GG-3 ' M-362-C: 5'-TCG tcg tcg ttc gaa cga cgt TGA T-3 '(capital letters represent phosphorothioate linkages, lowercase letters represent phosphodiester linkages)
  

Example 16:

1585 (A) as a representative of the CpG class A shows a biphasic influence on staurosporine-induced apoptosis: small concentrations (1, 2 and 4 μM) have super-inducing, high concentrations (8 and 16 μM) have a suppressive effect.

• 1585(A): 5'-GGg gtc aac gtt gaG GGG Gg-3' (Großbuchstaben repräsentieren Phosphorothioat-Verknupfungen, Kleinbuchstaben repräsentieren Phosphodiester-Verknüpfungen)
  

HaCaT cells were treated concomitantly with staurosporine (Sts) (-, 0.01, 0.02, 0.05 μM) and ascending concentrations (1, 2, 4, 8, 16 μM) of 1585 (A). After 24 h, the experiment was terminated and the histone-associated DNA fragments were determined in the cytosolic extract using a commercial kit (Cell Death enzyme-linked immunosorbent assay, Boehringer Mannheim, Mannheim, Germany).

• 1585 (A): 5'-GGg gtc aac gtt gaG GGG Gg-3 '(capital letters represent phosphorothioate linkages, lowercase letters represent phosphodiester linkages)
  

It follows Sequence listing according to WIPO St. 25. This can of the official publication platform of the DPMA become.

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Cited patent literature

• - DE 10233994 A [0019]
• - DE 10361502 A [0024]
• - DE 19740092 A [0047]
• - DE 1165574 [0059]
• - DE 2024051 [0061]
• FR 2252840 A [0068]
• EP 0693471 B1 [0076]
• EP 0818450 A1 [0076]
• EP 0694521 B1 [0076]
• - DE 19712033 A1 [0078]
• WO 02/38150 [0080]

Cited non-patent literature

• Todd et al. in Cosm. Toil. 91, 27 (1976) [0070]
• - R. Lochhead in Cosm. Toil. 108, 95 (1993) [0075]
• P. Finkel in SOFW Journal 122, 543 (1996) [0078]

Claims (21)

Cosmetic and / or pharmaceutical preparation for the treatment of epithelial covering tissue, characterized in that it contains nucleic acids. Preparation according to claim 1, characterized in that that the treatment epithelial covering tissue in particular against hyperplastic disorders addressed epithelial covering tissue is. Preparation according to claim 1, characterized in that that the treatment epithelial covering tissue in particular on the Improvement of the appearance of the skin, the epilation or the induction of Hair loss is addressed and combined with cosmetic agents or UV light. Preparation according to one of the preceding claims, characterized in that they are used in combination with cosmetic Active ingredients or UV light as an anti-aging agent, in particular for the treatment of age spots, calluses or so-called Landmann's skin is used. Preparation according to one of the preceding claims, characterized in that they additionally at least an apoptosis-inducing substance, in particular 5-fluorouracil or imiquimod. Preparation according to one of the preceding claims, characterized in that the nucleic acids are selected are among synthetic nucleic acids, nucleic acids eukaryotic origin and nucleic acids bacterial Origin, in particular with nucleic acids from Escherichia coli and Clostridium perfringens. Preparation according to one of the preceding claims, characterized in that the nucleic acids are selected are among high molecular weight bacterial DNA, low molecular weight bacterial DNA, high molecular weight eukaryotic DNA, low molecular weight eukaryotic DNA and oligonucleotides. Preparation according to one of the preceding claims, characterized in that the oligonucleotides have a length from 5 to 100, especially 5 to 70, preferably 10 to 50, preferred 10 to 40, and most preferably from 12 to 30 nucleotides exhibit. Preparation according to one of the preceding claims, characterized in that the nucleic acids completely or partially chemically modified. Preparation according to claim 9, characterized that the chemical modification is selected under a) Change in the internucleoside bridges, in particular Exchange of phosphodiesters for methylphosphonates, phosphoramidates, Phosphorothioates or hydroxylamines; b) change the sugar components, in particular exchange of ribose for various Hexo- or pentopyranoses or 3'-5'-carbocyclic bridged Derivatives of 2'-deoxyribose; or c) replacement of the strand backbone, in particular replacement of polyester chains based on sugar-phosphate units against carboxamide chains based on amino acid derivatives, such as N- (2-aminoethyl) glycine units, where the a) mentioned under Replacement of phosphodiesters against phosphorothioates is particularly preferred is. Preparation according to claim 9 or 10, characterized that the nucleic acids phosphorothioate phosphodiester mixmere exhibit. Preparation according to one of the preceding claims, characterized in that they contain the nucleic acids in liposomes packed contains. Preparation according to one of the preceding claims, characterized in that the hyperplastic diseases of the epithelial cover tissue are selected from carcinogens and precancerous lesions and verrucae. Preparation according to one of the preceding claims, characterized in that the nucleic acids as oligonucleotides having one, two or more CpG motifs. Preparation according to one of the preceding claims, characterized in that the nucleic acid acids are present as oligonucleotides which have at least one CpG motif and belong to CpG-A or in particular to CpG-B class. Preparation according to one of the preceding claims, characterized in that the nucleic acids as a superstructure-forming Oligodeoxynukleotide present. Preparation according to Claim 16, characterized that the superstructure-forming nucleic acids are selected are among thymine-rich or cytosine-rich sequences with a Thymine or cytosine content of 25% to 100%, preferably 50% to 100%, more preferably 75% to 100%, and most preferably 100%. Preparation according to claim 17, characterized that the superstructure-forming nucleic acids are selected are among poly-T homopolymers or poly-C homopolymers. Use of nucleic acids as chemosensitizer for the treatment of hyperplasia epithelial covering tissue, for improvement of the skin, for the epilation or for the induction of hair loss. Use according to claim 19 in combination with cosmetic Active ingredients or UV light as an anti-aging agent, in particular for the treatment of age spots, calluses or so-called Landmann's skin. Use according to claim 19 or 20, characterized that in combination with other substances or forms of therapy takes place, in particular with those that are selected under: a) coal tar, b) phototherapy, c) photodynamic Therapy, preferably with 5-aminolevulinic acid, d) anthralin, e) Cryotherapy and f) Retinoids, preferably ATRA, acitretin and tazarotene